Purification, characterization and identification of cysteine desulfhydrase of Corynebacterium glutamicum, and its relationship to cysteine production.
We highly purified the enzyme having L-cysteine desulfhydrase activity from Corynebacterium glutamicum. According to its partial amino acid sequence, the enzyme was identified as the aecD gene product, a C-S lyase with alpha, beta-elimination activity [I. Rossol and A. Pühler (1992) J. Bacteriol. 174, 2968-2977]. To produce L-cysteine in C. glutamicum, the Escherichia coli-altered cysE gene encoding Met256Ile mutant serine acetyltransferase, which is desensitized to feedback inhibition by L-cysteine, was introduced into C. glutamicum. When the altered cysE gene was expressed in the aecD-disrupted strain, the transformants produced approximately 290 mg of L-cysteine plus L-cystine per liter from glucose. The produced amount of these amino acids was about two-fold higher than that of the wild-type strain.
Wada
Masaru
M
Department of Bioscience, Fukui Prefectural University, 4-1-1 Kenjyojima, Matsuoka-cho, Fukui 910-1195, Japan. masaru@fpu.ac.jp
Awano
Naoki
N
Haisa
Keiko
K
Takagi
Hiroshi
H
Nakamori
Shigeru
S
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D003352:Corynebacterium / Q000201:enzymology* / Q000235:genetics; D003542:Cystathionine gamma-Lyase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D003545:Cysteine / Q000032:analysis / Q000096:biosynthesis* / Q000378:metabolism; D003553:Cystine / Q000032:analysis / Q000096:biosynthesis / Q000378:metabolism; D008954:Models, Biological; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D020539:Sequence Analysis, Protein; D013379:Substrate Specificity
D003553:Cystine; D003542:Cystathionine gamma-Lyase; D003545:Cysteine