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利用嗜热栖热放线菌的重组耐热糖苷酶生物合成槲皮素。

Bioproduction of quercetin using recombinant thermostable glycosidases from Dictyoglomus thermophilum.

作者信息

Yu Shiqin, Shan Xiaoyu, Lyv Yunbin, Zhou Jingwen

机构信息

Science Center for Future Foods, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, Jiangsu, China.

Key Laboratory of Industrial Biotechnology, Ministry of Education and School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, Jiangsu, China.

出版信息

Bioresour Bioprocess. 2022 Apr 28;9(1):48. doi: 10.1186/s40643-022-00538-y.

Abstract

Quercetin is an essential ingredient in functional foods and nutritional supplements, as well as a promising therapeutic reagent. Also, the green technique to produce quercetin via rutin biotransformation is attractive. Genes encoding two thermostable glycosidases from Dictyoglomus thermophilum were cloned and expressed in Escherichia coli, which were applied in rutin biotransformation to produce highly pure quercetin at a high temperature. The production of biocatalysts were scaled up in a 5-L bioreactor, yielding a several-fold increase in total enzyme activity and a quercetin production of 14.22 ± 0.26 g/L from 30 g/L of rutin. Feeding strategies were optimized to boost biomass and enzyme production, achieving an activity of 104,801.80 ± 161.99 U/L for rhamnosidase and 12,637.23 ± 17.94 U/L for glucosidase, and a quercetin yield of 20.24 ± 0.27 g/L from the complete conversion of rutin. This study proposes a promising approach for producing high-quality quercetin in an industrial setting.

摘要

槲皮素是功能性食品和营养补充剂中的一种重要成分,也是一种有前景的治疗试剂。此外,通过芦丁生物转化生产槲皮素的绿色技术很有吸引力。从嗜热网柄菌中克隆了编码两种耐热糖苷酶的基因,并在大肠杆菌中表达,将其应用于芦丁生物转化以在高温下生产高纯度的槲皮素。在5-L生物反应器中扩大了生物催化剂的生产规模,使总酶活性提高了几倍,从30 g/L芦丁中生产出14.22±0.26 g/L的槲皮素。优化了补料策略以提高生物量和酶产量,鼠李糖苷酶活性达到104,801.80±161.99 U/L,葡萄糖苷酶活性达到12,637.23±17.94 U/L,芦丁完全转化后槲皮素产量为20.24±0.27 g/L。本研究提出了一种在工业环境中生产高质量槲皮素的有前景的方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/265e/10991118/769c53c8c7d4/40643_2022_538_Fig1_HTML.jpg

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