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Biochem Biophys Res Commun. 2018 May 5;499(2):279-284. doi: 10.1016/j.bbrc.2018.03.146. Epub 2018 Mar 23.

Characterization of a non-phosphotransferase system for cis,cis-muconic acid production in Corynebacterium glutamicum.

Author information

1
Green Chemistry and Materials Group, Korea Institute of Industrial Technology, Cheonan si, Chungcheongnam-do, 31056, Republic of Korea.
2
Green Chemistry and Materials Group, Korea Institute of Industrial Technology, Cheonan si, Chungcheongnam-do, 31056, Republic of Korea; Green Process and System Engineering Major, Korea University of Science and Technology (UST), Daejeon, 34141, Republic of Korea.
3
STR Biotech Co., Ltd., Bioplaza 4-3, 56, Soyanggang-ro, Chuncheon-si, Gangwon-do, 200-957, Republic of Korea.
4
Department of Molecular Bio-science, Kangwon National University, Chuncheon-si, Gangwon-do, 200-701, Republic of Korea.
5
Department of Biological Engineering, Inha University, Incheon, 402-751, Republic of Korea.
6
Green Chemistry and Materials Group, Korea Institute of Industrial Technology, Cheonan si, Chungcheongnam-do, 31056, Republic of Korea; Green Process and System Engineering Major, Korea University of Science and Technology (UST), Daejeon, 34141, Republic of Korea. Electronic address: sykim@kitech.re.kr.

Abstract

Cis,cis-muconic acid (CCM) is a biochemical material that can be used for the production of various plastics and polymers and is particularly gaining attention as an adipic acid precursor for the synthesis of nylon-6,6. In the current study, the production of CCM was first attempted by introducing a newly developed protocatechuate (PCA) decarboxylase from Corynebacterium glutamicum 13032 to inha103, which completed the biosynthetic pathway therein. To improve CCM productivity, a phosphoenol pyruvate (PEP)-dependent phosphotransferase system (PTS) that consumed the existing glucose was developed, in the form of a strain with a non-PTS that did not consume PEP. To improve glucose uptake, we developed P25 strain, in which iolR (a transcriptional regulator gene) was additionally deleted. Strain P28, a P25 derivative expressing PCA decarboxylase, produced 4.01 g/L of CCM, which was 14% more than that produced by the parental strain. Moreover, strains P29 and P30, with an active pentose phosphate pathway and overexpressing important genes (qsuB) in the metabolic pathway, produced 4.36 and 4.5 g/L of CCM, respectively. Particularly, the yield per glucose in strain P30 was similar to that of the fed-batch culture of Escherichia coli, which has the highest reported yield of 22% (mol/mol). These results are underpinned by the characteristics of the non-PTS with increased PEP availability and a strain with deletion of the iolR gene, which greatly increased glucose uptake.

KEYWORDS:

Carbon flux; Corynebacterium glutamicum; Phosphotransferase system; cis,cis-muconic acid

PMID:
29571737
DOI:
10.1016/j.bbrc.2018.03.146
[Indexed for MEDLINE]
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