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).
Lee
Jin-Ha
JH
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.
Moon
Young-Hwan
YH
Kim
Nahyun
N
Kim
Young-Min
YM
Kang
Hee-Kyoung
HK
Jung
Ji-Yeon
JY
Abada
Emad
E
Kang
Seong-Soo
SS
Kim
Doman
D
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