MECHANISM OF ENZYMATIC CELLULOSE DEGRADATION - PURIFICATION OF A CELLULOLYTIC ENZYME FROM TRICHODERMA-VIRIDE ACTIVE ON HIGHLY ORDERED CELLULOSE
MECHANISM OF ENZYMATIC CELLULOSE DEGRADATION - PURIFICATION OF A CELLULOLYTIC ENZYME FROM TRICHODERMA-VIRIDE ACTIVE ON HIGHLY ORDERED CELLULOSE
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DOI：
10.1111/j.1432-1033.1973.tb02952.x
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发表时间：
1973-01-01
期刊：
影响因子：
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通讯作者：
PETTERSS.LG
中科院分区：
文献类型：
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作者：
BERGHEM, ER;PETTERSS.LG
< jats: p>< jats: list list-type=" explicit-label">< jats: list-item>< jats: p>纤维素分解酶（“C1< jats: sub>”酶）已从来源于真菌绿色木霉（Trichoderma viride）的培养物的商业纤维素酶制剂中分离< jats: italic>。< jats: list-item>< jats: p>纯化方法为四步程序，包括Bio‐Gel P‐10色谱、DEAE-Sephadex色谱、等电聚焦和Bio‐Gel P‐60色谱。< jats: list-item>< jats: p>每100克商品纤维素酶的产率为144毫克酶。< jats: list-item>< jats: p>分离的酶是均匀的聚丙烯酰胺凝胶电泳在pH 5.0和在pH 8.0的聚丙烯酰胺凝胶等电聚焦，也在ultracenthalge。< jats: list-item>< jats: p>在所用测定条件下，在纯化材料中未检测到对羧甲基纤维素的酶活性。同样，也没有β-葡萄糖苷酶活性。< jats: list-item>< jats: p>纯化的酶与3.3%的碳水化合物结合，并被认为是糖蛋白。该酶在pH3.79（10 ℃）下是等电的。通过在6 M盐酸胍中的Sepharose 6 B校准柱上对还原和烷基化酶进行色谱分析，测定分子量为46000。< jats: list-item>< jats: p>结晶纤维素（Avicel）、磷酸溶胀的Avicel和纤维四糖被酶降解，并且在每种情况下，主要反应产物是纤维二糖。< jats: list-item>< jats: p>有证据表明，纯化的酶是β-1，4-葡聚糖纤维二糖水解酶。
< jats: p>< jats: list list-type=" explicit-label">< jats: list-item>< jats: p> A cellulolytic enzyme (“C< jats: sub> 1” enzyme) has been isolated from a commercial cellulase preparation derived from culture filtrates of the fungus< jats: italic> Trichoderma viride.< jats: list-item>< jats: p> The purification method is a four‐step procedure including chromatography on Bio‐Gel P‐10, DEAE‐Sephadex chromatography, isoelectric focusing and chromatography on Bio‐Gel P‐60.< jats: list-item>< jats: p> A yield of 144 mg enzyme was obtained per 100 g commercial cellulase.< jats: list-item>< jats: p> The isolated enzyme was homogeneous in polyacrylamide gel electrophoresis at pH 5.0 and at pH 8.0 by isoelectric focusing in a polyacrylamide gel and also in the ultracentrifuge.< jats: list-item>< jats: p> No enzyme activity towards carboxymethylcellulose could be detected in the purified material under the assay conditions used. Similarly, there was no β‐glucosidase activity.< jats: list-item>< jats: p> The purified enzyme was associated with 3.3% carbohydrate and is assumed to be a glycoprotein. The enzyme was isoelectric at pH 3.79 (10 C). A molecular weight of 46000 was determined by chromatography of the reduced and alkylated enzyme on a calibrated column of Sepharose 6B in 6 M guanidine‐HCl.< jats: list-item>< jats: p> Crystalline cellulose (Avicel), phosphoric acid‐swollen Avicel and cellotetraose were degraded by the enzyme and in each case the principle reaction product was cellobiose.< jats: list-item>< jats: p> Evidence indicates that the purified enzyme is a β‐1, 4‐glucan cellobiohydrolase.