Pilot-Scale Production of Fatty Acid Ethyl Esters by an Engineered Escherichia coli Strain Harboring the p(Microdiesel) Plasmid

ABSTRACT Fatty acid ethyl esters (FAEEs) were produced in this study by the use of an engineered Escherichia coli p(Microdiesel) strain. Four fed-batch pilot scale cultivations were carried out by first using glycerol as sole carbon source for biomass production before glucose and oleic acid were added as carbon sources. Cultivations yielded a cell density of up to 61 ± 3.1 g of cell dry mass (CDM) per liter and a maximal FAEE content of 25.4% ± 1.1% (wt/wt) of CDM.

[1]  F. Srienc,et al.  Trace: Tennessee Research and Creative Exchange Metabolic Engineering of Escherichia Coli for Efficient Conversion of Glycerol into Ethanol , 2022 .

[2]  J. Kelly,et al.  The two alcohol dehydrogenases of Zymomonas mobilis. Purification by differential dye ligand chromatography, molecular characterisation and physiological roles. , 1986, European journal of biochemistry.

[3]  Rainer Kalscheuer,et al.  Microdiesel: Escherichia coli engineered for fuel production. , 2006, Microbiology.

[4]  J. Knappe,et al.  Ultrastructure and pyruvate formate-lyase radical quenching property of the multienzymic AdhE protein of Escherichia coli. , 1992, The Journal of biological chemistry.

[5]  L. Ingram,et al.  Genetic engineering of ethanol production in Escherichia coli , 1987, Applied and environmental microbiology.

[6]  T. Voelker,et al.  Modification of the substrate specificity of an acyl-acyl carrier protein thioesterase by protein engineering. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[7]  E. Lustbader,et al.  Inheritability of plasmids and population dynamics of cultured cells. , 1975, Proceedings of the National Academy of Sciences of the United States of America.

[8]  L. Wackett Microbial‐based motor fuels: science and technology , 2008, Microbial biotechnology.

[9]  G. Strobel,et al.  The production of myco-diesel hydrocarbons and their derivatives by the endophytic fungus Gliocladium roseum (NRRL 50072). , 2008, Microbiology.

[10]  A. Steinbüchel,et al.  In vitro and in vivo biosynthesis of wax diesters by an unspecific bifunctional wax ester synthase/acyl‐CoA:diacylglycerol acyltransferase from Acinetobacter calcoaceticus ADP1 , 2003 .

[11]  J. Cronan,et al.  Overproduction of Acetyl-CoA Carboxylase Activity Increases the Rate of Fatty Acid Biosynthesis in Escherichia coli * , 2000, The Journal of Biological Chemistry.

[12]  C. Wills,et al.  Characterization of the two alcohol dehydrogenases of Zymomonas mobilis. , 1981, Archives of biochemistry and biophysics.

[13]  A. Steinbüchel,et al.  Formation of intracytoplasmic lipid inclusions by Rhodococcus opacus strain PD630 , 1996, Archives of Microbiology.

[14]  A. Steinbüchel,et al.  The Wax Ester Synthase/Acyl Coenzyme A:Diacylglycerol Acyltransferase from Acinetobacter sp. Strain ADP1: Characterization of a Novel Type of Acyltransferase , 2005, Journal of bacteriology.

[15]  A. Steinbüchel,et al.  Accumulation of storage lipids in species of Rhodococcus and Nocardia and effect of inhibitors and polyethylene glycol , 1997 .

[16]  H. Alvarez Relationship between β-oxidation pathway and the hydrocarbon-degrading profile in actinomycetes bacteria , 2003 .

[17]  C. Ratledge,et al.  The biochemistry and molecular biology of lipid accumulation in oleaginous microorganisms. , 2002, Advances in applied microbiology.

[18]  J. Keasling,et al.  Microbial production of fatty-acid-derived fuels and chemicals from plant biomass , 2010, Nature.

[19]  D. Peters,et al.  Carbohydrates for fermentation , 2006, Biotechnology journal.

[20]  L. Ingram,et al.  Construction and expression of an ethanol production operon in Gram-positive bacteria. , 2005, Microbiology.

[21]  Seraphim Papanikolaou,et al.  Lipid production by Yarrowia lipolytica growing on industrial glycerol in a single-stage continuous culture. , 2002, Bioresource technology.

[22]  Hauke Harms,et al.  Debaryomyces hansenii — an extremophilic yeast with biotechnological potential , 2006, Yeast.

