Molecular mechanism of high-production tannase of Aspergillus carbonarius NCUF M8 after ARTP mutagenesis: revealed by RNA-seq and molecular docking.

BACKGROUND: Tannase is an enzyme produced by microbial fermentation and is widely used in the food industry; however, the molecular mechanism of tannase production by Aspergillus has not yet been studied. This study was conducted to reveal the differences in Aspergillus carbonarius tannase enzymatic characterization, secondary structures and molecular mechanisms after treatment of the strain with atmospheric and room temperature plasma (ARTP). RESULTS: The results showed that the specific activity of tannase was improved by ARTP treatment, and it showed higher thermostability and tolerance to metal ions and additives. The enzymatic characterization and molecular docking results indicated that tannase had a higher affinity and catalytic rate with tannic acid as a substrate after ARTP treatment. In addition, the docking results indicated that Aspergillus tannases may catalyze tannic acid by forming two hydrogen-bonding networks with neighboring residues. RNA-seq analysis indicated that changes in steroid biosynthesis, glutathione metabolism, glycerolipid metabolism, oxidative phosphorylation pathway and mitogen-activated protein kinase signaling pathways might be crucial reasons for the high production of tannase. CONCLUSION: ARTP enhanced the yield and properties of A. carbonarius tannase by changing the enzyme structure and cell metabolism. This study provides a theoretical basis for elucidating the molecular mechanism underlying high production of Aspergillus tannases. © 2022 Society of Chemical Industry.

Physical modification on the in vitro digestibility of Tartary buckwheat starch: Repeated retrogradation under isothermal and non-isothermal conditions.

The effects of repeated retrogradation (RR, range from 1 to 3 times) at different temperatures (4 °C; 4/25 °C, with a 24 h interval; 25 °C) on the in vitro digestibility and structures of Tartary buckwheat starch (TS) were investigated in this study. Results demonstrated that TS treated by RR for 1 time under 4/25 °C contained the maximum content of slowly digestible starch (SDS, 35.25%); TS treated by RR for 3 times under 25 °C contained the maximum content of resistant starch (RS, 54.92%). As the increase of RR cycle times, the value of relative crystallinity, the ratios of 1047/1022 cm-1 and 995/1022 cm-1 increased, the starch pore wall thickened, and more smooth fragments appeared (observed by scanning electron microscope), while the value of melting temperature range trended to decrease. The crystallization type of TS changed from type "A" to a mixture of "B + V" after retrogradation treatment. Pearson correlation analysis revealed that the content of rapidly digestible starch (RDS) was negatively correlated with the ratio of 995/1022 cm-1, transition temperatures, and enthalpy (P < 0.05). These results would supply a potential method for the preparation of starch with slow-digesting properties, also improve the utilization and expand the application of TS.