Mass production of rubusoside using a novel stevioside-specific β-glucosidase from Aspergillus aculeatus.

Jin-A Ko, Young-Min Kim, Young Bae Ryu, Hyung Jae Jeong, Tae-Soon Park, Su-Jin Park, Young-Jung Wee, Joong-Su Kim, Doman Kim, Woo Song Lee

文献索引：J. Agric. Food Chem. 60(24) , 6210-6, (2012)

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摘要

Rubusoside (R) is a natural sweetener and a solubilizing agent with antiangiogenic and antiallergic properties. However, currently, its production is quite expensive, and therefore, we have investigated nine commercially available glycosidases to optimize an economically viable R-production method. A stevioside (ST)-specific β-glucosidase (SSGase) was selected and purified 7-fold from Aspergillus aculeatus Viscozyme L by a two-step column chromatography procedure. The 79 kDa protein was stable from pH 3.0 to pH 7.0 at 50-60 °C. Hydrolysis of ST by SSGase produced R and steviol monoglucosyl ester as determined by (1)H and (13)C nuclear magnetic resonance (NMR). Importantly, SSGase showed higher activity toward ST than other β-linked glucobioses. The optimal conditions for R production were 280 mM ST and 16.6 μL of SSGase at pH 5.1 and 63 °C. This is the first discussion detailing the production of R by enzymatic hydrolysis of ST and is useful for the food additive and pharmaceutical industries.