Cloning and expression in Escherichia coli of sucrose phosphorylase gene from Leuconostoc mesenteroides No. 165.
The sucrose phosphorylase gene of an isolate, Leuconostoc mesenteroides No. 165, was amplified by PCR, cloned on pUC118, and expressed in E. coli. The nucleotide sequence of the gene showed 96.3% similarity to that of L. mesenteroides ATCC12291 and 67% to that of Streptococcus mutans, but low similarity to the Agrobacterium vitis gene. The cloned gene, which fusing with lacZalpha, was expressed inducibly with IPTG in E. coli to produce an active enzyme in large quantities that accounted for about 50% of the total cell protein.
Kawasaki
H
H
Department of Applied Biochemistry, Faculty of Agriculture, Osaka Prefecture University, Japan.
Nakamura
N
N
Ohmori
M
M
Sakai
T
T
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D015870:Gene Expression; D005964:Glucosyltransferases / Q000096:biosynthesis* / Q000235:genetics*; D007934:Leuconostoc / Q000201:enzymology* / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid
D001426:Bacterial Proteins; D005964:Glucosyltransferases; C025901:sucrose phosphorylase