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Colorimetric monitoring of the activity of recombinant Escherichia coli expressing styrene monooxygenase

机译:比色监测表达苯乙烯单加氧酶的重组大肠杆菌的活性

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摘要

Biocatalysis using recombinant Escherichia coli expressing styrene monooxygenase (SMO) activity is recognized as a promising process for the production of enantiopure aryl epoxides such as (S)-styrene oxide. Generally the activity of the whole cell biocatalyst is determined by measuring the conversion of styrene into (S)-styrene oxide by gas chromatography, which is however improper for the high-throughput analysis of many samples. Here we present that styrene monooxygenase catalyzed conversion of indole into indigo can be employed to monitor the activity of recombinant E. coli expressing styrene monooxygenase. We first confirmed that the colorimetric method was effective to monitor the whole cell activity of SMO by comparing the E. coli expressing styAB with that expressing only styB in a solid culture system. Next we used the monitoring method to investigate the effect of chaperone coexpression on the SMO whole cell activity in a liquid culture system.
机译:使用表达苯乙烯单加氧酶(SMO)活性的重组大肠杆菌进行生物催化被认为是生产对映体纯芳基环氧化物(如(S)-环氧乙烷)的有前途的过程。通常,整个细胞生物催化剂的活性是通过气相色谱法测量苯乙烯转化为(S)-环氧乙烷的转化率来确定的,但是,这不适用于许多样品的高通量分析。在这里,我们提出苯乙烯单加氧酶催化的吲哚向靛蓝的转化可以用来监测表达苯乙烯单加氧酶的重组大肠杆菌的活性。我们首先证实了比色法通过在固体培养系统中比较表达styAB的大肠杆菌和仅表达styB的大肠杆菌来有效监测SMO的整个细胞活性。接下来,我们使用监测方法来研究分子伴侣共表达对液体培养系统中SMO全细胞活性的影响。

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