Analysis of Corynebacterium glutamicum methionine biosynthetic pathway: isolation and analysis of metB encoding cystathionine gamma-synthase.
The metB gene encoding cystathionine y-synthase, the second enzyme of methionine biosynthetic pathway, was isolated from a pSL109-based Corynebacterium glutamicum gene library via complementation of an Escherichia coli metB mutant. A DNA-sequence analysis of the cloned DNA identified an open-reading frame of 1161 bp which encodes a protein with the molecular weight of 41,655 comprising of 386 amino acids. The putative protein product showed good amino acid-sequence homology to its counterpart in other organisms. Introduction of a plasmid carrying the cloned metB into the C. glutamicum resulted in a 10-fold increase in cystathionine gamma-synthase activities, demonstrating the identity of the cloned gene. The C. glutamicum metB mutant which was generated by the site-specific integration of the cloned DNA into its chromosome did not lose the ability to grow on glucose minimal medium lacking supplemental methionine. The growth rate of the mutant strain was also comparable to that of the parental strain. These data indicate that, in addition to the transsulfuration pathway, other methionine biosynthetic pathways may be present in C. glutamicum.
Graduate School of Biotechnology, Korea University, Seoul.
B J
BJ
Hwang
Y
Y
Kim
H B
HB
Kim
H J
HJ
Hwang
J H
JH
Kim
H S
HS
Lee
D000595:Amino Acid Sequence; D001483:Base Sequence; D019757:Carbon-Oxygen Lyases / Q000235:genetics*; D003001:Cloning, Molecular; D003352:Corynebacterium / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism*; D004269:DNA, Bacterial / Q000235:genetics; D015870:Gene Expression; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D008715:Methionine / Q000096:biosynthesis*; D008969:Molecular Sequence Data; D009154:Mutation; D017386:Sequence Homology, Amino Acid
D004269:DNA, Bacterial; D008715:Methionine; C022696:O-succinylhomoserine (thiol)-lyase; D019757:Carbon-Oxygen Lyases