RAPID ISOLATION AND REGENERATION OF SCLEROTIUM-ROLFSII PROTOPLASTS AND THEIR POTENTIAL APPLICATION FOR STARCH HYDROLYSIS

被引：13

作者：

KELKAR, HS ^{[1
]}

SHANKAR, V ^{[1
]}

DESHPANDE, MV ^{[1
]}

机构：

[1] NATL CHEM LAB,DIV BIOCHEM SCI,POONA 411008,MAHARASHTRA,INDIA

来源：

ENZYME AND MICROBIAL TECHNOLOGY | 1990年 / 12卷 / 07期

关键词：

amylolytic enzymes; protoplast immobilization; Protoplast isolation; protoplast regeneration; S; rolfsii;

D O I：

10.1016/0141-0229(90)90067-Z

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：

A gentle and rapid procedure for obtaining high yield of Sclerotium rolfsii protoplasts is described. Optimum yields of protoplasts were obtained when 50 mg of 24-h-old mycelium was incubated with 5 mg of NovoZym 234 in 1 ml of 0.05 m maleic acid-NaOH buffer (pH 5.0) containing 0.6 m KCl. Isolated protoplasts were capable of high frequency regeneration (∼90-95%) when 0.6 m sucrose was used as an osmotic stabilizer. Since S. rolfsii protoplasts secrete a complete set of amylolytic enzymes, the isolated protoplasts were entrapped in calcium alginate gel and the immobilized system was tested for its ability to saccharify various starches. In batch operations, the immobilized preparation retained 57% of its initial activity after three cycles at 30°C and pH 5.0. © 1990.

引用

页码：510 / 514

页数：5

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