Nguyen Thi Thanh Hanh, Seo Changseop, Gu Bon-Cheol, Lim Hee-Jung, Ha Jung-Min, Kim Seong-Bo, Park Jun-Seong, Kim Doman
Institute of Food Industrialization, Institutes of Green Bio Science & Technology, and Graduate School of International Agricultural Technology, Seoul National University, Pyeongchang-gun, Gangwon-do, 25354, South Korea.
CJ CheilJedang, Life Ingredient & Material Research Institute, Suwon, 16495, South Korea.
Biotechnol Lett. 2018 Jan;40(1):197-204. doi: 10.1007/s10529-017-2460-9. Epub 2017 Oct 24.
To purify and characterize a specific enzyme from a commercial pectinase for the production of steviol from stevioside (Ste) without adding organic solvent and to improve steviol production.
Commercial Sumizyme PX converted Ste to steviol with a yield of 98%. β-Glucosidase from Sumizyme PX (βglyPX) was purified in three steps with 12.5-fold purification and 51% yield. The specific activity of the purified βglyPX was 141 U/mg. The molecular weight of βglyPX was ~ 116 kDa on SDS-PAGE. Its optimum activity was at pH 3.5 and 65 °C. It was stable for 12 h up to 55 °C and for 24 h at pH 2-9.5. K values of βglyPX for pNPGal, oNPGlc, lactose, and Ste were 2.4, 0.7, 18, and 7.8 mM, respectively. The optimum conditions for steviol production were 55 °C, 900 U/ml, 80 mg Ste/ml, 12 h.
βglyPX contains great potential for industrial steviol production from Ste.
从市售果胶酶中纯化并鉴定一种特定酶，用于在不添加有机溶剂的情况下从甜菊糖苷（Ste）生产甜菊醇，并提高甜菊醇产量。
市售Sumizyme PX将Ste转化为甜菊醇，产率为98%。Sumizyme PX中的β-葡萄糖苷酶（βglyPX）经三步纯化，纯化倍数为12.5倍，产率为51%。纯化后的βglyPX比活性为141 U/mg。SDS-PAGE显示βglyPX的分子量约为116 kDa。其最佳活性在pH 3.5和65℃。在55℃下可稳定12小时，在pH 2 - 9.5下可稳定24小时。βglyPX对对硝基苯基半乳糖苷（pNPGal）、邻硝基苯基葡萄糖苷（oNPGlc）、乳糖和Ste的米氏常数（K）值分别为2.4、0.7、18和7.8 mM。生产甜菊醇的最佳条件为55℃、900 U/ml、80 mg Ste/ml、12小时。
βglyPX在从Ste工业化生产甜菊醇方面具有巨大潜力。
