Cloning and expression of the sucrose phosphorylase gene from Leuconostoc mesenteroides in Escherichia coli.
The gene encoding sucrose phosphorylase (742sp) in Leuconostoc mesenteroides NRRL B-742 was cloned and expressed in Escherichia coli. The nucleotide sequence of the transformed 742sp comprised an ORF of 1,458 bp giving a protein with calculated molecular mass of 55.3 kDa. 742SPase contains a C-terminal amino acid sequence that is significantly different from those of other Leu. mesenteroides SPases. The purified 742SPase had a specific activity of 1.8 U/mg with a K (m) of 3 mM with sucrose as a substrate; optimum activity was at 37 degrees C and pH 6.7. The purified 742SPase transferred the glucosyl moiety of sucrose to cytosine monophosphate (CMP).
Dental Science Research Institute, School of Dentistry, 2nd Stage of Brain Korea 21 for School of Dentistry, Chonnam National University, Gwang-ju 500-757, South Korea.
Jin-Ha
JH
Lee
Young-Hwan
YH
Moon
Nahyun
N
Kim
Young-Min
YM
Kim
Hee-Kyoung
HK
Kang
Ji-Yeon
JY
Jung
Emad
E
Abada
Seong-Soo
SS
Kang
Doman
D
Kim
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002855:Chromatography, Thin Layer; D003001:Cloning, Molecular / Q000379:methods; D003568:Cytidine Monophosphate / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D005964:Glucosyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D007934:Leuconostoc / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D015394:Molecular Structure; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D013395:Sucrose / Q000378:metabolism
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D013395:Sucrose; D005964:Glucosyltransferases; C025901:sucrose phosphorylase; D003568:Cytidine Monophosphate