大肠杆菌全细胞催化产α-熊果苷的研究  

Production ofα-arbutin by Escherichia coli through whole-cell catalysis

作　　者：吴涵 毛银 周胜虎[1,2] 邓禹 WU Han;MAO Yin;ZHOU Shenghu;DENG Yu(National Engineering Research Center for Cereal Fermentation and Food Biomanufacturing,Jiangnan University,Wuxi 214122,China;School of Biotechnology,Jiangnan University,Wuxi 214122,China)

机构地区：[1]江南大学粮食发酵与食品生物制造国家工程研究中心,江苏无锡214122 [2]江南大学生物工程学院,江苏无锡214122

出　　处：《食品与发酵工业》2025年第4期131-138,共8页Food and Fermentation Industries

基　　金：国家重点研发计划项目(2022YFA0911800);江苏省杰出青年基金项目(BK20220089)。

摘　　要：α-熊果苷是二十一世纪理想的皮肤美白祛斑活性剂。当前,α-熊果苷的合成主要依赖酶催化或微生物发酵,由于产品价格昂贵,市场供不应求。因此,进一步提高α-熊果苷的产量有望降低生产成本和产品价格。该研究通过在大肠杆菌中异源表达肠膜明串珠菌的蔗糖磷酸化酶(sucrose phosphorylase,SPase),实现了全细胞催化产α-熊果苷的目标。随后,系统地分析了诱导温度、培养基类型、异丙基-β-D-硫代半乳糖苷(isopropyl-beta-D-thiogalactopyranoside,IPTG)浓度、诱导OD 600、接种量及诱导后培养时间对SPase的酶活性和表达水平的影响。在最佳产酶条件下全细胞催化显著提高了α-熊果苷的产量,总产量达到158.8 g/L,转化率为86.8%。为进一步提高全细胞催化次数,建立了固定化细胞技术并优化了催化过程参数,最终在14次循环催化后累计产量达到323.9 g/L,为α-熊果苷的工业化生产提供了技术参考。α-Arbutin is an ideal skin whitening and spot-fading active agent in the 21 st century.Currently,the synthesis ofα-arbutin mainly relies on enzyme catalysis or microbial fermentation,and due to the high product prices,there is a supply-demand imbalance in the market.Therefore,further increasing the yield ofα-arbutin is expected to reduce production costs and product prices.This study achieved the goal of whole-cell catalysis to produceα-arbutin by heterologously expressing the sucrose phosphorylase(SPase)from Leuconostoc mesenteroides in Escherichia coli.Subsequently,the effects of induction temperature,medium type,IPTG concentration,induction OD 600,inoculum size,and post-induction culture time on the enzymatic activity and expression level of SPase were systematically analyzed.Under optimal enzyme production conditions,whole-cell catalysis significantly increased the yield ofα-arbutin,with a total yield of 158.8 g/L and a conversion rate of 86.8%.To further increase the number of whole-cell catalysis cycles,immobilized cell technology was established,and the catalytic process parameters were optimized.Ultimately,a cumulative yield of 323.9 g/L was achieved after 14 cycles of catalysis,providing a technical reference for the industrial production ofα-arbutin.

关 键 词：α-熊果苷 蔗糖磷酸化酶 大肠杆菌 固定化细胞技术 对苯二酚 

分 类 号：TQ658[化学工程—精细化工] TQ203.2