Efficient synthesis of β-lactam antibiotics with in situ product removal by a newly isolated penicillin G acylase.,Bioorganic Chemistry

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Efficient synthesis of β-lactam antibiotics with in situ product removal by a newly isolated penicillin G acylase.
Bioorganic Chemistry ( IF 4.5 ) Pub Date : 2020-03-16 , DOI: 10.1016/j.bioorg.2020.103765
Xin Pan ₁ , Anni Li ₂ , Zhiyi Peng ₃ , Xiaoqi Ji ₃ , Jianlin Chu ₂ , Bingfang He ₂

Affiliation

A penicillin G acylase (PGA) from Achromobacter xylosoxidans PX02 was newly isolated, and site-directed mutagenesis at three important positions αR141, αF142, βF24 was carried out for improving the enzymatic synthesis of β-lactam antibiotics. The efficient mutant βF24A was selected, and the (Ps/Ph)ini (ratio between the initial rate of synthesis and hydrolysis of the activated acyl donor) dramatically increased from 1.42-1.50 to 23.8-24.1 by means of the optimization of reaction conditions. Interestingly, the efficient enzymatic synthesis of ampicillin (99.1% conversion) and amoxicillin (98.7% conversion) from a high concentration (600 mM) of substrate 6-APA in the low acyl donor/nucleus ratio (1.1:1) resulted in a large amount of products precipitation from aqueous reaction solution. Meanwhile, the by-product D-phenylglycine was hardly precipitated, and 93.5% yield of precipitated ampicillin (561 mM) and 94.6% yield of precipitated amoxicillin (568 mM) were achieved with high purity (99%), which significantly simplified the downstream purification. This was the first study to achieve efficient β-lactam antibiotics synthesis process with in situ product removal, with barely any by-product formation. The effect enzymatic synthesis of antibiotics in aqueous reaction solution with in situ product removal provides a promising model for the industrial semi-synthesis of β-lactam antibiotics.

中文翻译：

通过新分离的青霉素G酰基转移酶原位去除产物，可高效合成β-内酰胺类抗生素。

从木氧化无色杆菌PX02中新分离出青霉素G酰基转移酶，并在三个重要位置αR141，αF142，βF24进行定点诱变，以改善β-内酰胺类抗生素的酶促合成。选择有效的突变体βF24A，并且通过优化反应条件，（Ps / Ph）ini（初始合成速率与活化的酰基供体的水解之间的比率）从1.42-1.50急剧增加至23.8-24.1。有趣的是，在低酰基供体/核比（1.1：1）下，由高浓度（600 mM）的底物6-APA有效地酶促合成氨苄青霉素（转化率为99.1％）和阿莫西林（转化率为98.7％）导致了一定数量的产物从反应水溶液中沉淀出来。与此同时，副产物D-苯基甘氨酸几乎不沉淀，高纯度（99％）的沉淀氨苄青霉素（561 mM）的产率为93.5％，阿莫西林沉淀的（568 mM）的产率为94.6％，显着简化了下游纯化。这是第一项通过原位去除产物而几乎不形成副产物的高效β-内酰胺类抗生素合成工艺的研究。反应水溶液中酶的酶促合成及原位产物去除效果为工业上半合成β-内酰胺类抗生素提供了一个有希望的模型。这是第一项通过原位去除产物而几乎不形成副产物的高效β-内酰胺类抗生素合成工艺的研究。反应水溶液中酶的酶促合成及原位产物去除效果为工业上半合成β-内酰胺类抗生素提供了一个有希望的模型。这是第一项通过原位去除产物而几乎不形成副产物的高效β-内酰胺类抗生素合成工艺的研究。反应水溶液中酶的酶促合成及原位产物去除效果为工业上半合成β-内酰胺类抗生素提供了有希望的模型。

更新日期：2020-04-20

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