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水稻糖基转移酶 OsUGT91C1 产生可口甜菊糖苷的催化灵活性。

Catalytic flexibility of rice glycosyltransferase OsUGT91C1 for the production of palatable steviol glycosides.

机构信息

Key Laboratory of Bio-Resource and Eco-Environment of Ministry of Education, College of Life Sciences, State Key Laboratory of Biotherapy and Cancer Center, West China Hospital of Sichuan University, Sichuan University, 610065, Chengdu, China.

Division of Structural Biology, Wellcome Trust Centre of Human Genomics, Roosevelt Drive, Oxford, OX3 7BN, UK.

出版信息

Nat Commun. 2021 Dec 2;12(1):7030. doi: 10.1038/s41467-021-27144-4.

DOI:10.1038/s41467-021-27144-4
PMID:34857750
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8639739/
Abstract

Steviol glycosides are the intensely sweet components of extracts from Stevia rebaudiana. These molecules comprise an invariant steviol aglycone decorated with variable glycans and could widely serve as a low-calorie sweetener. However, the most desirable steviol glycosides Reb D and Reb M, devoid of unpleasant aftertaste, are naturally produced only in trace amounts due to low levels of specific β (1-2) glucosylation in Stevia. Here, we report the biochemical and structural characterization of OsUGT91C1, a glycosyltransferase from Oryza sativa, which is efficient at catalyzing β (1-2) glucosylation. The enzyme's ability to bind steviol glycoside substrate in three modes underlies its flexibility to catalyze β (1-2) glucosylation in two distinct orientations as well as β (1-6) glucosylation. Guided by the structural insights, we engineer this enzyme to enhance the desirable β (1-2) glucosylation, eliminate β (1-6) glucosylation, and obtain a promising catalyst for the industrial production of naturally rare but palatable steviol glycosides.

摘要

甜菊醇糖苷是甜菊叶提取物中具有强烈甜味的成分。这些分子由一个不变的甜菊醇配基组成，上面装饰着可变的糖基，可广泛用作低热量甜味剂。然而，由于甜菊中特定的β（1-2）葡糖基化水平较低，最理想的甜菊醇糖苷 Reb D 和 Reb M（无不良余味）仅以痕量自然产生。在这里，我们报告了来自水稻的糖基转移酶 OsUGT91C1 的生化和结构特征，该酶在催化β（1-2）葡糖基化方面非常有效。该酶能够以三种模式结合甜菊醇糖苷底物，这使其能够灵活地以两种不同的取向以及β（1-6）葡糖基化催化β（1-2）葡糖基化。在结构见解的指导下，我们对该酶进行了工程改造，以增强所需的β（1-2）葡糖基化，消除β（1-6）葡糖基化，并获得了一种有前途的催化剂，可用于工业生产天然稀有但可口的甜菊醇糖苷。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bef0/8639739/66e266ba9e2e/41467_2021_27144_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bef0/8639739/6200827e193c/41467_2021_27144_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bef0/8639739/b7ae1daa529a/41467_2021_27144_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bef0/8639739/aa6b115b357a/41467_2021_27144_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bef0/8639739/d481da789ae3/41467_2021_27144_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bef0/8639739/66e266ba9e2e/41467_2021_27144_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bef0/8639739/6200827e193c/41467_2021_27144_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bef0/8639739/b7ae1daa529a/41467_2021_27144_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bef0/8639739/aa6b115b357a/41467_2021_27144_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bef0/8639739/d481da789ae3/41467_2021_27144_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bef0/8639739/66e266ba9e2e/41467_2021_27144_Fig5_HTML.jpg

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