Microbial production of the plant flavanone hesperetin from caffeic acid.
Microbial production of the plant flavanone hesperetin from caffeic acid.
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DOI：
10.1186/s13104-023-06620-8
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发表时间：
2023-11-18
影响因子：
1.8
通讯作者：
Breitling, Rainer
中科院分区：
文献类型：
--
作者：
Hanko, Erik K. R.;Correia, Joao;Souza, Caio S.;Green, Alison;Chromy, Jakub;Stoney, Ruth;Yan, Cunyu;Takano, Eriko;Lousa, Diana;Soares, Claudio M.;Breitling, Rainer
关键词：
橙皮素是一种重要的O-甲基化类黄酮，由柑橘类水果产生，具有潜在的药用价值。橙皮素的微生物生物合成可能是植物提取的可行替代方案，因为植物提取物通常产生不同类黄酮的复杂混合物，使得分离纯化合物具有挑战性。在这项研究中，橙皮素生产的咖啡酸在微生物宿主大肠杆菌。我们结合了一个先前优化的途径，用于中间体黄烷酮圣草酚的生物合成与表达三个候选类黄酮O-甲基转移酶的质粒的组合文库。此外，我们努力提高CCoAOMT 7的位置特异性，CCoAOMT 7是一种来自拟南芥的类黄酮O-甲基转移酶，已被证明可以在帕拉和间位对圣草酚进行O-甲基化，从而产生橙皮素和高圣草酚的混合物。在我们的筛选中，发现表现最好的类黄酮O-甲基转移酶是CCoAOMT 7，其在E. coli 5-α。使用酶工程平台扫描选定关键位置的突变空间，使用同源性建模预测其结构，并使用对接模拟推断其潜在的催化改进，我们能够鉴定出对橙皮素具有两倍高位置特异性的CCoAOMT 7突变体。然而，突变体的催化活性大大降低。我们的研究结果表明，橙皮素可以从E.大肠杆菌中引入咖啡酸生物合成途径。在线版本包含补充材料，可通过10.1186/s13104-023-06620-8获得。
Hesperetin is an important O-methylated flavonoid produced by citrus fruits and of potential pharmaceutical relevance. The microbial biosynthesis of hesperetin could be a viable alternative to plant extraction, as plant extracts often yield complex mixtures of different flavonoids making it challenging to isolate pure compounds. In this study, hesperetin was produced from caffeic acid in the microbial host Escherichia coli. We combined a previously optimised pathway for the biosynthesis of the intermediate flavanone eriodictyol with a combinatorial library of plasmids expressing three candidate flavonoid O-methyltransferases. Moreover, we endeavoured to improve the position specificity of CCoAOMT7, a flavonoid O-methyltransferase from Arabidopsis thaliana that has been demonstrated to O-methylate eriodictyol in both the para- and meta-position, thus leading to a mixture of hesperetin and homoeriodictyol. The best performing flavonoid O-methyltransferase in our screen was found to be CCoAOMT7, which could produce up to 14.6 mg/L hesperetin and 3.8 mg/L homoeriodictyol from 3 mM caffeic acid in E. coli 5-alpha. Using a platform for enzyme engineering that scans the mutational space of selected key positions, predicting their structures using homology modelling and inferring their potential catalytic improvement using docking simulations, we were able to identify a CCoAOMT7 mutant with a two-fold higher position specificity for hesperetin. The mutant’s catalytic activity, however, was considerably diminished. Our findings suggest that hesperetin can be created from central carbon metabolism in E. coli following the introduction of a caffeic acid biosynthesis pathway. The online version contains supplementary material available at 10.1186/s13104-023-06620-8.
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影响因子：
64.8
作者：
Jumper J;Evans R;Pritzel A;Green T;Figurnov M;Ronneberger O;Tunyasuvunakool K;Bates R;Žídek A;Potapenko A;Bridgland A;Meyer C;Kohl SAA;Ballard AJ;Cowie A;Romera-Paredes B;Nikolov S;Jain R;Adler J;Back T;Petersen S;Reiman D;Clancy E;Zielinski M;Steinegger M;Pacholska M;Berghammer T;Bodenstein S;Silver D;Vinyals O;Senior AW;Kavukcuoglu K;Kohli P;Hassabis D
通讯作者：
Hassabis D
影响因子：
8.1
作者：
Ruviaro, Amanda Roggia;Menezes Barbosa, Paula de Paula;Macedo, Gabriela Alves
通讯作者：
Macedo, Gabriela Alves
影响因子：
3.5
作者：
Wils, Christopher Ralf;Brandt, Wolfgang;Vogt, Thomas
通讯作者：
Vogt, Thomas
影响因子：
5.8
作者：
Carbonell, Pablo;Wong, Jerry;Faulon, Jean-Loup
通讯作者：
Faulon, Jean-Loup
影响因子：
3
作者：
Trott, Oleg;Olson, Arthur J.
通讯作者：
Olson, Arthur J.