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首页> 外文期刊>Journal of Biotechnology >Expression of a Dianthus flavonoid glucosyltransferase in Saccharomyces cerevisiae for whole-cell biocatalysis
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Expression of a Dianthus flavonoid glucosyltransferase in Saccharomyces cerevisiae for whole-cell biocatalysis

机译：石竹类黄酮葡糖基转移酶在酿酒酵母中的全细胞生物催化表达

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摘要

Glycosyltransferases are promising biocatalysts for the synthesis of small molecule glycosides. In this study, Saccharomyces cerevisiae expressing a flavonoid glucosyltransferase (GT) from Dianthus caryophyllus (carnation) was investigated as a whole-cell biocatalyst. Two yeast expression systems were compared using the flavonoid naringenin as a model substrate. Under in vitro conditions, naringenin-7-O-glucoside was formed and a higher specific glucosyl transfer activity was found using a galactose inducible expression system compared to a constitutive expression system. However, S. cerevisiae expressing the GT constitutively was significantly more productive than the galactose inducible system under in vivo conditions. Interestingly, the glycosides were recovered directly from the culture broth and did not accumulate intracellularly. A previously uncharacterized naringenin glycoside formed using the D. caryophyllus GT was identified as naringenin-4'-O-glucoside. It was found that S. cerevisiae cells hydrolyze naringenin-7-O-glucoside during whole-cell biocatalysis, resulting in a low final glycoside titer. When phloretin was added as a substrate to the yeast strain expressing the GT constitutively, the natural product phlorizin was formed. This study demonstrates S. cerevisiae is a promising whole-cell biocatalyst host for the production of valuable glycosides.
机译：糖基转移酶是用于合成小分子糖苷的有前途的生物催化剂。在这项研究中，研究了表达石竹（康乃馨）黄酮类葡糖基转移酶（GT）的酿酒酵母作为全细胞生物催化剂。使用类黄酮柚皮素作为模型底物，比较了两种酵母表达系统。在体外条件下，与组成型表达系统相比，形成了柚皮素-7-O-葡萄糖苷，并且使用半乳糖诱导型表达系统发现了更高的比葡萄糖基转移活性。但是，在体内条件下，组成型表达GT的酿酒酵母比半乳糖诱导系统的生产力要高得多。有趣的是，糖苷直接从培养液中回收，没有在细胞内积累。使用D. caryophyllus GT形成的先前未表征的柚皮苷糖苷被鉴定为柚皮苷4'-O-葡萄糖苷。发现酿酒酵母细胞在全细胞生物催化过程中水解柚皮素-7-O-葡糖苷，导致低的最终糖苷效价。当将芦丁作为底物添加到组成型表达GT的酵母菌株中时，形成了天然产物phlorizin。这项研究表明酿酒酵母是一种有前途的全细胞生物催化剂宿主，可用于生产有价值的糖苷。

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