Ao Juwei, Pan Xuewei, Wang Qiang, Zhang Hengwei, Ren Kexin, Jiang An, Zhang Xian, Rao Zhiming
Key Laboratory of Industrial Biotechnology of Ministry of Education, Laboratory of Applied Microorganisms and Metabolic Engineering, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu 214122, P. R. China.
J Agric Food Chem. 2023 Feb 8;71(5):2438-2445. doi: 10.1021/acs.jafc.2c07972. Epub 2023 Jan 26.
α-Arbutin is extensively used in cosmetic industries. The lack of highly active enzymes and the cytotoxicity of hydroquinone limit the biosynthesis of α-arbutin. In this study, a whole-cell biocatalytic approach based on enzyme engineering and engineered cell modification was identified as effective in enhancing α-arbutin production. First, a sucrose phosphorylase (SPase) mutant with higher enzyme activity was obtained by experimental screening. Next, to avoid the oxidation of hydroquinone, we established an anaerobic process to improve the robustness of the cells by knocking out , , and in and overcoming the inhibitory effect of a high concentration of hydroquinone. Finally, the engineered strain was used for biotransformation in a 5 L fermenter with batch feeding for 24 h. The final yield of α-arbutin achieved was 129.6 g/L, which may provide a basis for the large-scale industrial production of α-arbutin.
α - 熊果苷在化妆品行业中被广泛使用。高活性酶的缺乏以及对苯二酚的细胞毒性限制了α - 熊果苷的生物合成。在本研究中，基于酶工程和工程细胞改造的全细胞生物催化方法被确定为有效提高α - 熊果苷产量的方法。首先，通过实验筛选获得了具有更高酶活性的蔗糖磷酸化酶（SPase）突变体。接下来，为避免对苯二酚的氧化，我们通过敲除大肠杆菌中的pgi、zwf和poxB并克服高浓度对苯二酚的抑制作用，建立了厌氧过程以提高细胞的稳健性。最后，将工程菌株用于5 L发酵罐中进行分批补料24 h的生物转化。α - 熊果苷的最终产量达到129.6 g/L，这可能为α - 熊果苷的大规模工业生产提供基础。
