Conversion of Glycerol to 1,3-Propanediol

Microbial Conversion of Glycerol to 1,3-Propanediol: Physiological Comparison of a Natural Producer, Clostridium butyricum VPI 3266, and an Engineered Strain, Clostridium acetobutylicum DG1(pSPD5). Abstract: Clostridium acetobutylicum is not able to grow on glycerol as the sole carbon source since it cannot reoxidize the excess of NADH generated by glycerol catabolism. Nevertheless, when the pSPD5 plasmid, carrying the NADH-consuming 1,3-propanediol pathway from C. butyricum VPI 3266, was introduced into C. acetobutylicum DG1, growth on glycerol was achieved, and 1,3-propanediol was produced. In order to compare the physiological behavior of the recombinant C. acetobutylicum DG1(pSPD5) strain with that of the natural 1,3- propanediol producer C. butyricum VPI 3266, both strains were grown in chemostat cultures with glycerol as the sole carbon source. The same “global behavior” was observed for both strains: 1,3-propanediol was the main fermentation product, and the qH2 flux was very low. However, when looking at key intracellular enzyme evels, significant differences were observed. Firstly, the pathway for glycerol oxidation was different: C. butyricum uses a glycerol dehydrogenase and a dihydroxyacetone kinase, while C. acetobutylicum uses a glycerol kinase and a glycerol-3-phosphate dehydrogenase. Secondly, the electron flow is differentially regulated: (i) in C. butyricum VPI 3266, the in vitro hydrogenase activity is 10-fold lower than that in C. acetobutylicum G1(pSPD5), and (ii) while the ferredoxin-NAD
reductase activity is high and the NADH-ferredoxin reductase ctivity is low in C. acetobutylicum DG1(pSPD5), the reverse is observed for C. butyricum VPI 3266. Thirdly, actate dehydrogenase activity is only detected in the C. acetobutylicum DG1(pSPD5) culture, explaining why his microorganism produces lactate.

Production of 1,3-Propanediol from Glycerol by Clostridium cetobutylicum and Other Clostridium Species. Abstract: Glyceol was fermented with the production of 1,3-propanediol as the major fermentation product by four strains of Clostridium acetobutylicum, six of C. butylicum, two of C. beijerinckii, one of C. kainantoi, and three of C. butyricum. 1,3-Propanediol was identified by its retention times in gas chromatography and high-pressure liquid chromatography and by its mass spectrum. During growth of C. butylicum B593 in a chemostat culture at pH 6.5, 61% of the glycerol fermented was converted to 1,3-propanediol. When the pH was decreased to 4.9, growth and 1,3-propanediol production were substantially reduced.

Use of Glycerol from biodiesel production: Conversion to added value products. Abstract: A detailed study of main stages of a novel no-conventional process to obtain 1,3-propanediol (PD) from glycerol (Gly) has been made. Initially the volumetric productivity of Gly fermentation with Klebsiella pneumoniae bacterium was optimized for one and two continuous stages, where was necessary to study the multiplicity of stable steady states for the fermentation system selecting the conditions where higher concentrations of PD are found; and under optimal operation conditions the outlet PD concentration and the global yield are 0.4833 mol/l and 0.5481 molPD/molGly, respectively. For PD recovery from fermentation broth a no-conventional separation scheme was proposed, which consists first in a reactive-extraction with iso-butyraldehyde (iBAld) for produce 2-iso-propyl-1,3-dioxane (iPDOx) and water. The iPDOx is removed toward the organic phase by the aldehyde that acts simultaneously as reagent and solvent. The yield of reactive-extraction process is 85%. Finally Static Analysis (SA) for iPDOx hydrolysis system shows that is possible to obtain PD of high purity and recovery the aldehyde at the 88% in a reactive-distillation tower. SA also allowed obtaining the technological configuration of reactive-distillation tower and this technological synthesis was validated starting from simulations to both infinite/infinite and finite/finite (stages/reflux) conditions, using the ASPEN PLUS® software. For finite conditions the simulations showed that a conversion of 69% and 97.5% is reached for reflux ratios of 5.73 and 9.5 respectively.

HIGH PRODUCTION OF 1,3-PKOPANEDIOL FROM GLYCEROL BY Closhdium butyricum VPI 3266 IN A SIMPLY CONTROLLED FEDBATCH SYSTEM. Abstract: A simple fed-batch system which controls substrate feeding by measuring the CO, produced during the fermentation, was developped. This Fed-batch approach allowed high production of 1,3-propanediol from glycerol by Clostridium butyricum by avoiding substrate inhibition phenomena. 65 g/l of 1,3-propanediol was produced with a productivity of 1.21 g/l.h and a yield of 0.56. The concentration of 1,3-propanediol obtained and the productivity were significantly higher than those reached in batch culture.

CATALYTIC CONVERSION OF GLYCEROL AND SUGAR ALCOHOLS TO VALUE-ADDED PRODUCTS. Ph.D.Thesis.

Value-added Chemicals from Crude Glycerol

According to a new research paper, bBoth conventional catalysis and biological conversion can offer promising opportunities for the biodiesel industry to convert crude glycerol into a wide variety of value-added products.

A list of compounds that can be made via oxidation or reduction of glycerol are:

1. Tartronic acid (C3H3O5)
2. Dihydroxyacetone
3. Mesoxalic acid (Ketomalonic acid)
4. Glyceraldehydes
5. Glyceric acid
6. Malonic acid
7. Hydroxypyruvic acid
8. Lactic acid
9. Pyruvic acid
10. Propylene glycol
11. Propionic acid
12. Glycidol
13. Acrylic acid
14. Propanol
15. Isopropanol
16. Acetone
17. Propylene oxide
18. Propionaldehyde
19. Allyl alcohol
20. Acrolein
21. Hydrogen

Production of various coproducts via anaerobic fermentation of glycerol by clostridia:

1. 1,3-PDO
2. Acetic acid
3. Butyric acid
4. Lactic acid
5. Succinic acid
6. Butanol
7. Ethanol
8. Acetone

A new big cake for advanced biofuels: A real stimulus

DOE Announces Funding Opportunity of up to $200 Million for 5-12 Pilot and Demonstration Scale Biorefinery Projects. This is a real stimulus for the infant biofuels industry under the severe conditions of financial crisis, low oil price, and lack of confidence from people. A best Christmas gift for this industry.

The World's First Bio-based Succinic Acid Plant has been launched

According to BioBasedNews.com, A succinic acid plant from BioAmber with an annual production capacity of 2,000 metric tons has started construction in Pomacle Francein and expect to begin production in the fall of 2009.

The fate of glycerin from biodiesel production

Glycerin, or glycerine, is a by-product of biodiesel production. Supply of glycerin in the United States and worldwide is projected to grow over the next decade as government policies and incentives favor increased processing of plant oils for production of biodiesel fuels (For every 9 kilograms of biodiesel produced, about 1 kilogram of a crude glycerin by-product is formed; ). Sooner or later, the amount of glycerin produced will exceed the amount consumed by the market today. The prices in the glycerin market will continue to drop and eventually, glycerin will become an environmental liability.

Currently, up to 10% of the dietary dry matter could be supplied by glycerol with no decreases in feed intake or alterations of performance in growing ruminants or lactating cows. However,
there are some concerns with glycerin as a feedstuff due to its contents of methanol and mineral salts such as potassium salts and phosphates.

Today, biodiesel production plants are in need of methods to realize increased income from this glycerol and solve the fate of glycerol as a by-product.

One of the potentioal solutions is to convert the crude natural glycerol to propylene glycol. Some of the benefits of this technology are to utilize the waste and produce propylene glycol from a renewable source raw material. Of course, this technology could be used in biodiesel production plants to increase profitability.

Pervaporation–Membrane Process for Bioethanol Recovery

The traditional separation process, fractional distillation accounts for abou 40% of the cost for corn to ethanol production. A relatively new technology called "pervaporation" may provide considerable energy savings over traditional distillation technologies. Pervaporation is a membrane-based process used to separate and concentrate volatile compounds from a liquid mixture by selective permeation through a non-porous membrane into a vacuum permeate stream. The remaining liquid thus becomes depleted in the compound that permeates the membrane.

A US patent "Pervaporation process of separating a liquid mixture" disclosed an invention withe the abstract: " A pervaporation process for separating at least one component from a mixture of liquids, for example for separating ethanol from a fermentation mass, by a series of three separation steps: separating the mixture by a first pervaporation to form a first permeate vapor enriched in the component to be separated; fractionating the first permeate vapor, for example by temperature condensation, to form a high concentration fraction twice enriched in the component to be separated; and either distilling the high concentration fraction or a second pervaporation to form a distillate or retentate liquid thrice enriched in the component to be separated".

