Production of Odd-Carbon Dicarboxylic Acids in Escherichia coli Using an Engineered Biotin–Fatty Acid Biosynthetic Pathway

2017 ◽  
Vol 139 (13) ◽  
pp. 4615-4618 ◽  
Author(s):  
Robert W. Haushalter ◽  
Ryan M. Phelan ◽  
Kristina M. Hoh ◽  
Cindy Su ◽  
George Wang ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Fatty Acid ◽  
Biosynthetic Pathway ◽  
Dicarboxylic Acids ◽  
Fatty Acid Biosynthetic Pathway

scholarly journals Engineering Escherichia coli FAB system using synthetic plant genes for the production of long chain fatty acids

2019 ◽  
Vol 18 (1) ◽  
Author(s):  
Elias Kassab ◽  
Monika Fuchs ◽  
Martina Haack ◽  
Norbert Mehlmer ◽  
Thomas B. Brueck
Keyword(s):  
Escherichia Coli ◽  
Fatty Acids ◽  
Fatty Acid ◽  
Production System ◽  
Biosynthetic Pathway ◽  
Long Chain Fatty Acids ◽  
E Coli ◽  
Fatty Acid Biosynthetic Pathway ◽  
Long Chain ◽  
Chain Fatty Acids

Abstract Background Sustainable production of microbial fatty acids derivatives has the potential to replace petroleum based equivalents in the chemical, cosmetic and pharmaceutical industry. Most fatty acid sources for production oleochemicals are currently plant derived. However, utilization of these crops are associated with land use change and food competition. Microbial oils could be an alternative source of fatty acids, which circumvents the issue with agricultural competition. Results In this study, we generated a chimeric microbial production system that features aspects of both prokaryotic and eukaryotic fatty acid biosynthetic pathways targeted towards the generation of long chain fatty acids. We redirected the type-II fatty acid biosynthetic pathway of Escherichia coli BL21 (DE3) strain by incorporating two homologues of the beta-ketoacyl-[acyl carrier protein] synthase I and II from the chloroplastic fatty acid biosynthetic pathway of Arabidopsis thaliana. The microbial clones harboring the heterologous pathway yielded 292 mg/g and 220 mg/g DCW for KAS I and KAS II harboring plasmids respectively. Surprisingly, beta-ketoacyl synthases KASI/II isolated from A. thaliana showed compatibility with the FAB pathway in E. coli. Conclusion The efficiency of the heterologous plant enzymes supersedes the overexpression of the native enzyme in the E. coli production system, which leads to cell death in fabF overexpression and fabB deletion mutants. The utilization of our plasmid based system would allow generation of plant like fatty acids in E. coli and their subsequent chemical or enzymatic conversion to high end oleochemical products.

scholarly journals Identification and Characterization of a New Enoyl Coenzyme A Hydratase Involved in Biosynthesis of Medium-Chain-Length Polyhydroxyalkanoates in Recombinant Escherichia coli

2003 ◽  
Vol 185 (18) ◽  
pp. 5391-5397 ◽  
Author(s):  
Si Jae Park ◽  
Sang Yup Lee
Keyword(s):  
Escherichia Coli ◽  
Fatty Acid ◽  
Chain Length ◽  
Biosynthetic Pathway ◽  
Pha Synthase ◽  
Enzymatic Analysis ◽  
E Coli ◽  
Medium Chain ◽  
Medium Chain Length ◽  
Enoyl Coa Hydratase

