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含酚羟化酶工程菌的两相体系细胞生物转化吲哚制备靛蓝。

Biotransformation of indole to indigo by the whole cells of phenol hydroxylase engineered strain in biphasic systems.

机构信息

State Key Laboratory of Urban Water Resource and Environment, Harbin Institute of Technology, Harbin, 150090, China.

出版信息

Appl Biochem Biotechnol. 2013 Feb;169(4):1088-97. doi: 10.1007/s12010-012-0069-y. Epub 2013 Jan 10.

Abstract

Biotransformation of indole to indigo in liquid-liquid biphasic systems was performed in Escherichia coli cells expressing phenol hydroxylase. It was suggested that indole could inhibit the cell growth even at low concentration of 0.1 g/L. The critical Log P for strain PH_(IND) was about 5.0. Three different solvents, i.e., decane, dodecane, and dioctyl phthalate, were selected as organic phase in biphasic media. The results showed that dodecane gave the highest yield of indigo (176.4 mg/L), which was more than that of single phase (90.5 mg/L). The optimal conditions for biotransformation evaluated by response surface methodology were as follows: 540.26 mg/L of indole concentration, 42.27 % of organic phase ratio, and 200 r/min of stirrer speed; under these conditions, the maximal production of indigo was 243.51 mg/L. This study proved that the potential application of strain PH_(IND) in the biotransformation of indole to indigo using liquid-liquid biphasic systems.

摘要

在表达苯酚羟化酶的大肠杆菌细胞中,在液-液两相体系中进行吲哚到靛蓝的生物转化。研究表明,即使在 0.1g/L 的低浓度下,吲哚也能抑制细胞生长。菌株 PH_(IND)的临界 Log P 约为 5.0。选择癸烷、十二烷和邻苯二甲酸二辛酯三种不同的溶剂作为两相中的有机相。结果表明,十二烷给出了最高的靛蓝产量(176.4mg/L),高于单相(90.5mg/L)。通过响应面法评估的生物转化最佳条件如下:吲哚浓度 540.26mg/L,有机相比例 42.27%,搅拌速度 200r/min;在此条件下,靛蓝的最大产量为 243.51mg/L。本研究证明了菌株 PH_(IND)在使用液-液两相体系将吲哚生物转化为靛蓝中的潜在应用。

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