Hypericin is considered to be the most biologically active substance in the crude extract of Hypericum perforatum L. (also known as St. John's wort) and has a wide range of pharmacological effects. In this study, a high resolution high performance liquid chromatography method for determining hypericin was established by comparing different chromatographic conditions. Xylanase-assisted extraction and microwave-assisted extraction can improve the extraction yield of hypericin significantly. And the coupling strategy between two methods resulted in a significant difference on the extraction yield. Microwave-assisted extraction after xylanase-assisted extraction was found to be the most efficient strategy for extracting hypericin. The yield was 0.319 ± 0.006 mg g ⁻¹ , which was a 209.7% increase over unassisted extraction. The combination of enzyme assisted extraction followed by microwave assisted extraction can be more commonly applied to improve extraction efficiency of bioactive compounds from plants.

Glucosinolates can be hydrolyzed by the enzyme commonly known as myrosinase (E.C. 3.2.1.147)to a variety of biological compounds. Myrosinase (MYR)has been immobilized through the flexible spacers of different length on cross-linked chitosan resin (CCR). Ethylene diamine, hexamethylenediamine and decamethylene-diamine have been separately used as spacers. The influence of the flexible spacer length on the immobilized MYR (IMYR)properties were evaluated. The optimum pH and V _max of IMYR linearly increase with the flexible spacer length, and the optimum temperature and K _m of IMYR show an opposite trend. The recyclability of IMYR was good, with 90% recovery of activity after 10 cycles and 80% recovery after 30 cycles. IMYR was highly stable under storage conditions, with 95% recovery of activity after one year storage at 4 °C. The IMYR with the longest flexible spacer, decamethylene-diamine, was used as a biocatalyst for sulforaphene production. The overall hydrolysis ratio of glucoraphenin was 93.25 ± 0.91% and the activity of DDMCCR-IMYR remained 95% after 10 days of continuous use.