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目的研究在柚皮苷酶解液中加入聚山梨酯80后辅助酶解制备柚皮素。方法将聚山梨酯80加入柚皮苷酶解液中,以转化率为指标,通过单因素考察聚山梨酯80的质量浓度、pH值、温度、酶与底物的质量比、底物质量浓度及反应时间对转化率的影响,并通过正交试验优化其制备工艺;采用1H-NMR、13C-NMR鉴定水解产物。结果柚皮苷在最佳酶解条件下(聚山梨酯80质量浓度5 g/L、pH 4.0、温度37℃、酶与底物的质量比0.6、底物质量浓度30 mg/mL和反应时间12 h)的转化率为(97.4±1.2)%,远大于未加聚山梨酯80时柚皮苷的转化效率(56.7±1.4)%;反应产物相对分子质量为272.25,核磁共振谱证实产物为柚皮素。结论在柚皮苷酶解液中加入聚山梨酯80后酶解制备柚皮素,可以缩短反应时间、提高底物质量浓度、减少酶用量,从而显著提高转化效率。该工艺简单稳定,适合工业化生产。
Objective To study the preparation of naringenin by adding enzyme polysorbate 80 to naringin hydrolyzate. Methods Polysorbate 80 was added to naringin hydrolyzate. The conversion rate was taken as an index. The polysaccharide concentration, pH, temperature, mass ratio of enzyme and substrate, substrate concentration And reaction time on the conversion rate, and through orthogonal test to optimize its preparation process; 1H-NMR, 13C-NMR identification of hydrolyzate. Results Naringin had the best enzymatic hydrolysis conditions (polysorbate 80 mass concentration 5 g / L, pH 4.0, temperature 37 ℃, mass ratio of enzyme to substrate 0.6, substrate concentration 30 mg / mL and reaction time (97.4 ± 1.2)%, which was much higher than that of naringin (56.7 ± 1.4)% without polysorbate 80. The relative molecular mass of the reaction product was 272.25. The results of nuclear magnetic resonance spectroscopy showed that the product was Naringenin. Conclusion The addition of polysorbate 80 to the naringin enzymatic preparation of naringenin, can shorten the reaction time, increase the substrate concentration, reduce the amount of enzyme, which significantly improve the conversion efficiency. The process is simple and stable, suitable for industrial production.