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OD600、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 of α-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 of α-arbutin is expected to reduce production costs and product prices.This study achieved the goal of whole-cell catalysis to produce α-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 OD600, 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 of α-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 of α-arbutin.
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