Enhancement of thymidine production in E. coli by eliminating repressors regulating the carbamoyl phosphate synthetase operon.

Abstract:

PURPOSE OF WORK:Thymidine is an important precursor in antiviral drugs. We have enhanced thymidine production in E. coli by eliminating the repressors in the transcription of the gene coding for carbamoyl phosphate synthetase. The operon for carbamoyl phosphate synthetase (CarAB) in the thymidine biosynthesis regulatory pathway was derepressed by disrupting three known repressors (purR, pepA and argR). Combinatorial disruption of three repressors increased CarA expression levels in accordance with degree of disruption, which had a positive correlation with thymidine production. By simultaneous disruption of three repressors (BLdtugRPA), CarA expression level was increased by 3-fold compared to the parental strain, leading to an increased thymidine yield from 0.25 to 1.1 g thymidine l(-1). From BLdtugRPA, we established BLdtugRPA24 by transforming two plasmids expressing enzymes in the thymidine biosynthetic pathway and obtained 5.2 g thymidine l(-1) by Ph-stat fed-batch fermentation.

journal_name

Biotechnol Lett

journal_title

Biotechnology letters

authors

Koo BS,Hyun HH,Kim SY,Kim CH,Lee HC

doi

10.1007/s10529-010-0413-7

subject

Has Abstract

pub_date

2011-01-01 00:00:00

pages

71-8

issue

1

eissn

0141-5492

issn

1573-6776

journal_volume

33

pub_type

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