Other than carbon dioxide from bioethanol car

It is known that bioethanol is a greener, sustainable and renewable fuel which helps to reduce the greenhouse gases that contribute to global warming. A 10% ethanol blends can reduce greenhouse gas emissions by 12-19%. The carbon dioxide's provenance is a crucial factor in this reduction. Other emissions concerned are nitrogen oxides, particles, and hydrocarbons, which are also lower lowered when you run a car on bioethanol. Modern gasoline cars are already so clean that these improvements are of marginal importance. But the improvements are significant for diesel cars. The only measurable emission that increases with alcohols is aldehydes. With current catalysers, the level is so low that this is negligible.

Market demand drives cellulosic ethanol investment

The fate of cellulosic ethanol industry will be decided by current and future market demand. Jeff Broin, the CEO of Poet predicted, "Within 20 years, US ethanol producers will churn out 135 billion gallons of ethanol a year, two-thirds of which will be cellulosic ethanol. That would make the domestic ethanol industry the nation's largest source of liquid transportation fuel." Today, the annual US gasoline market is 140 billion gallons. Therefore, there is a great market demand potential, which are attracting continued investment in and development of future ethanol technology with the government's policy and financial support.

However, many investors are wondering how long the technology can be commercialized and when the cellulosic ethanol industry will be profitable?

Making biodiesel: easy or challenging?

From this website: "Anybody can make biodiesel. It's easy, you can make it in your kitchen.."This process is so-called conventional chemical catalysts production and widely used nowadays.

From this paper: "Enzymatic production of biodiesel has been proposed to overcome the drawbacks of the conventional chemically catalyzed processes. The main obstacle facing full exploitation of the enzyme, lipase, potential is its cost". Is it a direction to make biodiesel?

Waste Coffee Grounds Offer New Source Of Biodiesel Fuel

ScienceDaily (Dec. 15, 2008) — Researchers in Nevada are reporting that waste coffee grounds can provide a cheap, abundant, and environmentally friendly source of biodiesel fuel for powering cars and trucks.

A list of people and organizations working on microorganisms to convert C5 sugars to ethanol

Here is a list of people and organizations working on microorganisms to convert C5 sugars to ethanol:

1. Thomas W. Jeffries, Director of Institute for Microbial and Biochemical Technology, USDA
Microorganism: Pichia stipitis
Description of research: Xylose fermentation; Metabolic regulation; Metabolic engineering; Yeast genetics; Depolymerization of cellulosic and hemicellulosic polysaccharides; Lignin biodegradation; Bioprocess engineering; Microbial strain selection and development; Regulation of heterologous enzymes; Overproduction of primary and secondary metabolites; Overproduction of extracellular enzymes; Microbial physiology.

Novozymes used the strain provided by Thomas Jeffries to ferment the mixture of glucose and xylose.

2. Nancy W. Y. Ho, Research Molecular Biologist/Group Leader, Laboratory of Renewable Resources Engineering, Purdue University
Microorganism: Saccharomyces cerevisiae 259ST. A genetically modified version of 259A capable of xylose fermentation due to insertion of xylose reductase and xylitol dehydrogenase genes from P. stipitis and overexpression of xylulokinase.

Iogen used the engineered yeast developed by Dr. Ho to produce ethanol from wheat straw.UBC and Tembec Chemicals Products have tested 259ST yeast on fermenting spent sulfite pulping liquor (SSL).

3. Lonnie O. Ingram ,Distinguished Professor, Director, Florida Center for Renewable Chemicals and Fuels (FCRC),Department of Microbiology and Cell Science University of Florida.
Microorganism: Escherichia coli
General areas: Global redirection of central metabolism by genetic engineering; Industrial fermentation processes; Carbohydrate metabolism; Expression and secretion of glycohydrolases which degrade plant polymers; Alcohol tolerance

4. Lisbeth Olsson, Professor, DTU Biosys, Department of Systems Biology,Technical University of Denmark, Center for Microbial Biotechnology, BioCentrum-DTU
Microorganism: Saccharomyces cerevisiae strains (F12, CR4, and CB4)
Article abstract: Fermentations with three different xylose-utilizing recombinant Saccharomyces cerevisiae strains (F12, CR4, and CB4) were performed using two different wheat hemicellulose substrates, unfermented starch free fibers, and an industrial ethanol fermentation residue, vinasse. With CR4 and F12, the maximum ethanol concentrations obtained were 4.3 and 4 g/L, respectively, but F12 converted xylose 15% faster than CR4 during the first 24 h. The comparison of separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF) with F12 showed that the highest, maximum ethanol concentrations were obtained with SSF. In general, the volumetric ethanol productivity was initially, highest in the SHF, but the overall volumetric ethanol productivity ended up being maximal in the SSF, at 0.013 and 0.010 g/L.h, with starch free fibers and vinasse, respectively.

"Separate and Simultaneous enzymatic hydrolysis and fermentation of wheat hemicellulose with recombinant xylose utilizing Saccharomyces cerevisiae", Applied Biochemistry and Biotechnology, vol: 129-132, pages: 117-129, 2006
5. Taso, George, Laboratory of Renewable Resources Engineering, A. A. Potter Engineering Center, Purdue University
Microorganism: First using xylose isomerase and then yeast.
Article abstract:d-Xylulose, an intermediate of d-xylose catabolism, was observed to be fermentable to ethanol and carbon dioxide in a yield of greater than 80% by yeasts (including industrial bakers' yeast) under fermentative conditions. This conversion appears to be carried out by many types of yeast known for d-glucose fermentation. In some yeasts, xylitol, in addition to ethanol, was produced from d-xylulose. Fermenting yeasts are also able to produce ethanol from d-xylose when d-xylose isomerizing enzyme is present. The results indicate that ethanol could be produced from d-xylose in a yield of greater than 80% by a two-step process. First, d-xylose is converted to d-xylulose by xylose isomerase. d-Xylulose is then fermented to ethanol by yeasts.

"Production of Ethanol from d-Xylose by Using d-Xylose Isomerase and Yeasts", Appl Environ Microbiol. 1981 February; 41(2): 430–436.

6. Ronald Hector, Stephen Hughes and Xin Liang-Li, the research molecular biologists with the USDA's Agricultural Research Service.
Research: Optimizations of genes of commercial yeasts for production of fuel ethanol from hemicellulosic biomass are carried out to meet the rapidly expanding need for ethanol. These yeast strains will be used in cellulosic ethanol production.

7. Richard Bolin
US patent: 5372939: Combined enzyme: Schizosaccharoyces pombe, cellulase, .beta.-glucosidase, and xylose isomerase

8. Iogen, Recombinant yeasts (424A). The yeast strain was provided by Dr. Nancy Ho in Purdue University.

9. Xethanol, proceeding with a Cooperative Research and Development Agreeement (CRADA) with FPL to engineer genetically modified yeasts that will substantially increase fermentation time of such feedstocks such as xylose, to produce either ethanol or xylitol, a natural sweetener.

10. Lee R. Lynd, Chemical and Biochemical Engineering Program, Dartmouth College and CFO of Mascoma
Microorganism: Thermoanaerobacter thermosaccharolyticum HG-8.
Study the xylose-utilizing thermophiles, Thermoanaerobacterium saccharolyticum and Thermoanaerobacterium thermosaccharolyticum.

Mascoma’s research laboratories are now developing a new generation of microbes and processes for economical conversion of cellulosic feedstocks into ethanol.

11. Min Zhang, National Renewable Energy Laboratory, Applied Biological Sciences
Microorganism: Bacterium Zymomonas mobilis
The ethanol-producing bacterium Zymomonas mobilis was metabolically engineered to broaden its range of fermentable substrates to include the pentose sugar xylose. Two operons encoding xylose assimilation and pentose phosphate pathway enzymes were constructed and transformed into Z. mobilis in order to generate a strain that grew on xylose and efficiently fermented it to ethanol. This strain efficiently fermented both glucose and xylose, which is essential for economical conversion of lignocellulosic biomass to ethanol.
Metabolic Engineering of a Pentose Metabolism Pathway in Ethnologenic Z Mobilis. Science Vol. 267. 1995, pp. 240 - 243

12. VTT Technical Research Centre
Microorganism: Saccharomyces cerevisiae
They have studied recombinant Saccharomyces cerevisiae yeast metabolically engineered to utilise the pentose sugar xylose, and compared the gene expression profiles on xylose to glucose-metabolizing cells.

