As the hydrolysis product of radish thioglucoside catalyzed by thioglucosidase, sulforaphene (SFE) is a kind of functional ingredient with broad market prospects in virtue of its strong anti-oxidation, anti cell mutation, anti-cancer, antibacterial and herbicidal activities. Due to the fact that free thioglucosidase can be only used once, researching immobilized thioglucosidase and its enzymatic properties has determinant impact on the practical preparation of SFE. However, current immobilized thioglucosidase unexceptionally suffer from serious limitations of low mechanical strength and decomposition by microorganisms, not enough to support the enzyme to achieve continuous industrial reaction. In this study, thioglucosidase was separated and purified from radish seeds by means of extraction, precipitation and membrane separation, so the specific activity of thioglucosidase was increased to 8 times of the original. Then the purified thioglucosidase was covalently immobilized on amine-based resin by glutaraldehyde and tests have been performed on the determination of optimal cross-linking time, optimal cross-linking pH and the molar ratio of free enzyme, glutaraldehyde and carrier amino groups. Therewith the immobilized thioglucosidase was proved to possess superior enzymatic properties including pH adaptation, temperature adaptation, thermal stability, reusability and storage stability, compared with the free thioglucosidase. After the optimal substrate concentration and flow rate was found, the yield of SFE produced by the immobilized thioglucosidase reactor column reached 9.10%, and the benefit of immobilized thioglucosidase is 2.91 times that of direct fermentation. The immobilized thioglucosidase on amine-based resin can be further implemented for the production of SFE through enzymatic method instead of chemical synthesis or direct fermentation.

作为硫代葡糖苷酶催化的萝卜硫代葡糖苷的水解产物，磺胺噻吩（SFE）具有很强的抗氧化，抗细胞突变，抗癌，抗菌和除草活性，是一种具有广阔的市场前景的功能性成分。由于游离硫代葡糖苷酶只能使用一次，因此研究固定化的硫代葡糖苷酶及其酶学性质对SFE的实际制备具有决定性的影响。然而，目前固定的硫葡糖苷酶毫无例外地受到低机械强度和微生物分解的严重限制，不足以支持该酶以实现连续的工业反应。在这项研究中，通过提取，沉淀和膜分离从萝卜种子中分离纯化出了硫代葡萄糖苷酶，因此，硫葡糖苷酶的比活性提高到原来的8倍。然后通过戊二醛将纯化的硫代葡萄糖苷酶共价固定在胺基树脂上，并进行了测试，以确定最佳的交联时间，最佳的交联pH以及游离酶，戊二醛与载体氨基的摩尔比。由此证明，与游离的硫葡糖苷酶相比，固定化的硫葡糖苷酶具有优异的酶学性质，包括pH适应性，温度适应性，热稳定性，可重复使用性和储存稳定性。找到最佳的底物浓度和流速后，固定的硫代葡萄糖苷酶反应器柱产生的SFE的收率达到9.10％，固定的硫代葡萄糖苷酶的效率是直接发酵的2.91倍。