%0 Journal Article %A GAO Zhaojian %A XU Xiang %A WANG Xianfeng %A LU Ning %A WANG Xudong %A ZHANG Tiezhu %A ZHANG Kangzhen %A JIAO Wei %T Enzymatic Properties and Application in Biomass Conversion of Acidophilic Feruloyl Esterase from Trichoderma atroatroviride %D 2019 %R 10.7506/spkx1002-6630-20180522-311 %J FOOD SCIENCE %P 121-128 %V 40 %N 10 %X In order to realize efficient biomass degradation in the preparation of ferulic acid, exerting important physiological functions in the body, feruloyl esterase (TaFAE) was isolated and purified from the fermentation broth of Trichoderma atroatroviride XS6 and its enzymatic characteristics were described in this experiment. The electrophoretically pure enzyme was obtained by consecutive ammonium sulfate precipitation, DEAE-cellulose DE52 anion exchange chromatography and Sephadex G-200 gel filtration chromatography. SDS-polyacrylamide gel electrophoresis (SDS-PAGE) of the enzyme revealed a single band of about 66.4 kDa. Its specific activity was 134.3 U/mg, with a recovery of 28.9%, and 31.52-fold purification. TaFAE had the highest affinity for methyl ferulate. When methyl ferulate was used as substrate, the Km and Vmax values of the enzyme were 0.53 mmol/L and 16.74 μmol/(min·mg), respectively. When methyl sultacide was used as substrate, the highest enzymatic reaction rate of 32.21 μmol/(min·mg) was obtained, which was twice as high as that of methyl ferulate. In addition, no activity was shown toward methyl caffeate, indicating that the enzyme has a stringent substrate specificity. The optimum pH and temperature of TaFAE were pH 4.0 and 40 ℃, respectively. In the pH range of 2.0–9.0 and at 30–40 ℃, TaFAE showed good stability. The metal ions Ca2+ and Mg2+ promoted TaFAE activity, whereas Hg2+ and Pb2+ almost completely inhibited the activity of the enzyme; β-mercaptoethanol, dithiothreitol (DTT), SDS, and Trition X-100 promoted the enzymatic activity, whereas PMSF had a strong inhibitory effect. Biomass conversion by xylanase produced significantly more ferulic acid and reducing sugar when TaFAE was added to the system, indicating that TaFAE and xylanase have a good synergistic effect. In summary, TaFAE has good acid tolerance and enzymatic properties, which make it promising for applications in the feed and food industry. %U https://www.spkx.net.cn/EN/10.7506/spkx1002-6630-20180522-311