13. Jeffrey Tolan and R. K. Finn, School of Chemical Engineering, Cornell University
Microorganism: Erwinia chrysathemi
Fermentation of D-xylose and L-arabinose to ethanol by Erwinia chrysathemi, Appl. Envirom. Microbiol., 53(9): 2033-2038, 1987.

14. Alexander, M.A.; Chapman, T.W.;& Jeffries, T.W. Department of Chem. Eng. University of Wisconsin
Microorganism: Candida shehatae
Continuous xylose ferrmentation by Candida shehatae in a two-stage reactor. Applied biochemistry and biotechnology, v. 17:221-229, 1988

15. Satoshi Katahira1, Atsuko Mizuike, Hideki Fukuda1 and Akihiko Kondo, Department of Chemical Science and Engineering, Faculty of Engineering, Kobe University,
Microorganism: Recombinant yeast strain
Ethanol fermentation from lignocellulosic hydrolysate by a recombinant xylose- and cellooligosaccharide-assimilating yeast strain. Applied Microbiology and biotechnology, v. 72(6):1136-1143, 2006

16. Yong-su Jin, University of Illinois
Microorganism: E. coli
Improvement of xylose fermentation by recombinant Saccharomycescerevisiae through systematic and combinatorial approaches is funded by EBI (Energy Biosciences Institute). EBI is a new research organization that will pursue basic and applied research in the area of cellulosic biofuels, enhanced oil recovery, microbially-enhanced fossil fuel processing and biosequestration.

17. Girishchandra Patel, Division of Biological Sciences,National research council of Canada
Microorganism: Bacteroides polypragmatus
Fermentation of xylose and hemicellulose hydrolysates by an ethanol-adapted culture of Bacteroides polypragmatus, Archives of Microbiology, 146(1):68-73, 1986

Fuel Ethanol: Beyound Ethanol

1. Fuel ethanol is not sold with zero water content. Fuel ethanol with less than 0.5 % water is considered “anhydrous ethanol” while ethanol with higher water contents is usually referred to as “hydrated ethanol”.

2. Denatured alcohol typically contains up to 1% water and other constituents (e.g. methanol). Denaturing is to add denaturants to alcohol in order to make it unfit for beverage or internal human medical use.

3. A parameter that is used in evaluation of fuel ethanol is the pHe as defined by ASTM D6423. Like pH used to measure the acidity of aqueous solutions, the pHe value is a measure of the acid strength of high ethanol content fuels (>70% v/v). The pH 7 is considered neutral for aqueous solutions, whereas a pHe value of 9.55 is the neutralization point for ethanol.

4. Ethanolic solutions have lower conductivity than water.

5.The oxygen solubility in ethanol is higher than that of water.

A Possible Industrial Solution to Ferment Lignocellulosic Hydrolyzate to Ethanol

It is known that fermentable sugars can be obtained from lignocellulosic biomass through enzymatic or chemical hydrolysis processes. Although there are great interests in the application of enzymatic hydrolysis due to its specific products and high yield, in a short term, the time-consuming process and rather expensive enzyme cost make enzymatic hydrolysis still uncertain for use in large-scale industry.

One of the disadvantages of chemical hydrolysis process such as dilute-acid hydrolysis is the formation of by-products (furan derivatives, phenolic compounds, and aliphatic acids), which inhibit cultivation of yeast in lignocellulosic hydrolyzates. Hence on one hand, a great effort has to be made to reduce the inhibitor formation; on the other hand, detoxification step such as membrane separation is needed in the process. In addition, since the fermenting microorganisms are able to detoxify the hydrolyzates in situ, any process that can provide low concentration of the inhibitors and high cell density may be successful in cultivation of these toxic hydrolyzates. To some extent, continuous cultivation can be one of the approaches to provide low concentration of the inhibitors in the reactors and the high cell density can be provided by using cell recycling, immobilization or encapsulation.

According to s new research Paper, "a toxic dilute-acid hydrolyzate was continuously cultivated using a highcell-density flocculating yeast in a single and serial bioreactor which was equipped with a settler to recycle the cells back to the bioreactors. No prior detoxification was necessary to cultivate the hydrolyzates, as the flocks were able to detoxify it in situ. The experiments were successfully carried out at dilution rates up to 0.52 h-1. The cell concentration inside the bioreactors was between 23 and 35 g-DW/L, while the concentration in the effluent of the settlers was 0.32 ± 0.05 g-DW/L. An ethanol yield of 0.42-0.46 g/g-consumed sugar was achieved, and the residual sugar concentration was less than 6% of the initial fermentable sugar (glucose, galactose and mannose) of 35.2 g/L".

Bioethanol shippment

With more boethanol into the market, the issue of transporting ethanol will attract more attention.

In a short term, the shipments of ethanol to terminals can be handled mostly by tanker truck and rail tank car rather than pipelines. In the long term, can we ship the ethanol by pipeline as gasoline? If so, the following issues need to be aware of and solved:

1. Ethanol’s water affinity. Because of the high solubility of ethanol in water, water accumulation in pipelines is a normal occurrence, which may risks rendering ethanol unusable as a transportation fuel.
2. Corrosion. Ethanol-related corrosion problems can be caused ethanol behavior in the pipe. The typical one is stress corrosion cracking, which is very hard to detect. The main factors involve in the type of metal (carbon steel), physical environment (areas of stress concentration, near weld heat effect zone), chemical environment (Dissolved oxygen), and other minors. This damage may be accelerated at weld joints or “hard spots” where the steel metallurgy has been altered.

To address these issues, the technical feasibility is the first step; then significant investments in new and modified facilities and operational practices will be demanded.

Genomics strategies on biomass to biofuels

Based on the cover storty on C&EN, BESC's 300 scientists located at federal labs, academic institutions, companies, and nonprofit organizations across the country have been given the challenge of designing a path through the lignocellulosics recalcitrance problem.

Deconstructing lignocellulose to accessible sugars followed by chemical or fermentation processes is considered to be the most practical pathway to biofuels. However, pretreatment is a very expensive step in the process for biofuel production. So their efforts are to eliminate or significantly reduce the amount of pretreatment needed. One of the approaches is to ultimately come up with plants that are more easily digested and might need just a hot-water pretreatment without chemicals. A second approach is to engineer a multitalented microbe that can disassemble the plant cell wall and ferment the resulting sugars into biofuel in one go, a strategy known as consolidated bioprocessing. This type of one-organism, one-pot process could be a major breakthrough for low-cost production of ethanol or other fuels and chemicals.

BESC's approach to conquering recalcitrance has started with a major study on a fast-growing poplar tree from the Pacific Northwest and switchgrass native to the prairies of North America. Because no single parameter characterizes recalcitrance, the standard operating procedure is to screen multiple plant samples under a variety of conditions and measure the amount of sugar produced at the end of each test run.

Stimulus plan: Good news for "green" technology

Based on the report from The Wall Street Journal, green-technology advocates estimate that among President-elect Barack Obama’s $500 billion stimulus plan,the green component could be $50 billion or 10 % of the overall package for the complex green infrastructure initiatives -- such as building renewable energy plants, improving the electrical grid and installing "smart" meters that allow consumers to reap benefits from using electricity at off-peak hours. This would take effect well into the second year.

A collection of research papers on biobased succinic acid production

Modeling of batch fermentation kinetics for succinic acid production by Mannheimia succiniciproducens (2008). Kinetic models are proposed for the batch production of succinic acid from glucose by Mannheimia succiniciproducens MBEL55E. The models include terms accounting for both substrate and product inhibitions. Experimental data collected from a series of batch fermentations with different initial glucose concentrations were used to estimate parameters and also to validate the models proposed. The optimal values of the parameters were approximated by minimizing the discrepancy between the model predictions and corresponding experimental data. The growth of M. succiniciproducens could be expressed by a modified Monod model incorporating inhibitions of glucose and organic acids accumulated in the culture broth. The Luedeking–Piret model was able to describe the formation of organic acids as the fermentation proceeded, in which succinic, acetic, and formic acids followed a mixed-growth-associated pattern. However, unexpectedly, lactic acid fermentation by M. succiniciproducens was nearly nongrowth-associated. In all cases, the model simulation matched well with the experimental observations, which made it possible to elucidate the fermentation characteristics of M. succiniciproducens during efficient succinic acid production from glucose. These models thus can be employed for the development and optimization of biobased succinic acid production processes.

