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嗜碱气单胞菌212号木聚糖酶基因在大肠杆菌中的分子克隆与表达

Molecular cloning and expression of a xylanase gene of alkalophilic Aeromonas sp. no. 212 in Escherichia coli.

作者信息

Kudo T, Ohkoshi A, Horikoshi K

出版信息

J Gen Microbiol. 1985 Oct;131(10):2825-30. doi: 10.1099/00221287-131-10-2825.

Abstract

A gene coding for a xylanase activity of alkalophilic Aeromonas sp. no. 212 (ATCC 31085) was cloned in Escherichia coli HB101 with pBR322. Plasmid pAX1 was isolated from transformants producing xylanase, and the xylanase gene was located in a 6.0 kb Hind III fragment. The pAX1-encoded xylanase activity in E. coli HB101 was about 80 times higher than that of xylanase L in alkalophilic Aeromonas sp. no. 212. About 40% of the enzyme activity was observed in the periplasmic space of E. coli HB101. The pAX1-encoded xylanase had the same enzymic properties as those of xylanase L produced by alkalophilic Aeromonas sp. no. 212, but its molecular weight was lower (135 000 vs 145 000, as estimated by SDS polyacrylamide gel electrophoresis).

摘要

编码嗜碱气单胞菌212号菌株(ATCC 31085)木聚糖酶活性的基因,通过pBR322在大肠杆菌HB101中进行克隆。从产生木聚糖酶的转化体中分离出质粒pAX1,木聚糖酶基因位于一个6.0 kb的Hind III片段中。大肠杆菌HB101中pAX1编码的木聚糖酶活性比嗜碱气单胞菌212号菌株中的木聚糖酶L高约80倍。在大肠杆菌HB101的周质空间中观察到约40%的酶活性。pAX1编码的木聚糖酶具有与嗜碱气单胞菌212号菌株产生的木聚糖酶L相同的酶学性质,但其分子量较低(通过SDS聚丙烯酰胺凝胶电泳估计,分别为135 000和145 000)。

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