Thermostable glycerol kinase from Thermus flavus: cloning, sequencing, and expression of the enzyme gene.
The thermostable glycerol kinase (EC 2.7.1.30) gene from Thermus flavus was cloned and expressed in Escherichia coli DH5 alpha. An open reading frame of 1488 bp for the glycerol kinase gene (glpK) starting with an ATG methionine codon was found, which encodes a protein of 496 amino acid residues whose calculated molecular weight is 54,835. The amino acid sequence of T. flavus glycerol kinase is 80.6% and 64.1% identical with those of Bacillus subtilis and E. coli. Transformants of E. coli DH5 alpha harboring plasmid pGYK12 with a 1505 bp chromosomal DNA fragment containing the T. flavus glycerol kinase gene showed about 23.8-fold higher glycerol kinase activity than T. flavus.
Huang
H S
HS
School of Pharmaceutical Sciences, Nagasaki University, Japan.
Kabashima
T
T
Ito
K
K
Yin
C H
CH
Nishiya
Y
Y
Kawamura
Y
Y
Yoshimoto
T
T
D000595:Amino Acid Sequence; D019843:Archaeal Proteins / Q000737:chemistry; D001483:Base Sequence; D002729:Chloromercuribenzoates / Q000494:pharmacology; D003001:Cloning, Molecular; D016384:Consensus Sequence / Q000235:genetics; D004795:Enzyme Stability; D015870:Gene Expression / Q000235:genetics; D005991:Glycerol Kinase / Q000737:chemistry*; D008969:Molecular Sequence Data; D011994:Recombinant Proteins; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013824:Thermus / Q000201:enzymology*; D020245:p-Chloromercuribenzoic Acid
D019843:Archaeal Proteins; D002729:Chloromercuribenzoates; D011994:Recombinant Proteins; D020245:p-Chloromercuribenzoic Acid; D005991:Glycerol Kinase