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Quantification and identification of the main components of the Trichoderma cellulase complex with monoclonal antibodies using an enzyme-linked immunosorbent assay (ELISA).

作者信息

Kolbe J, Kubicek C P

机构信息

Abteilung für Mikrobielle Biochemie, Institut für Biochemische Technologie und Mikrobiologie, TU Wien, Vienna, Austria.

出版信息

Appl Microbiol Biotechnol. 1990 Oct;34(1):26-30. doi: 10.1007/BF00170918.

DOI:10.1007/BF00170918
PMID:1366972
Abstract

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 endoglucanase 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.

摘要

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Quantification and identification of the main components of the Trichoderma cellulase complex with monoclonal antibodies using an enzyme-linked immunosorbent assay (ELISA).
Appl Microbiol Biotechnol. 1990 Oct;34(1):26-30. doi: 10.1007/BF00170918.
2
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本文引用的文献

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Enzymatic hydrolysis of cellulose: is the current theory of the mechanisms of hydrolysis valid?纤维素的酶促水解:当前的水解机制理论是否有效?
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Immunoelectrophoretic detection of cellulases.纤维素酶的免疫电泳检测
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Recovery of functional proteins in sodium dodecyl sulfate gels.十二烷基硫酸钠凝胶中功能蛋白的回收
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Carbon Source Control of Cellobiohydrolase I and II Formation by Trichoderma reesei.里氏木霉细胞二糖水解酶 I 和 II 形成的碳源控制。
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8
Cellobiohydrolase II is the main conidial-bound cellulase in Trichoderma reesei and other Trichoderma strains.纤维二糖水解酶II是里氏木霉和其他木霉属菌株中主要的分生孢子结合型纤维素酶。
Arch Microbiol. 1991;155(6):601-6. doi: 10.1007/BF00245356.
9
Double-antibody sandwich enzyme-linked immunosorbent assay for quantitation of endoglucanase I of Trichoderma reesei.用于定量里氏木霉内切葡聚糖酶I的双抗体夹心酶联免疫吸附测定法。
Appl Environ Microbiol. 1991 Nov;57(11):3317-21. doi: 10.1128/aem.57.11.3317-3321.1991.
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4
A method for the detection and differentiation of cellulase components in polyacrylamide gels.一种在聚丙烯酰胺凝胶中检测和区分纤维素酶组分的方法。
Anal Biochem. 1984 Jul;140(1):157-61. doi: 10.1016/0003-2697(84)90147-7.
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Subcellular fractionation of a hypercellulolytic mutant, Trichoderma reesei Rut-C30: localization of endoglucanase in microsomal fraction.
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Sensitive detection of endo-1,4-beta-glucanases and endo-1,4-beta-xylanases in gels.凝胶中内切-1,4-β-葡聚糖酶和内切-1,4-β-木聚糖酶的灵敏检测
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Homologous domains in Trichoderma reesei cellulolytic enzymes: gene sequence and expression of cellobiohydrolase II.里氏木霉纤维素分解酶中的同源结构域:纤维二糖水解酶II的基因序列与表达
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EGIII, a new endoglucanase from Trichoderma reesei: the characterization of both gene and enzyme.来自里氏木霉的新型内切葡聚糖酶EGIII:基因与酶的特性研究
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Involvement of a conidial endoglucanase and a plasma-membrane-bound beta-glucosidase in the induction of endoglucanase synthesis by cellulose in Trichoderma reesei.分生孢子内切葡聚糖酶和质膜结合β-葡萄糖苷酶参与里氏木霉中纤维素诱导内切葡聚糖酶合成的过程。
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