Succinic Acid Production by Continuous Fermentation Process Using Mannheimia succiniciproducens LPK7 (2008). To achieve a higher succinic acid productivity and evaluate the industrial applicability, this study used Mannheimia succiniciproducens LPK7 (knock-out: ldhA, pflB, pta-ackA), which was recently designed to enhance the productivity of succinic acid and reduce by-product secretion. Anaerobic continuous fermentation of Mannheimia succiniciproducens LPK7 was carried out at different glucose feed concentrations and dilution rates. After extensive fermentation experiments, a succinic acid yield and productivity of 0.38 mol/mol and 1.77 g/l/h, respectively, were achieved with glucose feed concentration of 18.0 g/l and 0.2 h-1 dilution rate. A similar amount of succinic acid production was also produced in batch culture experiments. Therefore, these optimal conditions can be industrially applied for the continuous production of succinic acid. To examine the quantitative balance of the metabolism, a flux distribution analysis was also performed using the metabolic network model of glycolysis and the pentose phosphate pathway.

From genome sequence to integrated bioprocess for succinic acid production by Mannheimia succiniciproducens (2008). Mannheimia succiniciproducens is a capnophilic gram-negative bacterium isolated from bovine rumen.Wild-type M. succiniciproducens can produce succinic acid as a major fermentation product with acetic, formic, and lactic acids as byproducts during the anaerobic cultivation using several different carbon sources. Succinic acid is an important C4 building block chemical for many applications. Here, we review the progress made with M. succiniciproducens for efficient succinic acid production; the approaches taken towards the development of an integrated process for succinic acid production are described, which include strain isolation and characterization, complete genome sequencing and annotation, development
of genetic tools for metabolic engineering, strain development by systems approach of integrating omics and in silico metabolic analysis, and development of fermentation and recovery processes. We also describe our current effort on further improving the performance of M. succiniciproducens and optimizing the mid- and downstream processes. Finally, we finish this mini-review by discussing the issues that need to be addressed to make this process of fermentative succinic acid production employing M. succiniciproducens to reach the industrial-scale process.

Prospects for a bio-based succinate industry. Bio-based succinate is receiving increasing attention as a potential intermediary feedstock for replacing a large petrochemical-based bulk chemical market. The prospective economical and environmental benefits of a bio-based succinate industry have motivated research and development of succinate-producing organisms. Bio-based succinate is still faced with the challenge of becoming cost competitive against petrochemical-based alternatives. High succinate concentrations must be produced at high rates, with little or no byproducts to most efficiently use substrates and to simplify purification procedures. Herein are described the current prospects for a bio-based succinate industry, with emphasis on specific bacteria that show the greatest promise for industrial succinate production. The succinate-producing characteristics and the metabolic pathway used by each bacterial species are described, and the advantages and disadvantages of each bacterial system are discussed.

Succinic acid production using metabolically engineered Escherichia coli (2007). Thesis.

Simultaneous fermentation and crystallization in succinic acid (2006). MS Thesis.

Production of Succinic Acid by E.coli from Mixtures of Glucose and Fructose (2005), a thesis by Andreas Lennartsson evaluated whether AFP184 can utilize fructose, both alone and in mixtures with glucose, as a carbon source for the production of succinic acid. The results showed that the Escherichia coli strain AFP184 can utilize fructose both alone and in mixtures with glucose. A succinic acid concentration of 52 g/L was reached with a mixture of fructose and glucose. The corresponding mass yield was 0.71 gram succinic acid per gram anaerobically consumed sugar. It was also shown that a high initial concentration of glucose (100 g/L) did not yield high levels of acetate during fermentations with Escherichia coli strain AFP184.

Fermentative Production of Succinic Acid from Glucose and Corn Steep Liquor by Anaerobiospirillum succiniciproducens (2005), a research by by Lee et al found that A. succiniciproducens was able to grow in a minimal medium containing glucose when supplemented with corn steep liquor (CSL) as the sole complex nitrogen source. The concentration of CSL had a significant effect on the glucose consumption by A. succiniciproducens. When 10-15 g/L of CSL was supplemented, cells were grown to an OD660 of 3.5 and produced 17.8 g/L succinic acid with 20 g/L glucose. These results are similar to those obtained by supplementing yeast extract and polypeptone, thereby suggesting that succinic acid can be produced more economically using glucose and CSL.

In Silico Metabolic Pathway Analysis and Design: Succinic Acid Production by Metabolically Engineered Escherichia coli as an Example (2002). In this study, Lee et al have constructed in silico metabolic pathway network of Escherichia coli consisting of 301 reactions and 294 metabolites. Metabolic ux analyses were carried out to estimate ux distributions to achieve the maximum in silico yield of succinic acid in E. coli. The maximum in silico yield of succinic acid was only 83% of its theoretical yield. The lower in silico yield of succinic acid was found to be due to the insufficient reducing power, which could be increased to its theoretical yield by supplying more reducing power. Furthermore, the optimal metabolic pathways for the production of succinic acid could be proposed based on the results of metabolic flux analyses. In the case of succinic acid production, it was found that pyruvate carboxylation pathway should be used rather than phosphoenolpyruvate carboxylation pathway for its optimal production in E. coli. Then, the in silico optimal succinic acid pathway was compared with conventional succinic acid pathway through minimum set of wet experiments.The results of wet experiments indicate that the pathway predicted by in silico analysis is more efficient than conventional pathway.

Kinetic study for the extraction of succinic acid with TOA in Fermentation broth; effects of pH, salt and contaminated acid. Reactive extraction can be used for the recovery of carboxylic acids from fermentation broth. Through the formation of complex with extractants at the two-phase interface, the carboxylic acids are partitioned into organic solvents. However, the recovery of carboxylic acids is interrupted by the conditions of fermentation broth. In this study, the effects of conditions of fermentation broth on the extraction kinetics were investigated using a microporous membrane-based stirred cell for the extraction of succinic acid with tri-n-octylamine. The interfacial concentrations of species in various systems were correlated and thus the effects of pH, salts and contaminated acid on the intrinsic reaction kinetics were discovered. The reaction rate constants were determined from the forward reaction rate equation reported in our previous work. It was found that the extraction rates were steeply decreased at pH values larger than 3 due to the dissociation of carboxylic group. Competitive extraction of salts, and contaminated acid, which was pyruvic acid, had negative influence on the extraction process of succinic acid and thus the extraction rates were decreased. The interfacial concentrations of succinic acid and TOA in fermentation broth had no difference with those in artificial single acid systems. Therefore, the decrease of extraction rates can be explained by the change of ionic strength in fermentation broth.

Isolation and characterization of succinic-acid producing Mannheimia sp. from bovine rumen. A novel succinic acid-producing bacterium was isolated from bovine rumen. The bacterium is a non-motile, non-spore-forming, mesophilic and capnophilic gram-negative rod or coccobacillus. Phylogenetic analysis based on the 16S rRNA sequence and physiological analysis indicated that the strain belongs to the recently reclassified genus Mannheimia as a novel species, and has been named Mannheimia succiniciproducens MBEL55E. Under 100% CO2 conditions, it grows well in the pH range of 6.0–7.5 and produces succinic acid, acetic acid and formic acid at a constant ratio of 2:1:1. When M. succiniciproducens MBEL55E was cultured anaerobically in medium containing 20 g l–1 glucose as carbon source, 13.5 g l–1 of succinic acid was produced.

Succinic Acid Adsorption from Fermentation Broth and Regeneration (2004). More than 25 sorbents were tested for uptake of succinic acid from aqueous solutions. The best resins were then tested for successive loading and regeneration using hot water. The key desired properties for an ideal sorbent are high capacity, complete stable regenerability, and specificity for the product. The best resins have a stable capacity of about 0.06 g of succinic acid/g of resin at moderate concentrations (1–5 g/L) of succinic acid. Several sorbents were tested more exhaustively for uptake of succinic acid and for successive loading and regeneration using hot water. One resin, XUS 40285, has a good stable isotherm capacity, prefers succinate over glucose, and has good capacities at both acidic and neutral pH. Succinic acid was removed from simulated media containing salts, succinic acid, acetic acid, and sugar using a packed column of sorbent resin, XUS 40285. The fermentation byproduct, acetate, was completely separated from succinate. A simple hot water regeneration successfully concentrated succinate from 10 g/L (inlet) to 40–110 g/L in the effluent. If successful, this would lower separation costs by reducing the need for chemicals for the initial purification step. Despite promising initial results of good capacity (0.06 g of succinic/g of sorbent), 70% recovery using hot water, and a recovered concentration of >100 g/L, this regeneration was not stable over 10 cycles in the column. Alternative regeneration schemes using acid and base were examined. Two (XUS 40285 and XFS-40422) showed both good stable capacities for succinic acid over 10 cycles and >95% recovery in a batch operation using a modified extraction procedure combining acid and hot water washes. These resins showed comparable results with actual broth.

