Please wait a minute...
 
 
食品与发酵工业  2017, Vol. 43 Issue (6): 8-    
  本期目录 | 过刊浏览 | 高级检索 |
酿酒酵母孢子表面展示系统的构建及应用
乔颍鑫,李子杰,中西秀树,高晓冬
江南大学生物工程学院,糖化学与生物技术教育部重点实验室,江苏无锡,214122
Construction and application of a novel surface display system on yeast spore
QIAO Ying-xin,LI Zi-jie,NAKANISHI Hideki,GAO Xiao-dong
下载:  PDF (2167KB) 
输出:  BibTeX | EndNote (RIS)      
摘要 旨在建立1种酿酒酵母孢子表面展示系统,并将其应用于稀有糖合成.表面展示系统是运用微生物自身功能把外源蛋白或多肽展示在细胞表面,文中用磷酸甘油氧化酶(GPO)来验证酿酒酵母孢子表面展示系统的可行性.将编码肺炎链球菌来源的GPO的基因glpo与信号肽序列(ss)融合后连接到载体pRS42 4-TE Fpr上,将重组质粒pR S42 4-TEFpr-ss-glpo转化至酿酒酵母野生型及缺陷型菌株中并进行产孢,通过荧光观察、western blot分析以及酶活测定,表明酿酒酵母缺陷型孢子可以实现蛋白的表面展示.通过对GPO孢子表征,表明GPO孢子在pH为7.0、温度为30℃时活性最高,该孢子作为催化剂可以重复使用,当使用第3次时,活性可达最大活性的40%.最后将GPO孢子应用于“一锅四酶法”进行稀有糖L-果糖的合成,转化率为12%.
服务
把本文推荐给朋友
加入引用管理器
E-mail Alert
RSS
作者相关文章
Abstract: In this paper,a novel surface display system on yeast spore and its application for rare sugar synthesis were studied.Surface display system was used by microorganisms to display the exogenous proteins or peptides on their cell surface.Glycerophosphate oxidase (GPO) was used to verify the feasibility of the novel surface display system.The gene glpo GPO from Streptococcus pneumoniae was fused with signal sequenceand inserted into pRS424-TE-Fpr vector.The recombinant plasmid pRS424-TEFpr-ss-glpo was transformed into wild type and auxotroph of S.cerevisiae to sporulate.The results from fluorescence analysis,western blot detection and activity assays showed that S.cerevisiae spore surface display system was successfully constructed.The GPO spores were characterized and their optimum pH and temperature were 7.0 and 30℃,respectively.Moreover,the spores could be reused with activity of about 40% of the maximum activity at the third time.Finally,GPO spores were employed in the one-pot four-enzyme system for synthesis of rare sugar L-fructose with yield of about 12%.
               出版日期:  2017-06-25      发布日期:  2017-12-01      期的出版日期:  2017-06-25
基金资助: 国家自然科学基金(21302069)
中央高校基本科研业务费专项基金资助(JUSRP1003)
教育部科学技术研究重大项目(313027)
引用本文:    

链接本文:  
http://sf1970.cnif.cn/CN/  或          http://sf1970.cnif.cn/CN/Y2017/V43/I6/8
No related articles found!
No Suggested Reading articles found!
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
版权所有 © 《食品与发酵工业》编辑部
地址:北京朝阳区酒仙桥中路24号院6号楼111室
本系统由北京玛格泰克科技发展有限公司设计开发  技术支持:support@magtech.com.cn