Sequential precipitation with reversibly soluble insoluble polymers as a bioseparation strategy: Purification of beta-glucosidase from Trichoderma longibrachiatum

被引:18
作者
Agarwal, R [1 ]
Gupta, MN [1 ]
机构
[1] INDIAN INST TECHNOL,DEPT CHEM,NEW DELHI 110016,INDIA
关键词
D O I
10.1006/prep.1996.0042
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Precipitation of proteins/enzymes with reversibly soluble-insoluble polymers is a simple approach which can be easily scaled up. it is shown here that; considerable purification may be achieved if more than one polymer (chosen judiciously) is used in sequence for purification. The principle is illustrated with purification of beta-glucosidase from a commercial preparation of Trichoderma longibrachiatum. Precipitation with chitosan removed cellulase activity from the preparation with 4-fold purification and 99% recovery of beta-glucosidase activity in the supernatant. The treatment of the supernatant with Eudragit S-100 precipitated 98% of the enzyme activity. The two different elution procedures gave 82% recovery (with 14-fold purification) and 86% recovery (with 12-fold purification) of the enzyme activity from the precipitate. A low molecular weight protein still present in these preparations could be removed by gel filtration. This finally yielded an electrophoretically homogeneous enzyme with 74% recovery and 29-fold purification. (C) 1996 Academic Press, Inc.
引用
收藏
页码:294 / 298
页数:5
相关论文
共 16 条
[1]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[2]   MEASUREMENT OF CELLULASE ACTIVITIES [J].
GHOSE, TK .
PURE AND APPLIED CHEMISTRY, 1987, 59 (02) :257-268
[3]  
Guoquiang D, 1993, BIOSEPARATION, V3, P333
[4]  
Gupta M., 1994, CHEM IND, V17, P673
[5]   PURIFICATION OF XYLANASE FROM TRICHODERMA-VIRIDE BY PRECIPITATION WITH AN ANIONIC POLYMER EUDRAGIT S-100 [J].
GUPTA, MN ;
DONG, GQ ;
KAUL, R ;
MATTIASSON, B .
BIOTECHNOLOGY TECHNIQUES, 1994, 8 (02) :117-122
[6]  
GUPTA MN, 1994, HIGHLY SELECTIVE SEP, P7
[7]   PRODUCTION OF CELLOBIOSE BY ENZYMATIC-HYDROLYSIS - REMOVAL OF BETA-GLUCOSIDASE FROM CELLULASE BY AFFINITY PRECIPITATION USING CHITOSAN [J].
HOMMA, T ;
FUJII, M ;
MORI, J ;
KAWAKAMI, T ;
KURODA, K ;
TANIGUCHI, M .
BIOTECHNOLOGY AND BIOENGINEERING, 1993, 41 (04) :405-410
[8]   MOLECULAR-CLONING AND EXPRESSION OF A BETA-GLUCOSIDASE GENE FROM RUMINOCOCCUS-ALBUS IN ESCHERICHIA-COLI [J].
HONDA, H ;
SAITO, T ;
IIJIMA, S ;
KOBAYASHI, T .
ENZYME AND MICROBIAL TECHNOLOGY, 1988, 10 (09) :559-562
[9]   PARTIAL-PURIFICATION, CHARACTERIZATION AND REGULATION OF CELLULOLYTIC ENZYMES FROM TRICHODERMA-LONGIBRACHIATUM [J].
KALRA, MK ;
SIDHU, MS ;
SANDHU, DK .
JOURNAL OF APPLIED BACTERIOLOGY, 1986, 61 (01) :73-80
[10]  
KUMAR A, 1994, BIOTECHNOL TECH, V8, P651