Improved pinocembrin production in Escherichia coli by engineering fatty acid synthesis. 2016

Weijia Cao, and Weichao Ma, and Bowen Zhang, and Xin Wang, and Kequan Chen, and Yan Li, and Pingkai Ouyang
State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing, 211816, Jiangsu, China.

The development of efficient microbial processes for pinocembrin production has attracted considerable attention. However, pinocembrin biosynthetic efficiency is greatly limited by the low availability of the malonyl-CoA cofactor in Escherichia coli. Fatty acid biosynthesis is the only metabolic process in E. coli that consumes malonyl-CoA; therefore, we overexpressed the fatty acid biosynthetic pathway enzymes β-ketoacyl-ACP synthase III (FabH) and β-ketoacyl-ACP synthase II (FabF) alone and in combination, and investigated the effect on malonyl-CoA. Interestingly, overexpressing FabH, FabF or both enzymes in E. coli BL21 (DE3) decreased fatty acid synthesis and increased cellular malonyl-CoA levels 1.4-, 1.6-, and 1.2-fold, respectively. Furthermore, pinocembrin production was increased 10.6-, 31.8-, and 5.87-fold in recombinant strains overexpressing FabH, FabF and both enzymes, respectively. Overexpression of FabF, therefore, triggered the highest pinocembrin production and malonyl-CoA levels. The addition of cerulenin further increased pinocembrin production in the FabF-overexpressing strain, from 25.8 to 29.9 mg/L. These results demonstrated that overexpressing fatty acid synthases can increase malonyl-CoA availability and improve pinocembrin production in a recombinant E. coli host. This strategy may hold promise for the production of other important natural products in which cellular malonyl-CoA is rate limiting.

UI MeSH Term Description Entries
D008316 Malonyl Coenzyme A A coenzyme A derivative which plays a key role in the fatty acid synthesis in the cytoplasmic and microsomal systems. Malonyl CoA,CoA, Malonyl,Coenzyme A, Malonyl
D004926 Escherichia coli A species of gram-negative, facultatively anaerobic, rod-shaped bacteria (GRAM-NEGATIVE FACULTATIVELY ANAEROBIC RODS) commonly found in the lower part of the intestine of warm-blooded animals. It is usually nonpathogenic, but some strains are known to produce DIARRHEA and pyogenic infections. Pathogenic strains (virotypes) are classified by their specific pathogenic mechanisms such as toxins (ENTEROTOXIGENIC ESCHERICHIA COLI), etc. Alkalescens-Dispar Group,Bacillus coli,Bacterium coli,Bacterium coli commune,Diffusely Adherent Escherichia coli,E coli,EAggEC,Enteroaggregative Escherichia coli,Enterococcus coli,Diffusely Adherent E. coli,Enteroaggregative E. coli,Enteroinvasive E. coli,Enteroinvasive Escherichia coli
D005227 Fatty Acids Organic, monobasic acids derived from hydrocarbons by the equivalent of oxidation of a methyl group to an alcohol, aldehyde, and then acid. Fatty acids are saturated and unsaturated (FATTY ACIDS, UNSATURATED). (Grant & Hackh's Chemical Dictionary, 5th ed) Aliphatic Acid,Esterified Fatty Acid,Fatty Acid,Fatty Acids, Esterified,Fatty Acids, Saturated,Saturated Fatty Acid,Aliphatic Acids,Acid, Aliphatic,Acid, Esterified Fatty,Acid, Saturated Fatty,Esterified Fatty Acids,Fatty Acid, Esterified,Fatty Acid, Saturated,Saturated Fatty Acids
D015099 3-Oxoacyl-(Acyl-Carrier-Protein) Synthase An enzyme of long-chain fatty acid synthesis, that adds a two-carbon unit from malonyl-(acyl carrier protein) to another molecule of fatty acyl-(acyl carrier protein), giving a beta-ketoacyl-(acyl carrier protein) with the release of carbon dioxide. EC 2.3.1.41. 3 Oxoacyl (Acyl Carrier Protein) Synthase,3-Keto-ACP Synthase,3-Oxoacyl (Acyl Carrier Protein) Synthase,3-Oxoacyl Synthetase,Acyl-Malonyl-ACP Condensing Enzyme,beta Keto Acyl Synthetase,beta Keto-Acyl Carrier Protein Synthase I,beta Keto-Acyl Carrier Protein Synthase II,beta Ketoacyl ACP Synthase,beta-Ketoacyl-Coenzyme A (CoA) Synthase,3 Keto ACP Synthase,3 Oxoacyl Synthetase,Acyl Malonyl ACP Condensing Enzyme,Condensing Enzyme, Acyl-Malonyl-ACP,Synthase, 3-Keto-ACP,Synthetase, 3-Oxoacyl,beta Keto Acyl Carrier Protein Synthase I,beta Keto Acyl Carrier Protein Synthase II
D044950 Flavanones A group of FLAVONOIDS characterized with a 4-ketone. 2-Phenyl-Benzopyran-4-Ones,2 Phenyl Benzopyran 4 Ones
D053898 Biosynthetic Pathways Sets of enzymatic reactions occurring in organisms and that form biochemicals by making new covalent bonds. Biosynthetic Pathway,Pathway, Biosynthetic,Pathways, Biosynthetic
D060847 Metabolic Engineering Methods and techniques used to genetically modify cells' biosynthetic product output and develop conditions for growing the cells as BIOREACTORS. Engineering, Metabolic
D019149 Bioreactors Tools or devices for generating products using the synthetic or chemical conversion capacity of a biological system. They can be classical fermentors, cell culture perfusion systems, or enzyme bioreactors. For production of proteins or enzymes, recombinant microorganisms such as bacteria, mammalian cells, or insect or plant cells are usually chosen. Fermentors,Bioreactor,Fermentor
D064429 Fatty Acid Synthases Enzymes that catalyze the synthesis of FATTY ACIDS from acetyl-CoA and malonyl-CoA derivatives. Fatty Acid Synthase,Fatty Acid Synthetases,Acid Synthase, Fatty,Acid Synthases, Fatty,Synthase, Fatty Acid,Synthetases, Fatty Acid

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