Effective purification of succinic acid from fermentation broth produced by Mannheimia succiniciproducens (2006). The present study deals with the development of purification and separation processes required to produce the highly purified succinic acid from the fermentation broth produced by recombinant microorganism, Mannheimia succiniciproducens. The newly developed process consists of the pretreatment process such as reactive extraction and vacuum distillation step and the crystallization process for the highly purified succinic acid production. By-produced acids were effectively removed by the reactive extraction as a primary separation. In addition, the crystallization was applied without adding any salts to produce highly purified succinic acid. The purified succinic acid, with 99.8% purity and 73.1% yield rate was obtained through this newly developed purification process.

Improved Succinic Acid Production in the Anaerobic Culture of an Escherichia coli pflB ldhA Double Mutant as a Result of Enhanced Anaplerotic Activities in the Preceding Aerobic Culture (2007). Escherichia coli NZN111 is a pflB ldhA double mutant which loses its ability to ferment glucose anaerobically due to redox imbalance. In this study, two-stage culture of NZN111 was carried out for succinic acid production. It was found that when NZN111 was aerobically cultured on acetate, it regained the ability to ferment glucose with succinic acid as the major product in subsequent anaerobic culture. In two-stage culture carried out in flasks, succinic acid was produced at a level of 11.26 g/liter from 13.4 g/liter of glucose with a succinic acid yield of 1.28 mol/mol glucose and a productivity of 1.13 g/liter _ h in the anaerobic stage. Analyses of key enzyme activities revealed that the activities of isocitrate lyase, malate dehydrogenase, malic enzyme, and phosphoenolpyruvate (PEP) carboxykinase were greatly enhanced while those of pyruvate kinase and PEP carboxylase were reduced in the acetate-grown cells. The two-stage culture was also performed in a 5-liter fermentor without separating the acetate-grown NZN111 cells from spent medium. The overall yield and concentration of succinic acid reached 1.13 mol/mol glucose and 28.2 g/liter, respectively, but the productivity of succinic acid in the anaerobic stage dropped to 0.7 g/liter _ h due to cell autolysis and reduced anaplerotic activities. The results indicate the great potential to take advantage of cellular regulation mechanisms for improvement of succinic acid production by a metabolically engineered E. coli strain.

Genome-Based Metabolic Engineering of Mannheimia succiniciproducens for Succinic Acid Production (2006). Succinic acid is a four-carbon dicarboxylic acid produced as one of the fermentation products of anaerobic metabolism. Based on the complete genome sequence of a capnophilic succinic acid-producing rumen bacterium, Mannheimia succiniciproducens, gene knockout studies were carried out to understand its anaerobic fermentative metabolism and consequently to develop a metabolically engineered strain capable of producing succinic acid without by-product formation. Among three different CO2-fixing metabolic reactions catalyzed by phosphoenolpyruvate (PEP) carboxykinase, PEP carboxylase, and malic enzyme, PEP carboxykinase was the most important for the anaerobic growth of M. succiniciproducens and succinic acid production. Oxaloacetate formed by carboxylation of PEP was found to be converted to succinic acid by three sequential reactions catalyzed by malate dehydrogenase, fumarase, and fumarate reductase. Major metabolic pathways leading to by-product formation were successfully removed by disrupting the ldhA, pflB, pta, and ackA genes. This metabolically engineered LPK7 strain was able to produce 13.4 g/liter of succinic acid from 20 g/liter glucose with little or no formation of acetic, formic, and lactic acids, resulting in a succinic acid yield of 0.97 mol succinic acid per mol glucose. Fed-batch culture of M. succiniciproducens LPK7 with intermittent glucose feeding allowed the production of 52.4 g/liter of succinic acid, with a succinic acid yield of 1.16 mol succinic acid per mol glucose and a succinic acid productivity of 1.8 g/liter/h, which should be useful for industrial production of succinic acid.

Metabolic Engineering of Escherichia coli for Enhanced Production of Succinic Acid, Based on Genome Comparison and In Silico Gene Knockout Simulation (2005). Comparative analysis of the genomes of mixed-acid-fermenting Escherichia coli and succinic acid-overproducing Mannheimia succiniciproducens was carried out to identify candidate genes to be manipulated for overproducing succinic acid in E. coli. This resulted in the identification of five genes or operons, including ptsG, pykF, sdhA, mqo, and aceBA, which may drive metabolic fluxes away from succinic acid formation in the central metabolic pathway of E. coli. However, combinatorial disruption of these rationally selected genes did not allow enhanced succinic acid production in E. coli. Therefore, in silico metabolic analysis based on linear programming was carried out to evaluate the correlation between the maximum biomass and succinic acid production for various combinatorial knockout strains. This in silico analysis predicted that disrupting the genes for three pyruvate forming enzymes, ptsG, pykF, and pykA, allows enhanced succinic acid production. Indeed, this triple mutation increased the succinic acid production by more than sevenfold and the ratio of succinic acid to fermentation products by ninefold. It could be concluded that reducing the metabolic flux to pyruvate is crucial to achieve efficient succinic acid production in E. coli. These results suggest that the comparative genome analysis combined with in silico metabolic analysis can be an efficient way of developing strategies for strain improvement.

Succinic acid production and purification (US patent, 1999). A highly efficient process for the production and recovery of pure succinic acid from a succinate salt that involves minimal use of additional reagents, and produces virtually no waste by-products, and permits internal recycle of the base and acid values, is provided. The method involves the formation of diammonium succinate, either by using an ammonium ion based material to maintain neutral8 pH in the fermenter or by substituting the ammonium cation for the cation of the succinate salt created in the fermenter. The diammonium succinate can then be reacted with a sulfate ion, such as by combining the diammonium succinate with ammonium bisulfate and/or sulfuric acid at sufficiently low pH to yield succinic acid and ammonium sulfate. The ammonium sulfate is advantageously cracked thermally into ammonia and ammonium bisulfate. The succinic acid can be purified with a methanol dissolution step. Various filtration, reflux and reutilization steps can also be employed.

Fermentation and purification process for succinic acid (US Patent, 1992). A process for economically producing highly purified succinic acid comprises growing a succinate-producing microorganism on a low cost carbohydrate substrate; simultaneously neutralizing the fermentation broth and precipitating the succinate as calcium succinate by adding a calcium ion source to form calcium succinate; isolating the calcium succinate; slurrying the calcium succinate in water and treating it with sulfuric acid to form calcium sulfate and succinic acid; and then treating the succinic acid with first a strongly acidic ion exchanger and then a weakly basic ion exchanger to remove impurities and obtain a highly purified succinic acid product. In a preferred embodiment, the calcium succinate is isolated from the fermentation broth by filtration; the filtrate is heated to precipitate additional calcium succinate; and, the spent filtrate which contains nutrients is recycled to the fermentor.

Batch and continuous cultures of Mannheimia succiniciproducens MBEL55E for the production of succinic acid from whey and corn steep liquor (2003). Mannheimia succiniciproducens MBEL55E isolated from bovine rumen is able to produce a large amount of succinic acid in a medium containing glucose, peptone, and yeast extract. In order to reduce the cost of the medium, whey and corn steep liquor (CSL) were used as substrates for the production of succinic acid by M. succiniciproducens MBEL55E. Anaerobic batch cultures of M. succiniciproducens MBEL55E in a whey-based medium containing CSL resulted in the production of succinic acid with a yield of 71% and productivity of 1.18 g/l/h, which are similar to those obtained in a whey-based medium containing yeast extract (72% and 1.21 g/l/h). Anaerobic continuous culture of M. succiniciproducens MBEL55E in a wheybased medium containing CSL resulted in a succinic acid yield of 69% and a succinic acid productivity as high as 3.90 g/l/h. These results show that succinic acid can be produced efficiently and economically by M. succiniciproducens MBEL55E from whey and CSL.

