Biosynthesis of 3-Hydroxyphloretin Using Rational Design of 4-Hydroxyphenylacetate 3-Monooxygenase,Journal of Agricultural and Food Chemistry

当前位置： X-MOL 学术 › J. Agric. Food Chem. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Biosynthesis of 3-Hydroxyphloretin Using Rational Design of 4-Hydroxyphenylacetate 3-Monooxygenase
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2023-11-29 , DOI: 10.1021/acs.jafc.3c06479
Shuping Xu ₁ , Pu Zheng ₁ , Ping Sun ₁ , Pengcheng Chen ₁ , Dan Wu ₁

Affiliation

The compound 3-hydroxyphloretin is a typical dihydrochalcone that can be obtained in plants by the 3-hydroxylation of phloretin. Here, the flavin-dependent two-component monooxygenase (HpaBC) derived from Pseudomonas aeruginosa was used to convert phloretin into 3-hydroxyphloretin. Following molecular docking and sequence alignment, modifications to the substrate pocket and loop of PaHpaBC were rationally designed, and mutant residues were selected. The results showed that the mutant Q212G/F292A/Q376N gave the best yield of 3-hydroxyphloretin and showed improved catalytic efficiency. Under optimal reaction condition, 2.03 g/L of 3-hydroxyphloretin was produced in the whole-cell catalysis experiment. Molecular docking and molecular dynamics simulations were used to analyze mutants and elucidate the potential mechanism. It was found that the increase in 3-hydroxyphloretin yield was due to the improvement in the flexibility of the loop and the expansion of its substrate pocket. This strategy based on loop and substrate pocket modification has significance in the engineering of PaHpaB.

中文翻译：

利用 4-羟基苯乙酸 3-单加氧酶的合理设计生物合成 3-羟基根皮素

化合物3-羟基根皮素是一种典型的二氢查耳酮，可通过根皮素的3-羟基化在植物中获得。在此，使用源自铜绿假单胞菌的黄素依赖性双组分单加氧酶（HpaBC）将根皮素转化为3-羟基根皮素。经过分子对接和序列比对，合理设计了Pa HpaBC底物口袋和环的修饰，并筛选出突变残基。结果表明，突变体Q212G/F292A/Q376N的3-羟基根皮素产率最高，催化效率提高。在最佳反应条件下，全细胞催化实验产生了2.03 g/L的3-羟基根皮素。使用分子对接和分子动力学模拟来分析突变体并阐明潜在机制。结果发现，3-羟基根皮素产量的增加是由于环的柔韧性的改善及其底物袋的扩张。这种基于环和底物袋修饰的策略在Pa HpaB 的工程中具有重要意义。

更新日期：2023-11-29

点击分享查看原文

点击收藏

阅读更多本刊新发论文本刊介绍/投稿指南