通过 UDP-葡萄糖基转移酶催化的级联反应高效生物合成水杨梅苷。

IF 2 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biotechnology Letters Pub Date : 2024-04-01 Epub Date: 2024-01-06 DOI:10.1007/s10529-023-03453-0

Guosi Li, Qilin Xu, Nan Hu, Xinyang Liu, Yiqi Jiang, Hailong Xue, Yongjun Zang, Fucheng Zhu

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引用次数: 0

摘要

目的：水杨梅苷是一种重要的植物源芳香化合物，具有多种生物特性。本研究的主要目的是利用 UDP-葡萄糖基转移酶（UGT）和原位再生的 UDP-葡萄糖（UDPG）从酪醇中合成水杨梅苷：结果：拟南芥中的UDP-葡萄糖基转移酶85A1（UGT85A1）对酪醇具有高活性和区域选择性，被选中用于生产丹皮苷。然后，通过将 UGT85A1 与来自 Glycine max 的蔗糖合成酶（GmSuSy）耦合，构建了一个原位再生 UDPG 的体外级联反应。确定了最佳的 UGT85A1-GmSuSy 活性比为 1:2，以平衡盐基糖苷生成和 UDP 葡萄糖再生的效率。此外，还确定了不同的级联反应条件来生产丹皮苷。在优化条件下，在连续进料反应器中生产的皂苷滴度为 6.0 g/L，相应的摩尔转化率为 99.6%，比生产率为 199.1 mg/L/h：结论：这是迄今为止利用生物催化方法生产出的最高滴度的皂甙。

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Highly efficient biosynthesis of salidroside by a UDP-glucosyltransferase-catalyzed cascade reaction.

Objective: Salidroside is an important plant-derived aromatic compound with diverse biological properties. The main objective of this study was to synthesize salidroside from tyrosol using UDP-glucosyltransferase (UGT) with in situ regeneration of UDP-glucose (UDPG).

Results: The UDP-glucosyltransferase 85A1 (UGT85A1) from Arabidopsis thaliana, which showed high activity and regioselectivity towards tyrosol, was selected for the production of salidroside. Then, an in vitro cascade reaction for in situ regeneration of UDPG was constructed by coupling UGT85A1 to sucrose synthase from Glycine max (GmSuSy). The optimal UGT85A1-GmSuSy activity ratio of 1:2 was determined to balance the efficiency of salidroside production and UDP-glucose regeneration. Different cascade reaction conditions for salidroside production were also determined. Under the optimized condition, salidroside was produced at a titer of 6.0 g/L with a corresponding molar conversion of 99.6% and a specific productivity of 199.1 mg/L/h in a continuous feeding reactor.

Conclusion: This is the highest salidroside titer ever reported so far using biocatalytic approach.

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来源期刊

Biotechnology Letters 工程技术-生物工程与应用微生物

CiteScore

5.90

自引率

3.70%

发文量

108

审稿时长

1.2 months

期刊介绍： Biotechnology Letters is the world’s leading rapid-publication primary journal dedicated to biotechnology as a whole – that is to topics relating to actual or potential applications of biological reactions affected by microbial, plant or animal cells and biocatalysts derived from them. All relevant aspects of molecular biology, genetics and cell biochemistry, of process and reactor design, of pre- and post-treatment steps, and of manufacturing or service operations are therefore included. Contributions from industrial and academic laboratories are equally welcome. We also welcome contributions covering biotechnological aspects of regenerative medicine and biomaterials and also cancer biotechnology. Criteria for the acceptance of papers relate to our aim of publishing useful and informative results that will be of value to other workers in related fields. The emphasis is very much on novelty and immediacy in order to justify rapid publication of authors’ results. It should be noted, however, that we do not normally publish papers (but this is not absolute) that deal with unidentified consortia of microorganisms (e.g. as in activated sludge) as these results may not be easily reproducible in other laboratories. Papers describing the isolation and identification of microorganisms are not regarded as appropriate but such information can be appended as supporting information to a paper. Papers dealing with simple process development are usually considered to lack sufficient novelty or interest to warrant publication.