Influence of C02-HC03- Levels and pH on Growth,Succinate Production, and Enzyme Activities of Anaerobiospirillum succiniciproducens (1992). Growth and succinate versus lactate production from glucose by Anaerobiospirillum succiniciproducens was regulated by the level of available carbon dioxide and culture pH. At pH 7.2, the generation time was almost doubled and extensive amounts of lactate were formed in comparison with growth at pH 6.2. The succinate yield and the yield of ATP per mole of glucose were significantly enhanced under excess-CO2-HCO3 growth conditions and suggest that there exists a threshold level of CO2 for enhanced succinate production in A. succiniciproducens. Glucose was metabolized via the Embden-Meyerhof-Parnas route, and phosphoenolpyruvate carboxykinase levels increased while lactate dehydrogenase and alcohol dehydrogenase levels decreased under excess-CO2-HC03- growth conditions. Kinetic analysis of succinate and lactate formation in continuous culture indicated that the growth rate-linked production rate coefficient (K) cells was much higher for succinate (7.2 versus 1.0 g/g of cells per h) while the non-growth-rate-related formation rate coefficient (K') was higher for lactate (1.1 versus 0.3 g/g of cells per h). The data indicate that A. succiniciproducens, unlike other succinate-producing anaerobes which also form propionate, can grow rapidly and form high final yields of succinate at pH 6.2 and with excess CO2-HCO3 as a consequence of regulating electron sink metabolism.

A list of companies for bioethanol and specialty biochemical production

If you are interested in bioethanol and specialty biochemicals, you canot ingnore these companies:

1. The companies on bioethanol and specialty biochemical production

Abengoa Bioenergy, a subsidiary of Abengoa S.A,has its headquarters in St. Louis, Missouri. Regarding their activity front, the company is opening a pilot facility for the cellulosic ethanol in York, Nebraska.

ADM is working with the abundant and renewable products of agriculture to develop nature-based alternatives to the world’s finite stores of fossil fuels.

American Process Inc is prominent in the cellulosic biorefinery field. Our proprietary process, AVAP™ (American Value Added Pulping) co-produces pulp and ethanol from wood in an integrated biorefinery application.

ARKENOL Fuels is a California (USA)-based technology and project development company whose focus is the construction and operation of ethanol factories worldwide. The company also licenses its ethanol-producing technology to others.

BioEnergy International, LLC is a privately-held, science and technology company leading in the development and commercialization of next generation biorefineries for the production of high-value bio-based chemicals and fuels from renewable feedstock through the use of its proprietary biocatalyst technology.

BioPetrol is developing a commercially viable global solution for turning sewage sludge into oil, saving enormous sums in disposal and saving the earth from the destructive sewage dumping which is ruining the environment.

BlueFire Ethanol Inc use the Concentrated Acid Hydrolysis patented process positions it as the viable, world-wide cellulose-to-ethanol company with demonstrated production experience with ethanol from wood wastes, urban trash (post-sorted MSW), rice and wheat straws and other agricultural residues.

BP is forming partnerships with academia and joint ventures with businesses to develop advanced technologies to boost the use of biofuels.

ButylFuel,LLC (BFL) will produce and market butanol as a solvent, with the future intent of selling butanol as a fuel.

Cargill has been adding value to things that grow for 140 years. Today Caegill apply that expertise to renewable fuels.

Catalyst Renewables Corp’s objective is to acquire and develop a portfolio of renewable power generation projects in the U.S. To-date the company has focused exclusively on baseload renewable power projects. Advances in technology and renewable portfolio standards implemented in numerous states have created an environment where renewable power projects can be developed profitably.

Changing World Technologies, Inc. (CWT) is at the forefront of revolutionizing renewable energy by changing the way in which organic waste materials are utilized, thereby providing a platform for sustainable development. What was once deemed as “waste” and transported to a landfill can now be converted to a reliable stream of renewable energy, minimizing global warming, and improving our quality of life.CWT’s subsidiaries and affiliate companies include:Resource Recovery Corporation, Inc.(RRC),Thermo-Depolymerization Process, LLC (TDP, LLC) , and Renewable Environmental Solutions, LLC (RES).

Chevron Technology Ventures identifies, develops and commercializes emerging technologies that have the potential to transform energy production and use. The business development portfolio includes: Biofuels ,Hydrogen infrastructure ,Emerging energy applications, and Venture capital.

Chief Energy has been providing Clean, Safe Oil Heat since 1977. The company is family owned and operates in the NY City metropolitan area including the 5 boroughs, Nassau & Westchester Counties.

China Resources Alcohol Corporation (CRAC) is the second largest ethanol producer in China and the owner of the cellulosic ethanol pilot demonstration plant in the world which operates continuously, 24-hours per day

CHOREN is one of the world’s leading gasification technology companies for solid biomass and oil based residue feedstock. The center-piece of the technology is the patented Carbo-V process that made the production of tar-free synthetic combustion gas possible and provided the breakthrough for the conversion of biomass to energy.

Cleantech Partners is a Wisconsin-based, private, non-profit organization that invests in emerging, energy-saving technologies.

CleanTech Biofuels is a development stage company with technology that the company believes is capable of converting municipal solid waste into ethanol and other products. By using the existing infrastructure for municipal solid waste collection and disposal to collect biomass at low or possibly negative feedstock cost, the Company expects to achieve profitability quickly relative to other energy producers who must develop their infrastructure to collect and transport more expensive feedstocks such as sugar cane, corn or even switchgrass, wood waste, or corn stover.

ClearFuels Technology Inc mission is simply stated in its name, to produce clear clean renewable fuels such as ethanol, methanol, hydrogen and synthetic gas from sustainable cellulosic biomass using advanced thermochemical technologies. ClearFuels focus is on the sugarcane platform, the source of over half the world’s ethanol. Clearfuels Technology is integrating its highly efficient thermochemical production of cellulosic ethanol from bagasse and cane trash with the established fermentation processes of producing ethanol from sugarcane.

Colusa Biomass Energy Corporation is a biomass-to-energy company focusing on bio-fuels for transportation.

Coskata is a biology-based renewable energy company for the low-cost production of ethanol from a wide variety of input material including biomass, municipal solid waste and other carbonaceous material.

DuPont Biofuels will provide advantaged products for agricultural energy crops, feedstock processing and advanced biofuels.

Dyadic International is engaged in the development, manufacture and sale of biological products using a number of proprietary fungal strains to produce enzymes and other biomaterials, principally focused on a system for protein production based on the patented Chrysosporium lucknowense fungus, known as C1. Dyadic uses, for itself and others, its patented and proprietary technologies to conduct research and development activities for the discovery, development, and manufacture of products and enabling solutions to the bioenergy, industrial enzyme and pharmaceutical industries.

Dynamotive Energy Systems Corporation is an energy solutions provider headquartered in Vancouver, Canada, with offices in the USA and Argentina. Its carbon/ greenhouse gas neutral fast pyrolysis technology uses medium temperatures and oxygen-free conditions to turn dry waste biomass and energy crops into BioOil® for power and heat generation. BioOil® can be further converted into vehicle fuels and chemicals. Dynamotive’s process does not require feedstock that has alternative food use.

EnerGenetics Energies, LLC (EGE) is one of the oldest biofuels research companies in the U.S. (est. 1978). EGE approaches the production of biofuels in a fundamentally different way than the traditional methods employed in current ethanol/bio-diesel operations. EGE believes that the key to economic sustainability in the biofuels industry is first to remove petroleum from the production of biofuels so that petroleum prices are de-linked from bio-fuel production costs, and second to produce value added products that recover feedstock costs associated with bio-fuel production. After 30 years of research and development, EGE has developed MBR technologies in conjunction with the USDA and leading universities which accomplishes these goals.

Energy Quest, Inc and its wholly owned subsidiary Syngas Energy Corp. (SEC) are active in the research, development and commercialization of “alternative energy” technologies. The term alternative encompasses green and/or renewable energies.

Ensyn is the world leader in the production of bio-oil produced from pyrolysis of renewable feedstocks. Ensyn was incorporated in 1984 to commercialize its proprietary biomass to liquid technology, Rapid Thermal Process (RTP)™. Ensyn currently provides the world’s rapid pyrolysis process that has operated on a long-term commercial basis.

Fuel Frontiers Inc’socus is on developing multiple ethanol synthesis facilities throughout the world. The facilities will be the first U.S. ethanol plant to employ plasma gasification technology in conjunction with the catalytic production of alcohol fuels from synthesis gas to transform an environmental problem (scrap tires and other wastes) into an earth-friendly ethanol solution.

