生物技术通报 ›› 2020, Vol. 36 ›› Issue (9): 235-243.doi: 10.13560/j.cnki.biotech.bull.1985.2020-0079

• 研究报告 • 上一篇    下一篇

生长抑素和耐热木聚糖酶的融合表达及性质研究

黄坤龙, 苏小运, 姚斌   

  1. 中国农业科学院饲料研究所 农业部饲料生物技术重点开放实验室,北京 100081
  • 收稿日期:2020-01-02 出版日期:2020-09-26 发布日期:2020-09-30
  • 作者简介:黄坤龙,男,硕士,研究方向:生物化学和分子生物学;E-mail:302845038@139.com
  • 基金资助:
    国家肉鸡产业技术体系(CARS-41),国家自然科学基金面上项目(31672458),中国农业科学院饲料研究所基本科研业务费

Fusion Expression of Somatostatin with a Thermostable Xylanase and Characterization of the Fusion Protein

HUANG Kun-long, SU Xiao-yun, YAO Bin   

  1. Key Laboratory of Feed Biotechnology of Ministry of Agriculture,Feed Research Institute,Chinese Academy of Agricultural Sciences,Beijing 100081
  • Received:2020-01-02 Published:2020-09-26 Online:2020-09-30

摘要: 旨在获得具有抗原性和免疫原性的融合蛋白CbXyn10C-SS,并对其进行高效表达和酶学性质分析。生长抑素(Somatostatin,SS)从多途径阻碍饲料转化效率和动物生长,使用SS免疫动物可避免其抑制作用。然而SS免疫的研究多以DNA疫苗为载体,少见重组功能性SS蛋白的高效制备方法。木聚糖酶提高饲料转化率并在饲料中广泛应用,且微生物法发酵制备重组木聚糖酶技术已经非常成熟。将SS基因连接在细菌Caldicellulosiruptor bescii来源的耐热木聚糖酶基因CbXyn10C的3'端进行融合表达。融合蛋白CbXyn10C-SS具有抗原性和免疫原性,且该蛋白的理化性质和催化常数与CbXyn10C非常相似,具高度的热稳定性和较强的催化性能,适合饲料制粒。成功高效表达了能同时作为饲用木聚糖酶和生长抑素抗原的融合蛋白CbXyn10C-SS。

关键词: 生长抑素, 木聚糖酶, 融合表达, 抗原表位

Abstract: The aim of this study is to obtain the fusion protein CbXyn10C-SS with antigenicity and immunogenicity,to efficiently express it,and to characterize the enzymatic properties. Somatostatin(SS)impedes feed transformation efficiency and animal growth in multiple ways. Therefore,using SS to immunize animals can avoid the inhibitory effect. However,the research of SS immunization mostly uses DNA vaccine as the carrier and highly efficient recombinant production of SS protein is seldom reported. Xylanase is widely used in feed,which can improve feed conversion rate. Till now,recombinant xylanase has been successfully expressed in microbial cells. The SS gene was fused to the 3' terminus of the thermostable xylanase gene CbXyn10C derived from the Caldicellulosiruptor bescii. The fusion protein CbXyn10C-SS presented antigenicity and immunogenicity. Being highly thermostable and efficient in catalysis,CbXyn10C-SS also demonstrated very similar biochemical characteristics and catalytic parameters to those of CbXyn10C,which is therefore suitable for the feed pelleting process. The successfully and efficiently expressed fusion protein CbXyn10C-SS has potential application value as a feed xylanase and somatostatin antigen.

Key words: somatostatin, xylanase, fusion expression, antigenic epitope