油菜籽饼粕中硫苷的酶解条件优化及降解产物分析

doi:10.3864/j.issn.0578-1752.2014.02.018

Abstract

Abstract: 【Objective】The aim of this study was to figure out the optimal factors for directional conversion of glucosinolates to isothiocyanates in rapeseed meal degraded by endogenous and exogenous myrosinase and explore the impact of exogenous myrosinase on the components and content of glucosinolates degradation products. It will provide a basis for the comprehensive utilization of rapeseed meal. 【Method】 Myrosinase activity was evaluated in 4 kinds of buffer solutions to establish a better reaction system. The effects of different enzymolysis conditions included solid liquid ratio, hydrolysis time, reaction temperature, pH value, and amount of ascorbic acid on the conversion of glucosinolates in rapeseed meal to isothiocyanates by endogenous and exogenous myrosinase were investigated through single factors and orthogonal array design methods. The optimal condition for the conversion was obtained by analyzing the yield of isothiocyanates. Taking methylene chloride as the extraction agent, the degradation products were concentrated in rotary evaporator. The products from endogenous and exogenous myrosinase were detected by gas charomatographic mass spectrometry (GC-MS). The kind and content of compounds from the products were monitored. 【Result】 Results showed that among the four reaction systems, the myrosinase had the highest activity under citric acid-phosphate buffer and under the same conditions, exogenous myrosinase activity was significantly higher than that of the endogenous myrosinase. The content of isothiocyanates had no obvious increase when exogenous myrosinase was more than 20% of rapeseed meal, and it was good to add 20% exogenous myrosinase of rapeseed meal to rapeseed meal. The optimal enzymolysis conditions were as follows: ratio of solid to liquid was 1:15, pH was 6.0, reaction temperature was 50℃, amount of ascorbic acid 0.005 mg•g-1, the degradation time was 1h. Under this condition, the isothiocyanates content was 1.799 mg•g-1 which were 7.64 times than these by the autolysis in rapeseed meal. The isothiocyanates content generated by exogenous myrosinase was significantly higher than that by endogenous myrosinase, but both contents had the same trend with the reaction conditions. Seven compounds from exogenous myrosinase were identified by GC-MS from the enzymatic hydrolysates of glucosinolates and accounted for more than 67% of the extracted degradation products which included 1 kind of nitrile and 2 kinds of oxazolidinones and 4 kinds of isothiocyanates. Allyl isothiocyanate, 1-butene,4-isothiocyanate, cyclopentyl isothiocyanate and benzene, and 2-ethyl- isothiocyanate were obtained among glucosinolate degradation products. Besides, 1-butene,4-isothiocyanate had the highest content compared with the other compounds and accounted for 26.77% in the detected products. The nitrile was benzenepropanenitrile. Goitrin and 2,1-benzisoxazole were the two oxazolidinones. Four compounds were analyzed in degradation products from endogenous myrosinase and had lower content than the seven compounds detected in the products from exogenous myrosinase.【Conclusion】This study indicated the isothiocyanates content increased significantly by exogenous myrosinase. It can also be concluded from the experiments the optimal enzymolysis condition for glucosinolates and the main compounds of enzymatic hydrolysates of glucosinolates from rapeseed meal. 1-Butene,4-isothiocyanate had the highest content in the 4 kinds of isothiocyanates analyzed in the test.

Key words: rapeseed meal , glucosinolates , myrosinase , isothiocyanates , GC-MS

DING Yan-1, LI Li-Qian-1, CAO Rong-2, TANG Gen-Sheng-2, GU Zhen-Xin-1, HAN Yong-Bin-1. Effect of Enzymolysis Conditions on Glucosinolates in Rapeseed Meal and Identification of Their Degradation Products[J].Scientia Agricultura Sinica, 2014, 47(2): 383-393.

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