CLONING, SEQUENCING, AND EXPRESSION OF THE N-ACYL-D-MANNOSAMINE DEHYDROGENASE GENE FROM FLAVOBACTERIUM SP STRAIN-141-8 IN ESCHERICHIA-COLI

被引:14
作者
YAMAMOTOOTAKE, H [1 ]
KOYAMA, Y [1 ]
HORIUCHI, T [1 ]
NAKANO, E [1 ]
机构
[1] NODA INST SCI RES,CHIBA 278,JAPAN
关键词
D O I
10.1128/AEM.57.5.1418-1422.1991
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
The gene coding for N-acyl-D-mannosamine dehydrogenase (NAM-DH) from Flavobacterium sp. strain 141-8 was cloned and expressed under the control of a lac promoter in Escherichia coli JM109. The DNA sequence of the gene was determined, and an open reading frame encoding a polypeptide composed of 272 amino acid residues (M(r), 27,473) was identified. The E. coli transformants which showed over 200-fold higher NAM-DH activity than did the Flavobacterium strain produced the enzyme as a protein fused with beta-galactosidase. Despite being a fusion, NAM-DH produced by E. coli transformants appeared unchanged in pH optimum, K(m), and substrate specificity from Flavobacterium sp. strain 141-8. This newly recombinant enzyme may be applicable to the quantitative determination of sialic acid in serum.
引用
收藏
页码:1418 / 1422
页数:5
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