通过氨基酸替换提高木聚糖结合结构域的结合活性。

Improvement of binding activity of xylan-binding domain by amino acid substitution.

作者信息

Sakata Tomoko, Takakura Jun, Miyakubo Hiroyuki, Osada Yuko, Wada Rieko, Takahashi Hidenori, Yatsunami Rie, Fukui Toshiaki, Nakamura Satoshi

机构信息

Department of Bioengineering, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama 226-8501, Japan.

出版信息

Nucleic Acids Symp Ser (Oxf). 2006(50):253-4. doi: 10.1093/nass/nrl126.

DOI:10.1093/nass/nrl126

PMID:17150913

Abstract

Xylanase J (XynJ) of alkaliphilic Bacillus sp. strain 41M-1 is a multi-domain enzyme and consists of a glycoside hydrolase (GH) family 11 catalytic domain and an additional xylan-binding domain (XBD) belonging to carbohydrate-binding module (CBM) family 36. Random mutations were introduced into the XBD gene and the repertoire was cloned for display on the surface of filamentous phage. The mutant XBD with amino acid substitution T316I (Thr317 was replaced by Ile) showed higher xylan-binding activity compared to the wild-type XBD. Furthermore, hydrolyzing activity of XynJ toward insoluble xylan was also improved by introducing the mutation T316I.

摘要

嗜碱芽孢杆菌属菌株41M-1的木聚糖酶J（XynJ）是一种多结构域酶，由糖苷水解酶（GH）家族11催化结构域和属于碳水化合物结合模块（CBM）家族36的另一个木聚糖结合结构域（XBD）组成。将随机突变引入XBD基因，并克隆该文库以展示在丝状噬菌体表面。与野生型XBD相比，氨基酸取代T316I（苏氨酸317被异亮氨酸取代）的突变型XBD表现出更高的木聚糖结合活性。此外，通过引入突变T316I，XynJ对不溶性木聚糖的水解活性也得到了提高。