Cloning and expression of the sucrose phosphorylase gene from Leuconostoc mesenteroides in Escherichia coli
Cloning and expression of the sucrose phosphorylase gene from Leuconostoc mesenteroides in Escherichia coli
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DOI：
10.1007/s10529-007-9608-y
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发表时间：
2008-04-01
影响因子：
2.7
通讯作者：
Kim, Doman
中科院分区：
文献类型：
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作者：
Lee, Jin-Ha;Moon, Young-Hwan;Kim, Doman
克隆肠膜明串珠菌NRRL B-742蔗糖磷酸化酶基因（742 sp），并在大肠杆菌中表达。转化的742 sp的核苷酸序列包含1，458 bp的ORF，得到计算分子量为55.3kDa的蛋白质。742 SPase的C-末端氨基酸序列与其它Leu的C-末端氨基酸序列显著不同。肠系膜SPases.纯化的742 SPase比活性为1.8 U/mg，以蔗糖为底物时的Km为3 mM;最佳活性为37 ℃和pH 6.7。纯化的742 SPase将蔗糖的葡萄糖基部分转移到胞嘧啶一磷酸（CMP）。
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).