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直接筛选酵母克隆文库以检测其对生淀粉水解的α-淀粉酶活性。

Direct screening of libraries of yeast clones for alpha-amylase activity on raw starch hydrolysis.

作者信息

Wong Dominic W S, Batt Sarah B, Lee Charles C, Robertson George H

机构信息

Western Regional Research Center, USDA-Agricultural Research Service, 800 Buchanan Street, Albany, CA 94710, USA.

出版信息

Protein Pept Lett. 2003 Oct;10(5):459-68. doi: 10.2174/0929866033478744.

Abstract

High-throughput screening for high-activity barley alpha-amylase mutants expressed in Saccharomyces cerevisiae is hampered by the interference of reducing agents, particularly the glucose used in yeast growth media. The present investigation employed colorimetric and chemiluminescent detection systems that enable direct and rapid screening of activities on raw starch substrate. Active clones could be separated into two groups, based on high total activity or high specific activity.

摘要

在酿酒酵母中表达的高活性大麦α-淀粉酶突变体的高通量筛选受到还原剂尤其是酵母生长培养基中使用的葡萄糖的干扰。本研究采用比色和化学发光检测系统,能够直接快速地筛选在生淀粉底物上的活性。根据总活性高或比活性高,活性克隆可分为两组。

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