Engineering a Carbohydrate-processing Transglycosidase into Glycosyltransferase for Natural Product Glycodiversification.
Engineering a Carbohydrate-processing Transglycosidase into Glycosyltransferase for Natural Product Glycodiversification.
复制标题
将碳水化合物加工转糖苷酶改造为糖基转移酶，用于天然产物糖多样化。
DOI：
10.1038/srep21051
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发表时间：
2016-02-12
影响因子：
4.6
通讯作者：
Tang SY
中科院分区：
文献类型：
--
作者：
Liang C;Zhang Y;Jia Y;Wenzhao Wang;Li Y;Lu S;Jin JM;Tang SY
糖多样化拓宽了天然产物衍生药物发现的范围。利用蛋白质工程技术扩大了变形链球菌糖基转移酶D（GTF-D）的受体底物混杂性。在荧光底物4-甲基伞形酮上筛选位点饱和诱变文库中的突变体，以鉴定具有改善的转糖基化效率的衍生物。与野生型GTF-D酶相比，突变体M4在使用葡萄糖基供体蔗糖的情况下，对包括儿茶素、染料木素、大豆苷元和水飞蓟宾在内的类黄酮底物表现出增加的转糖基化能力。该研究证明了通过工程改造使用比NDP-糖便宜的供体底物的转糖苷酶来开发天然产物糖基转移酶的可行性，并且产生了一系列对于天然产物储库来说是新颖的α-葡萄糖基化天然产物。研究了染料木素α-葡萄糖苷的溶解性和儿茶素α-葡萄糖苷的抗氧化能力。
Glycodiversification broadens the scope of natural product-derived drug discovery. The acceptor substrate promiscuity of glucosyltransferase-D (GTF-D), a carbohydrate-processing enzyme fromStreptococcus mutans, was expanded by protein engineering. Mutants in a site-saturation mutagenesis library were screened on the fluorescent substrate 4-methylumbelliferone to identify derivatives with improved transglycosylation efficiency. In comparison to the wild-type GTF-D enzyme, mutant M4 exhibited increased transglycosylation capabilities on flavonoid substrates including catechin, genistein, daidzein and silybin, using the glucosyl donor sucrose. This study demonstrated the feasibility of developing natural product glycosyltransferases by engineering transglycosidases that use donor substrates cheaper than NDP-sugars, and gave rise to a series of α-glucosylated natural products that are novel to the natural product reservoir. The solubility of the α-glucoside of genistein and the anti-oxidant capability of the α-glucoside of catechin were also studied.