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

来源：

APPLIED AND ENVIRONMENTAL MICROBIOLOGY | 1991年 / 57卷 / 05期

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

页码：1418 / 1422

页数：5

共 17 条

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