Characterization of a novel steviol-producing β-glucosidase from Penicillium decumbens and optimal production of the steviol
Characterization of a novel steviol-producing β-glucosidase from Penicillium decumbens and optimal production of the steviol
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DOI：
10.1007/s00253-013-4883-0
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发表时间：
2013-09-01
影响因子：
5
通讯作者：
Kim, Young-Min
中科院分区：
文献类型：
--
作者：
Ko, Jin-A;Ryu, Young Bae;Kim, Young-Min
本研究旨在开发一种经济可行的酶，用于从甜菊苷（ST）最佳生产甜菊醇（S）。从9种市售的葡萄糖苷酶中筛选出产S的β-葡萄糖苷酶（SPGase），并通过三步柱层析法从斜卧青霉柚苷酶中纯化了74倍。121-kDa蛋白在pH2.3 -6.0和40-60 ℃下稳定。通过SPGase水解ST产生甜茶苷（R）、甜菊双糖苷（SteB）、甜菊醇单葡糖苷（SMG）和S，如通过HPLC、HPLC-MS和H-1-和C-13-核磁共振所测定。SPGase对甜菊醇单葡糖基酯、ST、R和SMG显示出比其他β-连接的葡萄糖二糖更高的活性。S生产的最佳条件（30 mM，64%产率）是47 mM ST和43 μ l SPGase，pH 4.0和55 ℃。这是第一份报告详细介绍了生产S从ST水解的一种新的β-葡萄糖苷酶，这可能是有用的医药和农业领域。
This study aimed to develop an economically viable enzyme for the optimal production of steviol (S) from stevioside (ST). Of 9 commercially available glycosidases tested, S-producing beta-glucosidase (SPGase) was selected and purified 74-fold from Penicillium decumbens naringinase by a three-step column chromatography procedure. The 121-kDa protein was stable at pH 2.3-6.0 and at 40-60 A degrees C. Hydrolysis of ST by SPGase produced rubusoside (R), steviolbioside (SteB), steviol mono-glucoside (SMG), and S, as determined by HPLC, HPLC-MS, and H-1- and C-13-nuclear magnetic resonance. SPGase showed higher activity toward steviol mono-glucosyl ester, ST, R, and SMG than other beta-linked glucobioses. The optimal conditions for S production (30 mM, 64 % yield) were 47 mM ST and 43 mu l of SPGase at pH 4.0 and 55 A degrees C. This is the first report detailing the production of S from ST hydrolysis by a novel beta-glucosidase, which may be useful for the pharmaceutical and agricultural areas.