Production of p-amino-L-phenylalanine (L-PAPA) from glycerol by metabolic grafting of Escherichia coli

Microbial Cell Factories
Behrouz Mohammadi NargesiJung-Won Youn

Abstract

The non-proteinogenic aromatic amino acid, p-amino-L-phenylalanine (L-PAPA) is a high-value product with a broad field of applications. In nature, L-PAPA occurs as an intermediate of the chloramphenicol biosynthesis pathway in Streptomyces venezuelae. Here we demonstrate that the model organism Escherichia coli can be transformed with metabolic grafting approaches to result in an improved L-PAPA producing strain. Escherichia coli K-12 cells were genetically engineered for the production of L-PAPA from glycerol as main carbon source. To do so, genes for a 4-amino-4-deoxychorismate synthase (pabAB from Corynebacterium glutamicum), and genes encoding a 4-amino-4-deoxychorismate mutase and a 4-amino-4-deoxyprephenate dehydrogenase (papB and papC, both from Streptomyces venezuelae) were cloned and expressed in E. coli W3110 (lab strain LJ110). In shake flask cultures with minimal medium this led to the formation of ca. 43 ± 2 mg l-1 of L-PAPA from 5 g l-1 glycerol. By expression of additional chromosomal copies of the tktA and glpX genes, and of plasmid-borne aroFBL genes in a tyrR deletion strain, an improved L-PAPA producer was obtained which gave a titer of 5.47 ± 0.4 g l-1 L-PAPA from 33.3 g l-1 glycerol (0.16 g L-PAPA/g of glyc...Continue Reading

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Citations

Jan 22, 2019·Frontiers in Bioengineering and Biotechnology·Behrouz Mohammadi NargesiJung-Won Youn
Nov 2, 2019·Nature Communications·Navaneethan PalanisamyBarbara Di Ventura
Dec 30, 2021·Journal of Agricultural and Food Chemistry·Feifei HouWei Huang

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