Global Energy Holdings Group is focused on the markets in alternative and renewable energies.
The subsidiary Global Energy Systems is a provider of outsourced renewable and alternative energy projects and services. GES is working to transform alternative feed stocks into electricity, natural gas or liquid fuels.The subsidiary Global Energy Ventures invests in cutting edge companies in the clean technology industry.

Green Biologics Limited (GBL) focuses on fermentation-based biomass conversion technologies for the production of fuels and chemicals.

GreenField Ethanol Inc began in 1989 as Commercial Alcohols and has grown to be Canada's leading fuel and packaged alcohol producer. The company was rebranded in 2006 as GreenField Ethanol to better reflect the growing importance of its fuel ethanol business.

Green Energy Live is a company in the emerging waste/biomass-to-ethanol industry that converts wastes that are currently being landfilled, into ethanol and other valuable co-products using the proprietary patented gasification and conversion technology.

Green Star Products, Inc. (GSPI) is an environmentally friendly public company involved in the production of renewable clean burning fuels such as biodiesel and cellulosic ethanol and other products including super-lubricants that are designed to reduce emissions and improve fuel economy in vehicles, machinery and power plants.

GreenShift develops and commercializes technologies that facilitate the efficient use of natural resources. It does so today by developing and integrating new clean technologies into existing biofuel production facilities, by selling equipment based on our technologies, and by using our technologies to directly produce and sell biomass-derived oils and fuels.

KL Energy specializes in bio-fuels project development, engineering, construction, and plant management with an emphasis on ethanol made from both cellulose and grain.

Iogen Corporation is a world leading biotechnology firm specializing in cellulosic ethanol - a fully renewable, advanced biofuel that can be used in today's cars. Iogen also develops, manufactures and markets enzymes used to modify and improve the processing of natural fibres within the textile, animal feed, and pulp and paper industries.

Lignol is a Canadian company based in BC which is undertaking to construct biorefineries for the production of fuel-grade ethanol and biochemicals from Canadian forests and vast supplies of biomass feedstocks. Lignol has acquired and since modified, a solvent based pre-treatment technology that was originally developed by a subsidiary of General Electric. The pre-treatment technology was previously commercialized in a major pulp mill by Repap with an investment of over $C 100 million. Lignol recently established a Cellulosic Ethanol Development Centre in Vancouver which consists of a pilot plant, a state of the art enzyme development laboratory and an engineering group.

LS9, Inc., the Renewable Petroleum Company™, is a privately-held industrial biotechnology company based in South San Francisco, California developing patent-pending biofuels made with the power of synthetic biology. LS9 DesignerBiofuels™ products are customized to closely resemble petroleum fuels, engineered to be clean, renewable, domestically produced, and cost competitive with crude oil.

Masada Resource Group, LLC (Masada) is a provider of waste disposal services to address today's environmental issues. Through the development of its patented CES OxyNol™ process, Masada is the business of processing and converting municipal solid waste (MSW) and sewer sludge to fuel ethanol and other commercial byproducts. The environmentally safe, commercial CES OxyNol™ process transforms MSW, waste water and sewer sludge into valuable renewable energy sources.

Mascoma is pursuing the development of advanced cellulosic ethanol technologies across a range of cellulosic feedstocks. As part of our strategy of technology discovery, development and deployment, Mascoma is patenting numerous technologies and forming a broad set of research and commercial partnerships.

Mississippi Ethanol LLC (ME) is a privately owned, small business specializing in the development of renewable alternative energy through conversion of biomass. ME has worked to perfect the conversion of biomass through gasification to synthesis gas for power and liquid fuels production. ME is presently operating both an engineering-scale facility (3 ton/day) at Mississippi State University and a 30 ton/day production facility in Winona. The patented gasification system is a low-to-medium pressure, medium temperature (non-slagging) design with no air or oxygen addition for reaction. The gasification technology processing scheme uses simple, familiar unit operation systems with emphasis placed on ease of control and process safety.

New Generation Biofuels is a development stage renewable fuels provider. The company holds an exclusive license for North America, Central America and the Caribbean to commercialize proprietary technology to manufacture alternative biofuels from vegetable oils and animal fats that we intend to market as a new class of biofuel for power generation, heavy equipment, and marine use and as heating fuel.

Poet, the largest dry mill ethanol producer in the United States, is an established company in the bio-refining industry through project development, design and construction, research and development, plant management, and marketing. Formerly known as Broin, the 20-year old company markets more than one billion gallons of ethanol annually, has built 25 ethanol production facilities in the United States, 19 of which it currently operates, and has six more under construction or in development.

PureVision Technology, Inc. (“PureVision”) has developed and patented technologies that hold the promise of making the cellulosic biorefining industry technologically successful and profitable. PureVision’s carbon-neutral biomass fractionation technology converts abundant cellulosic biomass into sugars, energy and fiber that are bio-based raw materials to make many industrial and consumer products.

Qteros (formerly SunEthanol) is dedicated to producing low-carbon fuel energy from plant and tree waste.

Range Fuels is a privately held company funded primarily in the U.S. focusing on alternative, clean (green) energy systems. Our leadership team melds experience from the technologically intensive oil, chemical, petrochemical, coal gasification, power and gas-to-liquids industries, the renewable fuel industry, and the pulp and paper industry.

Shell Global Solutions provides business and operational consultancy, technical services and research and development expertise to the energy and processing industries worldwide.

SunOpta BioProcess Inc. (formerly Stake Technology Ltd.,) was founded in 1973 and is the original division of the SunOpta organization, specializes in the design, construction and optimization of biomass conversion equipment and facilities.

Syntec Biofuel Inc is a renewable energy company that is developing and commercializing proprietary second-generation biofuel technology and processes to convert waste cellulosic biomass into ethanol and other high-value alcohols. The company’s Biomass-to-Alcohols (B2A) thermo-chemical process can utilize virtually any organic feedstock, such as woodchips, corn stover, sugar bagasse, wheat straw and so forth, to produce highly sustainable and renewable fuel.

Tamarack Energy has focused on biomass-derived electric power generation as a successful strategy to address the rapidly expanding market for renewable energy. This focus reflects the market opportunities associated with renewable energy generated using biomass.

Universal Bioenergy is a Mississippi-based company engaged in the production of renewable fuels through its subsidiary Universal Bioenergy North America, Inc. who operates a biodiesel refinery in Mississippi. The refinery intends to produce biodiesel fuel from various virgin vegetable oils, premium greases, and non-edible vegetable oil sources using their unique and economical process.

Virent was founded in 2002 to commercialize the aqueous phase reforming (APR) process, which generates hydrogen from sugar, as originally published in the journal Nature. Researchers at Virent have since further advanced the technology into the BioForming® process, which combines APR with other catalytic technologies to produce renewable liquid fuels, fuel gases, and other chemicals.

Verenium Corporation, merger of Diversa and Celunol, a company in the development and the commercialization of cellulosic ethanol, an environmentally-friendly and renewable transportation fuel, as well as higher performance specialty enzymes for applications within the biofuels, industrial, and animal nutrition and health markets.

Virgin Green Fund has been established to invest in companies in the renewable energy and resource efficiency sectors in the US and Europe. It is a sector-focused, multi-stage investment firm investing primarily in expansion/growth capital opportunities with an allocation to earlier stage venture capital opportunities.

W2 Energy is a green energy company whose primary business is the production of liquid fuels (diesel, gasoline, methanol, butanol) from bio-mass, waste and coal feedstock.

Waste Management is North America's largest operator of LFGTE facilities, with renewable energy projects at 112 of its landfills. Upon completion of the 60-project expansion begun in 2007, Waste Management expects to generate over 700 megawatts of energy from its landfills.

Worldwide BioEnergy is an LLC formed for the technical advancement and commercialization of waste products into energy and other profitable by-products. WorldWide BioEnergy also is pursuing other promising technologies that will have a positive impact on the supply of green energy through conversion of waste into useful products. Worldwide BioEnergy approach is to insure the technology is feasible, develop a standardized conversion system that is economically practical and to partner with researchers as well as investor groups to take the technology to the market place.

ZeroPoint Clean Tech, Inc is a renewable energy technology and project development company. ZeroPoint has developed a highly efficient biomass gasification process capable of converting biomass into renewable synthesis gas, electricity, or liquid fuels (Cellulosic Diesel™, ethanol, or methanol).


2. The Companies for Enzyme-Developing, Genetic-Engineering, and Plant-Breeding

Agrivida is an agricultural biotechnology company developing energy crops designed to produce chemicals, fuels, and bioproducts from non-food cellulosic biomass.

