Molecular cloning and characterization of fengycin synthetase gene fenB from Bacillus subtilis

被引:22
作者
Li, GH
Chen, CL
Tschen, JSM
Tsay, SS
Chang, YS
Liu, ST [1 ]
机构
[1] Chang Gung Univ, Dept Microbiol & Immunol, Mol Genet Lab, Tao Yuan 333, Taiwan
[2] Natl Taiwan Univ, Grad Inst Biol, Taipei 106, Taiwan
[3] Natl Chung Hsing Univ, Grad Inst Bot, Taichung 402, Taiwan
关键词
D O I
10.1128/JB.180.5.1338-1341.1998
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
A fengycin synthetase gene, fenB, has been cloned and sequenced. The protein (FenB) encoded by this gene has a predicted molecular mass of 143.6 kDa. This protein was overexpressed in Escherichia coli and was purified to near homogeneity by affinity chromatography. Experimental results indicated that the recombinant FenB has a substrate specificity toward isoleucine with an optimum temperature of 25 degrees C, an optimum pH of 4.5, a K-m value of 922 mu M, and a turnover number of 236 s(-1). FenB also consists of a thioesterase domain, suggesting that this protein mag be involved in the activation of the last amino acid of fengycin.
引用
收藏
页码:1338 / 1341
页数:4
相关论文
共 25 条
[1]  
CHANG LK, 1994, GENE, V150, P129
[2]   TRANSPOSON MUTAGENESIS AND CLONING OF THE GENES ENCODING THE ENZYMES OF FENGYCIN BIOSYNTHESIS IN BACILLUS-SUBTILIS [J].
CHEN, CL ;
CHANG, LK ;
CHANG, YS ;
LIU, ST ;
TSCHEN, JSM .
MOLECULAR & GENERAL GENETICS, 1995, 248 (02) :121-125
[3]   SEQUENCE AND ANALYSIS OF THE GENETIC-LOCUS RESPONSIBLE FOR SURFACTIN SYNTHESIS IN BACILLUS-SUBTILIS [J].
COSMINA, P ;
RODRIGUEZ, F ;
DEFERRA, F ;
GRANDI, G ;
PEREGO, M ;
VENEMA, G ;
VANSINDEREN, D .
MOLECULAR MICROBIOLOGY, 1993, 8 (05) :821-831
[4]  
DECRECYLAGARD V, 1995, LIFE SCI, V318, P927
[5]  
DEFERRA F, 1997, J BIOL CHEM, V272, P25307
[6]   Substrate specificity of hybrid modules from peptide synthetases [J].
Elsner, A ;
Engert, H ;
Saenger, W ;
Hamoen, L ;
Venema, G ;
Bernhard, F .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1997, 272 (08) :4814-4819
[7]   ANALYSIS OF CORE SEQUENCES IN THE D-PHE ACTIVATING DOMAIN OF THE MULTIFUNCTIONAL PEPTIDE SYNTHETASE TYCA BY SITE-DIRECTED MUTAGENESIS [J].
GOCHT, M ;
MARAHIEL, MA .
JOURNAL OF BACTERIOLOGY, 1994, 176 (09) :2654-2662
[8]   NONRIBOSOMAL BIOSYNTHESIS OF PEPTIDE ANTIBIOTICS [J].
KLEINKAUF, H ;
VONDOHREN, H .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1990, 192 (01) :1-15
[9]   A nonribosomal system of peptide biosynthesis [J].
Kleinkauf, H ;
VonDohren, H .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1996, 236 (02) :335-351
[10]  
KOCH U, 1988, THESIS E KARLS U TUB