Synthesis of poly (γ-glutamic acid) and heterologous expression of pgsBCA genes

被引：34

作者：

Cao, Mingfeng ^{[1
]}

Song, Cunjiang ^{[1
]}

Jin, Yinghong ^{[1
,2
]}

Liu, Li ^{[1
,2
]}

Liu, Jing ^{[1
,2
]}

Xie, Hui ^{[1
]}

Guo, Wenbin ^{[1
]}

Wang, Shufang ^{[2
]}

机构：

[1] Nankai Univ, Key Lab Mol Microbiol & Technol, Minist Educ, Coll Life Sci, Tianjin 300071, Peoples R China

[2] Nankai Univ, Key Lab Bioact Mat, Minist Educ, Tianjin 300071, Peoples R China

来源：

JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC | 2010年 / 67卷 / 1-2期

关键词：

Poly (gamma-glutamic acid); Bacillus licheniformis; pgsBCA genes; Shuttle vector pXMJ19; POLYGLUTAMIC ACID; POLY(GAMMA-GLUTAMIC ACID); ESCHERICHIA-COLI; BACILLUS; SYNTHETASE; PRODUCER; CLONING;

D O I：

10.1016/j.molcatb.2010.07.014

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The genes required for synthesis of poly (gamma-glutamic acid) (gamma-PGA) were cloned from Bacillus licheniformis NK-03, a strain isolated from fermented food, natto. There were three open reading frames pgsB, pgsC, pgsA in the cloned fragment, all of which were greatly similar with those from typical Bacillus subtilis strains. The alignment of deduced amino acid sequences showed that PgsC was the most conservative part in PgsBCA. Recombinant plasmid pXMJ19-PGS was constructed by a shuttle vector pXMJ19, and it was successfully transformed and expressed in the recombinant strains of Escherichia coli JM109 and Corynebacterium glutamicum ATCC13032, respectively. Expression of pgsBCA in C. glutamicum indicated that it could synthesize gamma-PGA with a yield of 0.69 g/L and 97% proportion of L-glutamate monomer in the absence of glutamic acid. The results suggest that gamma-PGA biosynthesis directly from glucose by genetic engineering is feasible and significant. (C) 2010 Elsevier B.V. All rights reserved.

引用

页码：111 / 116

页数：6

共 32 条

[1]

ADAMSON JG, 1992, ANAL BIOCHEM, V202, P210

[2]

[Anonymous], 1989, Molecular Cloning: A Laboratory

[3] A poly-γ-glutamate synthetic system of Bacillus subtilis IFO 3336:: Gene cloning and biochemical analysis of poly-γ-glutamate produced by Escherichia coli clone cells [J].

Ashiuchi, M ;

Soda, K ;

Misono, H .

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1999, 263 (01) :6-12

[4] Enzymatic synthesis of high-molecular-mass poly-γ-glutamate and regulation of its stereochemistry [J].

Ashiuchi, M ;

Shimanouchi, K ;

Nakamura, H ;

Kamei, T ;

Soda, K ;

Park, C ;

Sung, MH ;

Misono, H .

APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2004, 70 (07) :4249-4255

[5] Poly-γ-glutamate synthetase of Bacillus subtilis [J].

Ashiuchi, M ;

Kamei, T ;

Misono, H .

JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC, 2003, 23 (2-6) :101-106

[6] Isolation of Bacillus subtilis (chungkookjang), a poly-γ-glutamate producer with high genetic competence [J].

Ashiuchi, M ;

Kamei, T ;

Baek, DH ;

Shin, SY ;

Sung, MH ;

Soda, K ;

Yagi, T ;

Misono, H .

APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2001, 57 (5-6) :764-769

[7] Biochemistry and molecular genetics of poly-γ-glutamate synthesis [J].

Ashiuchi, M ;

Misono, H .

APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2002, 59 (01) :9-14

[8] Physiological and biochemical characteristics of poly γ-glutamate synthetase complex of Bacillus subtilis [J].

Ashiuchi, M ;

Nawa, C ;

Kamei, T ;

Song, JJ ;

Hong, SP ;

Sung, MH ;

Soda, K ;

Yagi, T ;

Misono, H .

EUROPEAN JOURNAL OF BIOCHEMISTRY, 2001, 268 (20) :5321-5328

[9] Genetically engineered poly-γ-glutamate producer from Bacillus subtilis ISW1214 [J].

Ashiuchi, Makoto ;

Shimanouchi, Kazuya ;

Horiuchi, Terumi ;

Kame, Tohru ;

Misono, Haruo .

BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 2006, 70 (07) :1794-1797

[10] A statistical approach to optimization of fermentative production of poly(γ-glutamic acid) from Bacillus licheniformis NCIM 2324 [J].

Bajaj, I. B. ;

Lele, S. S. ;

Singhal, R. S. .

BIORESOURCE TECHNOLOGY, 2009, 100 (02) :826-832

← 1 2 3 4 →