Chassis engineering of Escherichia coli for trans‐4‐hydroxy‐l‐proline production,Microbial Biotechnology

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Chassis engineering of Escherichia coli for trans‐4‐hydroxy‐l‐proline production
Microbial Biotechnology ( IF 4.8 ) Pub Date : 2020-05-12 , DOI: 10.1111/1751-7915.13573
Xiulai Chen _{1,

2,

3} , Juyang Yi _{1,

4} , Wei Song _{1,

2,

3} , Jia Liu _{1,

2,

3} , Qiuling Luo _{1,

2,

3} , Liming Liu _{1,

2,

3}

Affiliation

Microbial production of trans‐4‐hydroxy‐l‐proline (Hyp) offers significant advantages over conventional chemical extraction. However, it is still challenging for industrial production of Hyp due to its low production efficiency. Here, chassis engineering was used for tailoring Escherichia coli cellular metabolism to enhance enzymatic production of Hyp. Specifically, four proline 4‐hydroxylases (P4H) were selected to convert l‐proline to Hyp, and the recombinant strain overexpressing DsP4H produced 32.5 g l⁻¹ Hyp with α‐ketoglutarate addition. To produce Hyp without α‐ketoglutarate addition, α‐ketoglutarate supply was enhanced by rewiring the TCA cycle and l‐proline degradation pathway, and oxygen transfer was improved by fine‐tuning heterologous haemoglobin expression. In a 5‐l fermenter, the engineered strain E. coliΔsucCDΔputA‐VHb_(L)‐DsP4H showed a significant increase in Hyp titre, conversion rate and productivity up to 49.8 g l⁻¹, 87.4% and 1.38 g l⁻¹ h⁻¹ respectively. This strategy described here provides an efficient method for production of Hyp, and it has a great potential in industrial application.

中文翻译：

大肠杆菌底盘工程技术，用于反式-4-羟基-1-脯氨酸的生产

与传统的化学提取方法相比，微生物产生的反式-4-羟基-1-脯氨酸（Hyp）具有明显的优势。但是，由于Hyp的生产效率低，因此对于工业生产Hyp仍然具有挑战性。在这里，底盘工程被用于定制大肠杆菌的细胞代谢，以增强Hyp的酶促生产。具体来说，选择了四个脯氨酸4-羟化酶（P4H）将l-脯氨酸转化为Hyp，过表达Ds P4H的重组菌株在添加α-酮戊二酸的情况下产生了32.5 g l ^-1 Hyp。要生产不添加α-酮戊二酸的Hyp，可通过重新连接TCA循环和l来增加α-酮戊二酸的供应。脯氨酸降解途径，以及通过微调异源血红蛋白的表达改善了氧的转移。在一个5升发酵罐中，该工程化菌株大肠杆菌Δ sucCD Δ PUTA -VHb _（L） - Ds的P4H显示在Hyp的滴度，转化率和生产率显著增加高达49.8克升^-1，87.4％和1.38克L- ^-分别为¹ h ^-1。本文介绍的这种策略为生产Hyp提供了一种有效的方法，在工业应用中具有巨大的潜力。

更新日期：2020-05-12

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