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预处理对绳状青霉纤维素酶水解纤维素及酶回收的影响。

Effect of pretreatment on the hydrolysis of cellulose by Penicillium funiculosum cellulase and recovery of enzyme.

作者信息

Rao M, Seeta R, Deshpande V

机构信息

Biochemistry Division, National Chemical Laboratory, Poona 411 008, India.

出版信息

Biotechnol Bioeng. 1983 Jul;25(7):1863-71. doi: 10.1002/bit.260250714.

Abstract

Penicillium funiculosum produces a complete cellulase which brings about 97% hydrolysis of cotton and has high beta-glucosidase, xylanase, laminarinase, and lichenase activities. This article deals with the effect of different pretreatments on the hydrolysis of sugarcane bagasse by P. funiculosum enzymes and the recovery of enzyme from the insoluble residues. Enzymic saccharification of bagasse pretreated with hot 1N NaOH followed by neutralization with HCI and steam treated under pressure (7 kg/cm(2)) gave 63 and 59% saccharification, respectively, in 48 h. Hemicellulose is not lost in these pretreatments. With a 30% slurry of steam-treated bagasse, a semisolid mass containing 14% sugar was obtained. A 90% recovery of CMCase, beta-glucosidase, and filter paper activity from the hydrolysates was obtained under the following conditions: (1) maintaining the ratio of enzyme to substrate high by stepwise addition of substrate, (2) brief grinding of the residual substrate with glass powder, and (3) addition of 0.4% Tween-80 to the eluting buffer. The high recovery of cellulolytic enzymes indicates that the adsorption of these enzymes on cellulose is not irreversible.

摘要

绳状青霉产生一种完整的纤维素酶,它能使棉花水解97%,并具有高β-葡萄糖苷酶、木聚糖酶、海带多糖酶和地衣多糖酶活性。本文探讨了不同预处理对绳状青霉酶水解甘蔗渣的影响以及从不溶性残渣中回收酶的情况。用热的1N氢氧化钠预处理甘蔗渣,然后用盐酸中和并在压力(7千克/平方厘米)下进行蒸汽处理,酶糖化在48小时内分别得到63%和59%的糖化率。在这些预处理中半纤维素不会损失。用30%的蒸汽处理甘蔗渣浆液可得到含糖量为14%的半固体物质。在以下条件下,从水解产物中可实现羧甲基纤维素酶、β-葡萄糖苷酶和滤纸酶活性90%的回收率:(1)通过逐步添加底物保持酶与底物的高比例,(2)用玻璃粉对残留底物进行短暂研磨,(3)向洗脱缓冲液中添加0.4%的吐温80。纤维素分解酶的高回收率表明这些酶在纤维素上的吸附不是不可逆的。

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