STRATEGY AND IMPLEMENTATION OF A SYSTEM FOR PROTEIN ENGINEERING

被引:10
作者
GODDETTE, DW [1 ]
CHRISTIANSON, T [1 ]
LADIN, BF [1 ]
LAU, M [1 ]
MIELENZ, JR [1 ]
PAECH, C [1 ]
REYNOLDS, RB [1 ]
YANG, SS [1 ]
WILSON, CR [1 ]
机构
[1] COGNIS INC, 2330 CIRCADIAN WAY, SANTA ROSA, CA 95407 USA
关键词
PROTEIN ENGINEERING; THERMOSTABILITY; SUBTILISIN BL; ALKALINE PROTEASE;
D O I
10.1016/0168-1656(93)90124-6
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
This paper describes an overall view of an industrial protein engineering project from conception to successful completion. The choice of rational design was determined by the availability of an excellent three-dimensional crystal structure and the availability of information in the literature to define a strategy. The design strategy was refined extensively during the course of the project. The development of methods for mutagenesis, expression, verification, purification, and characterization of mutant enzymes is dictated in part by the enzyme property one chooses to modify and must be rapid yet accurate. Such an approach would be applicable to improve the stability of any other protein or enzyme. Using this approach, we successfully increased the stability of subtilisin BL over 10-fold at 50-degrees-C with an overall success rate greater than 60%.
引用
收藏
页码:41 / 54
页数:14
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