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首页> 外文期刊>Journal of Agricultural and Food Chemistry >Development and Evaluation of an HPLC Method for Accurate Determinations of Enzyme Activities of Naringinase Complex
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Development and Evaluation of an HPLC Method for Accurate Determinations of Enzyme Activities of Naringinase Complex

机译：准确测定柚皮苷酶复合物酶活性的HPLC方法的开发和评估

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摘要

An HPLC method that can separate naringin, prunin, and naringenin was used to help accurately measure the activities of naringinase and its subunits (α-L-rhamnosidase and β-D-glucosidase). The activities of the naringinase and β-d-glucosidase were determined through an indirect calculation of the naringenin concentration to avoid interference from its poor solubility. The measured enzymatic activities of the naringinase complex, α-L-rhamnosidase, and β-D-glucosidase were the as same as their theoretical activities when the substrates' (i.e., naringin or prunin) concentrations were 200 μg/mL, and the enzyme concentrations were within the range of 0.06—0.43, 0.067—0.53, and 0.15—1.13 U/mL, respectively. The β-D-glucosidase had a much higher V_(max) than either naringinase or a-L-rhamnosidase, implying the hydrolysis of naringin to prunin was the limiting step of the enzyme reaction. The reliability of the method was finally validated through the repeatability test, indicating its feasibility for the determinations of the naringinase complex.
机译：可以分离出柚皮苷，李皮苷和柚皮苷的HPLC方法用于帮助准确测量柚皮苷酶及其亚基（α-L-鼠李糖苷酶和β-D-葡萄糖苷酶）的活性。通过间接计算柚皮苷浓度来确定柚皮苷酶和β-d-葡萄糖苷酶的活性，以避免其不良的溶解性。当底物浓度（即柚皮苷或李宁）浓度为200μg/ mL时，柚皮苷酶复合物，α-L-鼠李糖苷酶和β-D-葡糖苷酶的酶活性与理论活性相同。浓度分别在0.06-0.43、0.067-0.53和0.15-1.13 U / mL的范围内。 β-D-葡糖苷酶的V_（max）比柚皮苷酶或α-L-鼠李糖苷酶高得多，这表明柚皮苷水解为果胶是酶反应的限制步骤。最后通过重复性测试验证了该方法的可靠性，表明该方法可用于测定柚皮苷酶复合物。

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