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对黑曲霉进行高内涵筛选,以提高葡萄糖氧化酶的产量和分泌率。

High-content screening of Aspergillus niger with both increased production and high secretion rate of glucose oxidase.

作者信息

Zhu Xudong, Sun Jingchun, Chu Ju

机构信息

State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, P.O. Box 329, Shanghai, 200237, People's Republic of China.

出版信息

Biotechnol Lett. 2018 Jan;40(1):103-110. doi: 10.1007/s10529-017-2442-y. Epub 2017 Sep 22.

Abstract

OBJECTIVES

To develop a rapid, dual-parameter, plate-based screening process to improve production and secretion rate of glucose oxidase simultaneously in Aspergillus niger.

RESULTS

A morphology engineering based on CaCO was implemented, where the yield of GOD by A. niger was increased by up to 50%. Analysis of extracellular GOD activity was achieved in 96-well plates. There was a close negative correlation between the total GOD activity and its residual glucose of the fermentation broth. Based on this, a rapid, plate-based, qualitative analysis method of the total GOD activity was developed. Compared with the conventional analysis method using o-dianisidine, a correlation coefficient of -0.92 by statistical analysis was obtained.

CONCLUSION

Using this dual-parameter screening method, we acquired a strain with GOD activity of 3126 U l, which was 146% higher than the original strain. Its secretion rate of GOD was 83, 32% higher than the original strain.

摘要

目的

开发一种快速、基于平板的双参数筛选方法,以同时提高黑曲霉中葡萄糖氧化酶的产量和分泌率。

结果

实施了基于碳酸钙的形态工程,黑曲霉产生的葡萄糖氧化酶产量提高了50%。在96孔板中实现了细胞外葡萄糖氧化酶活性的分析。发酵液中葡萄糖氧化酶的总活性与其残留葡萄糖之间存在密切的负相关。基于此,开发了一种快速、基于平板的葡萄糖氧化酶总活性定性分析方法。与使用邻联茴香胺的传统分析方法相比,经统计分析得到的相关系数为-0.92。

结论

使用这种双参数筛选方法,我们获得了一株葡萄糖氧化酶活性为3126 U/l的菌株,比原始菌株高146%。其葡萄糖氧化酶分泌率为83,比原始菌株高32%。

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