Abstract
Tannase plays a crucial role in many fields, such as the pharmaceutical industry, beverage processing, and brewing. Although many tannases derived from bacteria and fungi have been thoroughly studied, those with good pH stabilities are still less reported. In this work, a mangrove-derived yeast strain Rhodosporidium diobovatum Q95, capable of efficiently degrading tannin, was screened to induce tannase, which exhibited an activity of up to 26.4 U/mL after 48 h cultivation in the presence of 15 g/L tannic acid. The tannase coding gene TANRD was cloned and expressed in Yarrowia lipolytica. The activity of recombinant tannase (named TanRd) was as high as 27.3 U/mL. TanRd was purified by chromatography and analysed by SDS-PAGE, showing a molecular weight of 75.1 kDa. The specific activity of TanRd towards tannic acid was 676.4 U/mg. Its highest activity was obtained at 40 °C, with more than 70% of the activity observed at 25–60 °C. Furthermore, it possessed at least 60% of the activity in a broad pH range of 2.5–6.5. Notably, TanRd was excellently stable at a pH range from 3.0 to 8.0; over 65% of its maximum activity remained after incubation. Besides, the broad substrate specificity of TanRd to esters of gallic acid has attracted wide attention. In view of the above, tannase resources were developed from mangrove-derived yeasts for the first time in this study. This tannase can become a promising material in tannin biodegradation and gallic acid production.
Highlights
Tannins are a class of poly-phenolic compounds widely existing in higher plants [1]
This tannase can become a promising material in tannin biodegradation and gallic acid production
HPLC analysis of the reaction mixture further verified that tannic acid can be converted gallic acid in the catalytic system of tannase generated by strain
Summary
Tannins are a class of poly-phenolic compounds widely existing in higher plants [1]. Among all the plant constituents, they rank the fourth in abundance, next to cellulose, hemicellulose, and lignin [2]. Considering the good thermal stability and the enzyme yield, tannases were mainly produced produced by by fungal fungal fermentation fermentation in industry, especially especially by by fermentation fermentation of of the the Aspergillus. Previous studies demonstrate that tannases from fromyeast yeast species beenverified only in verified in Sporidiobolus ruineniae, Candida sp., tannases species have have been only. Alongi et al found that yeasts were able to synthesize enzymes functioning in in the degradation of plant materials. We screened a tannin-degrading yeast strain degradation of plant materials [25,26]. Recombinant recombinant tannase featured excellentand pHdistinct stabilityrobustness, and distinct robustness, made potent tannase featured excellent pH stability which made itwhich a potent toolitina tannin tool in tannin biodegradation and gallic acid production.
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