Amyris Biotechnologies uses synthetic biology techniques to create new metabolic pathways in industrial microbes to produce novel or rare chemicals. Its primary project to date has focused on the use of synthetic biology to address supply and cost constraints limiting the use of the anti-malarial drug artemisinin.

BASF – the Chemical Company – consolidated all its plant biotechnology activities in BASF Plant Science in 1998. With subsidiaries such as CropDesign and metanomics, BASF Plant Science forms the industry’s leading research and technology platform.

BICAL is a company in the production and continued development of Miscanthus, the multipurpose crop for energy and industry.

Bioenergy International

Ceres is developing the crops needed for a new generation of biofuels and biopower.

Codexis is a clean technology company and develops biocatalysts used to create efficient and cleaner chemistry-based manufacturing processes in the life sciences, bioindustrial and chemical marketplaces. Codexis technology is used by global pharmaceutical companies for cost-effective manufacturing of human therapeutics and in the energy industry to enable advanced biofuels.

DuPont

Dyadic International, Inc is a global biotechnology company with the technology that brings nature to the marketplace. Dyadic is focused on the discovery, development, and manufacturing of novel products derived from the DNA of complex living organisms - including humans - found in the earth’s biodiversity.

Edenspace seeks to transform the energy, agricultural and environmental industries through innovative applications of plants for renewable fuels and environmental sustainability.

Farmacule is developing molecular farming technology to cost effectively mass produce high-value industrial and therapeutic proteins and biofuels.

Genencor is a leading industrial biotechnology company that develops and markets innovative enzymes and bio-based products.

Genentech has been delivering on the promise of biotechnology for more than 30 years, using human genetic information to discover, develop, manufacture and commercialize biotherapeutics that address significant unmet medical needs.

Lucigen Corporation focuses on developing new, much more effective products and technologies for gene cloning and genomics. Our patented CloneSmart® technology dramatically improved DNA cloning reliability and efficiency, allowing successful cloning of genes that had been impossible to clone with standard methods.

Metabolix brings sustainable, clean solutions to the world in plastics, energy, and chemicals by combining bioscience and nature.

Mendel is a plant biotechnology company that develops products with enhanced yield and quality focused on row crops and cellulosic biofeedstocks.

Monsanto is an agricultural company to apply innovation and technology to help farmers around the world produce more while conserving more.

Novozymes is the multinational biotech company.

Plant Biofuels Corporation Sdn Bhd (PBC) is a producer of renewable and environment friendly fuel, BIODIESEL.

Syngenta is a world-leading agri-business committed to sustainable agriculture through innovative research & technology.

Synthetic Genomics Inc is developing novel genomic-driven strategies to address global energy and environmental challenges. Recent advances in the field of synthetic genomics present seemingly limitless applications that could revolutionize production of energy, chemicals and pharmaceuticals and enable carbon sequestration and environmental remediation.

Verenium Corporation

Clean energy are key to Obama stimulus plan

Here is the voice from Obama during his campain: “Green jobs are the jobs of the future, not just because they pay well and can’t be outsourced…and not just because they’ll help strengthen our economy and lift up our middle class. But because they’ll help reduce our dependence on foreign oil, and save this planet for our children.”

Recent news from Los Anglos Times: "President-elect Barack Obama is crafting a financial stimulus plan far more ambitious than a quick cash infusion, viewing it as a way to create jobs in a struggling economy and push the nation toward improved healthcare and roads and clean energy."

It is a good news for alternative energy industry. The question is: this is an emergying industry and there is still long way to go to become a sector that can generate a lot of jobs in a short term. Before that, more money still need to be input to support its survival. Hopefully the time to reach that stage will be shortened due to the government support from policy and finance.

Downstream processing of biobased succinic acid (5)

Approach V: Sorption-Based SA Purification
(Davison et al. Appl. Biochem. Biotech, Vol. 113-116,653 (2004))

Key points for the process
–Key desirable properties for an ideal sorbent
--High capacity: >0.06 g SA/ g resin at moderate concentrations (1-5 g/l)
--Complete low-cost stable regenerability: 70% recovery using hot water and >95% using a modified extraction combining acid and hot washes
--Specificity for the product: a recovered concentration> 100 g/L

What about the feasibility for commercialization?

A step forward for butanol

As reported, a collaboration between Energy Research Institute of Shandong Academy of Sciences (SDERI), China and Green Biologics Ltd. (GBL), UK is established for Bio-butanol Production in China. It is one more step forward for butanol production.

Downstream processing of biobased succinic acid (4)

Approach IV: Reactive Extraction-Based SA Purification: Extraction and Crystallization (Huh, et al. Process Bio. 41(2006)1461-1465

Key points for the process

• TOA has high extractability and used for selective removal of contaminated organic acids and salts from dilute fermentation broth
  --Only extract the undissociated form of carboxylic acids
  -- Need multistage extraction using different degrees of dissociation of each acid with pH


• Vacuum distillation
  --Concentrate the pretreated broth and remove volatile organic acids such as acetic acid, formic acid etc.


• Crystallization
  --The crystal of SA is formed more than other organic acids in saturated fermentation broth (after vacuum distillation) and low solubility

Any comments for this method? Advantages and disadvantages? Cost or simplexity?

NACD president warns of further regulations

NACD president warns of further regulations. The changes occurring in Washington, D.C., now are sure to impact chemical distribution and transportation.

Transgenic plants containing ligninase and cellulase which degrade lignin and cellulose to fermentable sugars

This patent discloses transgenic plants and a method of producing such plants which degrade lignins and celluloses into fermentable sugars. The plants comprise ligninase and cellulase genes isolated from Acidothermus cellulolyticus, Thermomonospora fusca etc. The process includes, A) providing a first and second transgenic plant which includes a DNA encoding a cellulose and ligninase respectively operably linked to a nucleotide sequence encoding a signal peptide and B) mating by sexual fertilization the first and the second transgenic plants to produce a third transgenic plant which includes both the first and second DNA encoding the cellulase and ligninase, wherein the transgenic plant degrades the lignocellulose when ground to produce the plant material.

Process for the production of liquid fuels from biomass

US patent: Process for the production of liquid fuels from biomass

This patent discloses a process for continuously producing a pulp biomass, comprising subjecting a biomass containing feed to a treatment which comprises bringing the feed to a pressure of 100-250 bar, keeping the pressurized feed subsequently or concomitantly at a temperature not exceeding 280 C over a period of up to 60 minutes, thereby obtaining a pulp and optionally subjecting the pulp to a reaction step in which the pulp is heated over a. period of up to 60 minutes to a temperature exceeding 280C, resulting in the continuous production of a hydrocarbon product having a greater energy density than biomass.

It looks like this process use a little lower temperature than the traditional thermochemical conversion of biomass into liquid fuel.

Increased Ethanol Production from Xylose

US Patent: Increased Ethanol Production from Xylose

This invention provides methods of manipulating the carbon flux of a host cell transformed with plasmids of the invention. Plasmids of the invention may include nucleotides that encode pyruvate decarboxylase. In one embodiment, a strain of the thermotolerant yeast Hansenula polymorpha that has been transformed with plasmids and polynucleotides of the invention is provided.

Novozymes Inaugurates World's Largest Enzyme Fermentation Facility in China

Novozymes announced the inauguration of its Suzhou Hongda Enzymes Company production facility in Taicang, China, about 30 miles north of Shanghai. The company said the new plant , the largest enzyme fermentation facility in the world, would focus on products for the bioethanol industry. Novozymes executive vice president Peder Holk Nielsen said, "We believe bioethanol is a good example how biotechnology can make more from less, decoupling economic growth from the use of natural resources. As the world leader in bioinnovation, Novozymes is optimistic about the future of bioethanol and is dedicated to increasing our capabilities continuously in this field. The Suzhou facility is one of Novozymes' strategic manufacturing locations, and this new expansion will enable us to accomplish more."According to the press release, "Novozymes offers a highly efficient technology platform for sustainable production for bioethanol - getting more out of fewer resources. In 2010 Novozymes will have enzymes available on a large scale for production of second-generation bioethanol based on agricultural byproducts."

Despite the recent downturn in the global economy, Novozymes has increased its investment in Taicang, China, generating more revenue and jobs there. What attracts Novozyme invest more in China?

The news said , the United States, Brazil, and China have defined clear targets and roadmaps for the development of the bioethanol industry. By 2010 China aims to more than double its bioethanol production to cover 5% of the total transport fuel used with a target of 3 million tons fuel ethanol.

Why not in the USA?