Purification and Characterization of Myrosinase in Dolsan Leaf Mustard(Brassica juncea) and Changes in Myrosinase Activity during Fermentation of Leaf Mustard Kimchi

돌산갓의 Myrosinase 분리 정제 및 갓김치 숙성 중 Myrosinase 활성도의 변화

  • Published : 1994.06.30

Abstract

Myrosinase in leaf mustard was purified and characterized to furnish a grounding information for utilizing the pungent taste and the potential antimicrobial capability of Dolsan leaf mustard to enhance the taste and storage life of kimchi. When myrosinase was purified from leaf mustard through a series of DEAE Sephadex, chromatofocusing and Con A Sepharose column chromatography, specific activity of the enzyme increased 7107-fold compared with that of crude enzyme preparation, and 18.8% yield was obtained. The purified myrosinase showed the optimum pH of 5.9, isoelectric point of 4.6, molecular weight of 129 kD, Km of 0.206 mM, and Vmax of $2.039\;{\mu}M{\cdot}min^{-1}{\cdot}mg\;protein^{-1}$, respectively. The optimum concentration of L-ascorbic for the maximum activity of the enzyme was 0.6 mM, and the enzyme activity decreased at a higher concentration of L-ascorbic acid than 0.6 mM, showing almost no enzyme activity at a L-ascorbic acid concentration of higher than 2.0 mM. Myrosinase activity in leaf mustard kimchi immediately after the kimchi was formulated was shown to be about 70 nmol/min/mg protein which decreased rapidly after 3 days of storage at $20^{\circ}C$, showing that less than half and almost none of the enzyme activity was retained in 4 and 10 days of storage, respectively.

Keywords