嗜热菌“解糖热纤梭菌”中木聚糖降解酶编码基因的克隆、序列分析及表达
Cloning, sequence analysis, and expression of genes encoding xylan-degrading enzymes from the thermophile "Caldocellum saccharolyticum".
作者信息
Lüthi E, Love D R, McAnulty J, Wallace C, Caughey P A, Saul D, Bergquist P L
机构信息
Department of Cellular and Molecular Biology, University of Auckland, New Zealand.
出版信息
Appl Environ Microbiol. 1990 Apr;56(4):1017-24. doi: 10.1128/aem.56.4.1017-1024.1990.
Abstract
A lambda recombinant bacteriophage coding for xylanase and beta-xylosidase activity has been isolated from a genomic library of the extremely thermophilic anaerobe "Caldocellum saccharolyticum." Partial Sau3AI fragments of the lambda recombinant DNA were ligated into pBR322. A recombinant plasmid with an insertion of ca. 7 kilobases of thermophilic DNA expressing both enzymatic activities was isolated. The location of the genes has been established by analyzing deletion derivatives, and the DNA sequence of 6.067 kilobases of the insert has been determined. Five open reading frames (ORFs) were found, one of which (ORF1; Mr 40,455) appears to code for a xylanase (XynA) which also acts on o-nitrophenyl-beta-D-xylopyranoside. Another, ORF5 (Mr 56,365), codes for a beta-xylosidase (XynB). The xynA gene product shows significant homology with the xylanases from the alkalophilic Bacillus sp. strain C125 and Clostridium thermocellum.
摘要
从嗜热厌氧菌“嗜糖栖热放线菌”的基因组文库中分离出一种编码木聚糖酶和β-木糖苷酶活性的λ重组噬菌体。λ重组DNA的部分Sau3AI片段被连接到pBR322中。分离出一个插入了约7千碱基嗜热DNA且表达两种酶活性的重组质粒。通过分析缺失衍生物确定了基因的位置，并测定了插入片段6.067千碱基的DNA序列。发现了五个开放阅读框（ORF），其中一个（ORF1；分子量40,455）似乎编码一种也作用于邻硝基苯基-β-D-木吡喃糖苷的木聚糖酶（XynA）。另一个ORF5（分子量56,365）编码一种β-木糖苷酶（XynB）。xynA基因产物与嗜碱芽孢杆菌属菌株C125和嗜热栖热梭菌的木聚糖酶具有显著同源性。
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