ABSTRACT The biosynthetic pathway of medium-chain-length (MCL) polyhydroxyalkanoates (PHAs) from fatty acids has been established in fadB mutant Escherichia coli strain by expressing the MCL-PHA synthase gene. However, the enzymes that are responsible for the generation of (R)-3-hydroxyacyl coenzyme A (R3HA-CoAs), the substrates for PHA synthase, have not been thoroughly elucidated. Escherichia coli MaoC, which is homologous to Pseudomonas aeruginosa (R)-specific enoyl-CoA hydratase (PhaJ1), was identified and found to be important for PHA biosynthesis in a fadB mutant E. coli strain. When the MCL-PHA synthase gene was introduced, the fadB maoC double-mutant E. coli WB108, which is a derivative of E. coli W3110, accumulated 43% less amount of MCL-PHA from fatty acid compared with the fadB mutant E. coli WB101. The PHA biosynthetic capacity could be restored by plasmid-based expression of the maoCEc gene in E. coli WB108. Also, E. coli W3110 possessing fully functional β-oxidation pathway could produce MCL-PHA from fatty acid by the coexpression of the maoCEc gene and the MCL-PHA synthase gene. For the enzymatic analysis, MaoC fused with His6-Tag at its C-terminal was expressed in E. coli and purified. Enzymatic analysis of tagged MaoC showed that MaoC has enoyl-CoA hydratase activity toward crotonyl-CoA. These results suggest that MaoC is a new enoyl-CoA hydratase involved in supplying (R)-3-hydroxyacyl-CoA from the β-oxidation pathway to PHA biosynthetic pathway in the fadB mutant E. coli strain.

A Cellular Assay for Inhibitors of the Fatty Acid Biosynthetic Pathway Using Scintillating Microplates

2015 ◽  
Vol 13 (5) ◽  
pp. 285-292 ◽  
Author(s):  
Thao Ung ◽  
Jennifer L. Mason ◽  
Ron G. Robinson ◽  
Chrysanthe M. Spais ◽  
Mark A. Ator ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Biosynthetic Pathway ◽  
Cellular Assay ◽  
Fatty Acid Biosynthetic Pathway

scholarly journals Regulation of the Omega-3 Fatty Acid Biosynthetic Pathway in Atlantic Salmon Hepatocytes

PLoS ONE ◽  
2016 ◽  
Vol 11 (12) ◽  
pp. e0168230 ◽  
Author(s):  
Marte Avranden Kjær ◽  
Bente Ruyter ◽  
Gerd Marit Berge ◽  
Yajing Sun ◽  
Tone-Kari Knutsdatter Østbye
Keyword(s):  
Fatty Acid ◽  
Atlantic Salmon ◽  
Biosynthetic Pathway ◽  
Omega 3 ◽  
Omega 3 Fatty Acid ◽  
Fatty Acid Biosynthetic Pathway

scholarly journals Cloning and expression of the acyl thioesterase 2 (ALT2) gene in Escherichia coli for producing short- and medium-chain 2-methylketones

2015 ◽  
Vol 18 (2) ◽  
pp. 87-98
Author(s):  
Vy Le Uyen Khuat ◽  
Thao Phuong Do ◽  
Thuong Thi Hong Nguyen
Keyword(s):  
Escherichia Coli ◽  
Food Industry ◽  
Biosynthetic Pathway ◽  
Biodiesel Production ◽  
Cloning And Expression ◽  
Melting Points ◽  
E Coli ◽  
Flame Ionization ◽  
Fatty Acid Biosynthetic Pathway ◽  
Natural Insecticides

2-Methylketones are organic compounds that are more widely used in the food industry and cosmetics. Some of methylketones found in plants act as pheromones and natural insecticides. Recently, methylketones have been recognized as strong candidates for the production of renewable energy as they possess not only favourable cetane numbers but also lower hydrophilicity and melting points than fatty acids which have been used in biodiesel production. In addition, short chain methylketones have much lower melting points than long chain methylketones. In this study, we cloned and expressed in Escherichia coli C41(DE3) cells a gene for acyl thioesterase 2 (ALT2) isolated from Arabidopsis thaliana. The ALT2 enzyme could hydrolyze 3-ketoacyl-ACP (also called β-ketoacyl-ACP) intermediates in the fatty acid biosynthetic pathway into the corresponding 3-ketoacid. The resulting 3- ketoacids are unstable compounds that will be decarboxylated to produce n-1 methylketones. Methylketones released in the growth medium of E. coli expressing ALT2 were extracted by hexane and analyzed by gas chromatography with the flame ionization detector (GC-FID). The results showed that the E. coli C41(DE3) cells expressing ALT2 recombinantly produced 2-nonanone (9C), 2- undecanone (11C) and 2-tridecanone (13C) in the spent medium when were induced with 0.25 mM IPTG at 37 oC for 3.5 hours.

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