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  • Production of P-aminobenzoic acid by metabolically engineered escherichia coli.

Production of P-aminobenzoic acid by metabolically engineered escherichia coli.

Bioscience, biotechnology, and biochemistry (2014-07-19)
Daisuke Koma, Hayato Yamanaka, Kunihiko Moriyoshi, Kiyofumi Sakai, Takaya Masuda, Yoshihiro Sato, Kozo Toida, Takashi Ohmoto
ABSTRACT

The production of chemical compounds from renewable resources is an important issue in building a sustainable society. In this study, Escherichia coli was metabolically engineered by introducing T7lac promoter-controlled aroF(fbr), pabA, pabB, and pabC genes into the chromosome to overproduce para-aminobenzoic acid (PABA) from glucose. Elevating the copy number of chromosomal PT7lac-pabA-pabB distinctly increased the PABA titer, indicating that elevation of 4-amino-4-deoxychorismic acid synthesis is a significant factor in PABA production. The introduction of a counterpart derived from Corynebacterium efficiens, pabAB (ce), encoding a fused PabA and PabB protein, resulted in a considerable increase in the PABA titer. The introduction of more than two copies of PT7lac-pabAB (ce-mod), a codon-optimized pabAB (ce), into the chromosome of a strain that simultaneously overexpressed aroF(fbr) and pabC resulted in 5.1 mM PABA from 55.6 mM glucose (yield 9.2%). The generated strain produced 35 mM (4.8 g L(-1)) PABA from 167 mM glucose (yield 21.0%) in fed-batch culture.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
4-Aminobenzoic acid, ReagentPlus®, 99%
Sigma-Aldrich
4-Aminobenzoic acid, purified by sublimation, ≥99%
Sigma-Aldrich
4-Aminobenzoic acid, ReagentPlus®, ≥99%
Supelco
4-Aminobenzoic acid, analytical standard
4-Aminobenzoic acid, European Pharmacopoeia (EP) Reference Standard