用于生物工艺开发的嗜酸氧化亚铁硫杆菌72W腈水解酶的蛋白质工程

Protein engineering of Acidovorax facilis 72W nitrilase for bioprocess development.

作者信息

Wu Shijun, Fogiel Arthur J, Petrillo Kelly L, Hann Eugenia C, Mersinger Lawrence J, DiCosimo Robert, O'Keefe Daniel P, Ben-Bassat Arie, Payne Mark S

机构信息

Central Research and Development Department, E.I. du Pont de Nemours and Co., Experimental Station, P.O. Box 80328, Wilmington, Delaware 19880-0328, USA.

出版信息

Biotechnol Bioeng. 2007 Jul 1;97(4):689-93. doi: 10.1002/bit.21289.

Abstract

Hydroxycarboxylic acid monomers can be used to prepare industrially important polymers. Enzymatic production of such hydroxycarboxylic acids is often preferred to chemical production since the reactions are run at ambient temperature, do not require strongly acidic or basic reaction conditions, and produce the desired product with high selectivity at high conversion. However, native enzymes often do not perform desired reactions with the efficiency required for commercial applications. Protein engineering was used to significantly increase the specific activity of nitrilase from Acidovorax facilis 72W for the conversion of 3-hydroxyvaleronitrile to 3-hydroxyvaleric acid. Overexpression of engineered nitrilase enzymes in Escherichia coli, combined with immobilization of whole cells in alginate beads that can be recycled many times has facilitated the development of a commercially viable bioprocess for production of 3-hydroxyvaleric acid.

摘要

羟基羧酸单体可用于制备具有重要工业价值的聚合物。此类羟基羧酸的酶促生产通常比化学合成更受青睐,因为反应在常温下进行,不需要强酸性或强碱性反应条件,并且能在高转化率下以高选择性生产所需产物。然而,天然酶往往无法以商业应用所需的效率进行预期反应。通过蛋白质工程显著提高了嗜酸栖热菌72W腈水解酶将3-羟基戊腈转化为3-羟基戊酸的比活性。在大肠杆菌中过表达工程腈水解酶,并将全细胞固定在可多次循环利用的藻酸盐珠中,这推动了一种用于生产3-羟基戊酸的具有商业可行性的生物工艺的发展。

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