关键词: 酿酒酵母, β-葡萄糖苷酶, 毕赤酵母, 高效表达, 2,6-二甲氧基对苯醌

Abstract: An engineered strain producing high β-glucosidase activity was constructed and applied in the production of 2,6-dimethoxybenzoquinone. The β-glucosidase gene was amplified from the genomic DNA of Saccharomyces cerevisiae CS1401 using PCR. The amplified gene was then ligated to pPICZαA vector and transformed to Pichia pastoris X33 for expression under the induction of methanol. The activity of the recombinant enzyme was measured, and S. cerevisiae CS1401 and the recombinant strain were separately cultured in wheat germ medium for the production of 2,6-dimethoxybenzoquinone. The results indicated that the β-glucosidase gene of S. cerevisiae CS1401 was efficiently expressed in the engineered strain of P. pastoris X33, giving a β-glucosidase activity of up to 4.5 U/mL, which was 5 times higher than that of S. cerevisiae CS1401. Besides, the yield of 2,6-dimethoxybenzoquinone produced by the recombinant strain in shaking flasks reached 935.7 μg/g, which was 40% higher than that of S. cerevisiae CS1401. Therefore, this engineered strain has a promising prospect of application in the microbial production of 2,6-dimethoxybenzoquinone.

Key words: Saccharomyces cerevisiae, β-glucosidase, Pichia pastoris, over-expression, 2,6-dimethoxybenzoquinone