QUANTIFICATION AND IDENTIFICATION OF THE MAIN COMPONENTS OF THE TRICHODERMA CELLULASE COMPLEX WITH MONOCLONAL-ANTIBODIES USING AN ENZYME-LINKED-IMMUNOSORBENT-ASSAY (ELISA)

被引:52
作者
KOLBE, J [1 ]
KUBICEK, CP [1 ]
机构
[1] VIENNA TECH UNIV, INST BIOCHEM TECHNOL & MIKROBIOL, MIKROBIELLE BIOCHEM ABT, A-1060 VIENNA, AUSTRIA
关键词
D O I
10.1007/BF00170918
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
An enzyme-linked immunosorbent assay (ELISA) using monoclonal antibodies has been developed to measure the concentration of three main cellulase components from Trichoderma reesei, cellobiohydrolase I (CBH I), cellobiohydrolase II (CBH II) and I (EG I), in both commercial enzyme preparations as well as in samples from laboratory fermentations. The sensitivity of the assay is 1-10 ng protein, depending on the type of cellulase. The coefficient of variability is between 10% and 20%. By a combination of two different domain-specific monoclonals against CBH I or II it is also possible to quantify the concentration of intact and truncated forms of these two enzymes, respectively. The use of the ELISA to quantify the formation of the three cellulase components under different cultivation conditions is described. © 1990 Springer-Verlag.
引用
收藏
页码:26 / 30
页数:5
相关论文
共 27 条
[1]  
[Anonymous], METHOD ENZYMOL
[2]   A METHOD FOR THE DETECTION AND DIFFERENTIATION OF CELLULASE COMPONENTS IN POLYACRYLAMIDE GELS [J].
BARTLEY, TD ;
MURPHYHOLLAND, K ;
EVELEIGH, DE .
ANALYTICAL BIOCHEMISTRY, 1984, 140 (01) :157-161
[3]   SENSITIVE DETECTION OF ENDO-1,4-BETA-GLUCANASES AND ENDO-1,4-BETA-XYLANASES IN GELS [J].
BIELY, P ;
MARKOVIC, O ;
MISLOVICOVA, D .
ANALYTICAL BIOCHEMISTRY, 1985, 144 (01) :147-151
[4]  
BIELY P, 1988, BIOTECHNOL APPL BIOC, V10, P99
[5]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[6]  
CHEN C M, 1987, Bio-Technology (New York), V5, P274
[7]   ENZYMATIC-HYDROLYSIS OF CELLULOSE - IS THE CURRENT THEORY OF THE MECHANISMS OF HYDROLYSIS VALID [J].
ENARI, TM ;
NIKUPAAVOLA, ML .
CRC CRITICAL REVIEWS IN BIOTECHNOLOGY, 1987, 5 (01) :67-87
[8]   CELLULOLYTIC COMPLEX OF TRICHODERMA-REESEI QM-9414 - IMMUNOCHEMICAL APPROACH [J].
FAGERSTAM, LG ;
PETTERSSON, LG .
FEBS LETTERS, 1979, 98 (02) :363-367
[9]   SUBCELLULAR FRACTIONATION OF A HYPERCELLULOLYTIC MUTANT, TRICHODERMA-REESEI RUT-C30 - LOCALIZATION OF ENDOGLUCANASE IN MICROSOMAL FRACTION [J].
GLENN, M ;
GHOSH, A ;
GHOSH, BK .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1985, 50 (05) :1137-1143
[10]   CELLULASE FAMILIES AND THEIR GENES [J].
KNOWLES, J ;
LEHTOVAARA, P ;
TEERI, T .
TRENDS IN BIOTECHNOLOGY, 1987, 5 (09) :255-261