[23]  Rhk Jinhe Acetone Butanol Fermentation of Rice Straw \nzymatic Hydrolysate , 1998 .

[24]  A. Steinbüchel,et al.  Lipid storage compounds in marine bacteria , 1997, Applied Microbiology and Biotechnology.

[25]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[26]  N. M. Packter,et al.  Importance of stored triacylglycerols in Streptomyces: possible carbon source for antibiotics. , 1994, Microbiology.

[27]  L. Ingram,et al.  Efficient ethanol production from glucose, lactose, and xylose by recombinant Escherichia coli , 1989, Applied and environmental microbiology.

[28]  Hwa Sung Shin,et al.  Optimal fed-batch operation of recombinant cells subject to plasmid instability and death , 2008, Bioprocess and biosystems engineering.

[29]  A. Steinbüchel,et al.  Establishment of a novel anabolism-based addiction system with an artificially introduced mevalonate pathway: complete stabilization of plasmids as universal application in white biotechnology. , 2009, Metabolic engineering.

[30]  Rainer Kalscheuer,et al.  Synthesis of Novel Lipids in Saccharomyces cerevisiae by Heterologous Expression of an Unspecific Bacterial Acyltransferase , 2004, Applied and Environmental Microbiology.

[31]  Alexander Steinbüchel,et al.  Fatty acid alkyl esters: perspectives for production of alternative biofuels , 2010, Applied Microbiology and Biotechnology.

[32]  M. K. Gouda,et al.  Single cell oil production by Gordonia sp. DG using agro-industrial wastes , 2008 .

[33]  A. Corma,et al.  Synthesis of transportation fuels from biomass: chemistry, catalysts, and engineering. , 2006, Chemical reviews.

[34]  U. Rinas,et al.  Simple fed-batch technique for high cell density cultivation of Escherichia coli. , 1995, Journal of biotechnology.

[35]  V. Fried,et al.  Organic Solvents as Probes for the Structure and Function of the Bacterial Membrane: Effects of Ethanol on the Wild Type and an Ethanol-Resistant Mutant of Escherichia coli K-12 , 1973, Journal of bacteriology.

[36]  K. Zhao,et al.  Microbial Conversion of Glycerol to 1,3-Propanediol by an Engineered Strain of Escherichia coli , 2009, Applied and Environmental Microbiology.

[37]  J. Nielsen,et al.  Fuel ethanol production from lignocellulose: a challenge for metabolic engineering and process integration , 2001, Applied Microbiology and Biotechnology.

[38]  Y. Nakamura,et al.  Growth inhibitory and lethal effects of ethanol on Escherichia coli , 1987, Biotechnology and bioengineering.

[39]  Lian Pin Koh,et al.  Potential habitat and biodiversity losses from intensified biodiesel feedstock production. , 2007, Conservation biology : the journal of the Society for Conservation Biology.

[40]  A. Steinbüchel,et al.  Thio Wax Ester Biosynthesis Utilizing the Unspecific Bifunctional Wax Ester Synthase/Acyl Coenzyme A:Diacylglycerol Acyltransferase of Acinetobacter sp. Strain ADP1 , 2005, Applied and Environmental Microbiology.

[41]  Xuefeng Lu,et al.  Overproduction of free fatty acids in E. coli: implications for biodiesel production. , 2008, Metabolic engineering.

[42]  Gerhard Knothe History of Vegetable Oil-Based Diesel Fuels , 2010 .

[43]  T. W. Jeffries,et al.  Bacteria engineered for fuel ethanol production: current status , 2003, Applied Microbiology and Biotechnology.

[44]  A. Steinbüchel,et al.  A Novel Bifunctional Wax Ester Synthase/Acyl-CoA:Diacylglycerol Acyltransferase Mediates Wax Ester and Triacylglycerol Biosynthesis inAcinetobacter calcoaceticus ADP1* , 2003, The Journal of Biological Chemistry.

[45]  J. Guinea,et al.  Cellular lipid accumulation by Pseudomonas aeruginosa 44T1 , 1991, Applied Microbiology and Biotechnology.

[46]  T. Voelker,et al.  Alteration of the specificity and regulation of fatty acid synthesis of Escherichia coli by expression of a plant medium-chain acyl-acyl carrier protein thioesterase , 1994, Journal of bacteriology.

[47]  Jürgen Krahl,et al.  The Biodiesel Handbook , 2005 .

[48]  P. Cunningham,et al.  Cloning and sequence analysis of the fermentative alcohol-dehydrogenase-encoding gene of Escherichia coli. , 1989, Gene.