Biotin protein ligase from Corynebacterium glutamicum: role for growth and l-lysine production

被引:0
作者
P. Peters-Wendisch
K. C. Stansen
S. Götker
V. F. Wendisch
机构
[1] Bielefeld University,Genetics of Prokaryotes, Faculty of Biology and CeBiTec
来源
Applied Microbiology and Biotechnology | 2012年 / 93卷
关键词
Biotin protein ligase; BirA; Amino acid production; -lysine;
D O I
暂无
中图分类号
学科分类号
摘要
Corynebacterium glutamicum is a biotin auxotrophic Gram-positive bacterium that is used for large-scale production of amino acids, especially of l-glutamate and l-lysine. It is known that biotin limitation triggers l-glutamate production and that l-lysine production can be increased by enhancing the activity of pyruvate carboxylase, one of two biotin-dependent proteins of C. glutamicum. The gene cg0814 (accession number YP_225000) has been annotated to code for putative biotin protein ligase BirA, but the protein has not yet been characterized. A discontinuous enzyme assay of biotin protein ligase activity was established using a 105aa peptide corresponding to the carboxyterminus of the biotin carboxylase/biotin carboxyl carrier protein subunit AccBC of the acetyl CoA carboxylase from C. glutamicum as acceptor substrate. Biotinylation of this biotin acceptor peptide was revealed with crude extracts of a strain overexpressing the birA gene and was shown to be ATP dependent. Thus, birA from C. glutamicum codes for a functional biotin protein ligase (EC 6.3.4.15). The gene birA from C. glutamicum was overexpressed and the transcriptome was compared with the control strain revealing no significant gene expression changes of the bio-genes. However, biotin protein ligase overproduction increased the level of the biotin-containing protein pyruvate carboxylase and entailed a significant growth advantage in glucose minimal medium. Moreover, birA overexpression resulted in a twofold higher l-lysine yield on glucose as compared with the control strain.
引用
收藏
页码:2493 / 2502
页数:9
相关论文
共 334 条
[1]  
Abe S(1967)Taxonomical studies on glutamic acid producing bacteria J Gen Appl Microbiol 13 279-301
[2]  
Takayarna K(2011)Control of J Biotechnol 152 77-86
[3]  
Kinoshita S(2005) and J Mol Biol 353 322-333
[4]  
Auchter M(1981) expression by the SucR regulator in J Mol Biol 146 451-467
[5]  
Laslo T(1981)Crystal structures of biotin protein ligase from J Mol Biol 146 469-492
[6]  
Fleischer C(1999) OT3 and its complexes: structural basis of biotin activation J Biol Chem 274 31767-31769
[7]  
Schiller L(1996)The J Bacteriol 178 4122-4130
[8]  
Arndt A(1986) gene of Gene 44 255-261
[9]  
Gaigalat L(2009) encodes a biotin holoenzyme synthetase J Biotechnol 143 173-182
[10]  
Kalinowski J(1999)Genetic and biochemical characterization of the J Biol Chem 274 1449-1457