Sulforaphane is a kind of isothiocyanate with antioxidant, anti-cancer and anticancer activities, but its application is limited due to its low yield. Myrosinase can hydrolyze glucosinolates to produce isothiocyanates. In this study, BoTGG1 from broccoli and BMYR from cabbage aphid were expressed in Pichia pastoris for the first time, respectively. The recombinant BMYR showed higher activity. And Pichia pastoris X33 combining with vector pPICZα were more conducive for the expression of myrosinase than GS115 and pPIC9K. High-cell density fermentation of the recombinant strain X33/pPICZα-BMYR was performed in a 3 L fermenter, and the activity of myrosinase reached 0.64 U/mL, which was 8-fold higher than that in shaking flask. Conversion of 3 g/L glucoraphanin using myrosi- nase at pH 4.5 and 45 ℃, 1.21 g/L of sulforaphane was obtained within 60 min, with the molar conversion rate of 99.6%, which is the highest level for the preparation of sulforaphane at present. This study provides a strategy for the production of myrosinase and preparation of sulforaphane.
HUANG Heou
,
ZHENG Pu
,
WU Dan
,
CHEN Pengcheng
. Heterologous expression of myrosinase and its application for preparation of sulforaphane[J]. Food and Fermentation Industries, 2022
, 48(22)
: 76
-81
.
DOI: 10.13995/j.cnki.11-1802/ts.031317
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