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KMID : 0613820170270010038
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Journal of Life Science
2017 Volume.27 No. 1 p.38 ~ p.43
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Optimization of Glycosyl Aesculin Synthesis by Thermotoga neapolitana β-Glucosidase Using Response-surface Methodology
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Park Hyun-Su
Park Young-Don
Cha Jae-Ho
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Abstract
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Glycosyl aesculin, a potent anti-inflammatory agent, was synthesized by transglycosylation reaction, catalyzed by Thermotoga neapolitana β-glucosidase, with aesculin as an acceptor. The key reaction parameters were optimized using response-surface methodology (RSM) and 2 μg of the enzyme. As shown by a statistical analysis, a second-order polynomial model fitted well to the data (p<0.05). The response surface curve for the interaction between aesculin and other parameters revealed that the aesculin concentration and reaction time were the primary factors that affected the yield of glycosyl aesculin. Among the tested factors, the optimum values for glycosyl aesculin production were as follows: aesculin concentration of 9.5 g/l, temperature of 84℃, reaction time of 81 min, and pH of 8.2. Under these conditions, 61.7% of glycosyl aesculin was obtained, with a predicted yield of 5.86 g/l. The maximum amount of glycosyl aesculin was 6.02 g/l. Good agreement between the predicted and experimental results confirmed the validity of the RSM. The optimization of reaction conditions by RSM resulted in a 1.6-fold increase in the production of glycosyl aesculin as compared to the yield before optimization. These results indicate that RSM can be effectively used for process optimization in the synthesis of a variety of biologically active glycosides using bacterial glycosidases.
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KEYWORD
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Aesculin, β-glucosidase, response surface methodology, Thermotoga neapolitana, transglycosylation
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