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使用工程化大肠杆菌和多酶级联的高成本有效的ursodeoxycholic酸全细胞生物合成

Xue Zhang^1,2, Jiagang Xin³, Mengyu Liu^1,2

¹School of Pharmaceutical Sciences and Food Engineering, Liaocheng University, Liaocheng, China.

Frontiers in microbiology

|February 11, 2025

概括

经过工程设计的大肠杆菌有效地从陈氧酸 (CDCA) 中合成乌尔索氧酸 (UDCA). 这种全细胞生物催化剂实现了高的UDCA转化率,为治疗肝脏和胆汁疾病提供了有前途的方法.

科学领域:

生物技术和代谢工程生物技术和代谢工程
酶催化和生物催化
制药合成制药合成

背景情况:

乌尔索多西胆酸 (UDCA) 是肝脏和胆道疾病的关键治疗剂.
目前的生物UDCA合成依赖于两步的酶化过程,使用陈二氧化醇酸 (CDCA) 作为基质.
现有的方法在UDCA生产的效率和可扩展性方面面临挑战.

研究的目的:

设计一种微生物菌株,从CDCA中直接进行UDCA的全细胞催化合成.
为了优化反应条件,提高UDCA生产.
为UDCA建立一个可扩展和高效的生物催化途径.

主要方法:

构建一个工程 * Escherichia coli * (UCA23) 菌株共表达四个关键酶:乳酸脱酶 (LDH),葡萄糖脱酶 (GDH),7α-基固醇脱酶 (7α-HSDH) 和7β-基固醇脱酶 (7β-HSDH).
优化全细胞催化反应参数,包括温度,pH值,有机溶剂和表面活性剂.
将UDCA合成过程扩展到2L水平.

主要成果:

工程UCA23菌株通过全细胞催化成功地从CDCA合成UDCA.
在优化的反应条件下,在2L尺度下,在2小时内实现了99%的转化率,即100毫米CDCA在2L尺度下在2小时内转化.
这代表了UDCA生物合成中高度CDCA基质报告的最高转化率.

结论:

开发的工程 *E. coli* 菌株和优化过程为UDCA生产提供了高效的生物催化方法.
这种全细胞方法克服了传统的多步骤酶合成的局限性.
这些发现为可扩展且具有成本效益的UDCA用于治疗应用的制造铺平了道路.

关键词:

7α-hydroxysteroid脱水酶的使用方法 7β-基类固醇脱酶.陈二氧化醇酸的使用方法葡萄糖脱水酶的使用乳酸脱酶的使用方法乌尔索多氧胆酸是什么意思整个细胞的催化作用.

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