食品科学 ›› 2025, Vol. 46 ›› Issue (15): 155-155.doi: 10.7506/spkx1002-6630-20250224-115

• 生物工程 • 上一篇    下一篇

土曲霉阿魏酸酯酶异源表达、酶学性质及在阿魏酸制备中的应用

韩红祥,郭成城,魏胜华,陈阿娜,李松   

  1. (安徽工程大学生物与食品工程学院,安徽 芜湖 241000)
  • 出版日期:2025-08-15 发布日期:2025-07-22
  • 基金资助:
    安徽高校自然科学研究项目(KJ2021A0509);安徽省自然科学基金项目(2208085MC83)

Heterologous Expression and Enzymatic Characterization of Ferulic Esterase from Aspergillus terreus and Its Application in the Preparation of Ferulic Acid

HAN Hongxiang, GUO Chengcheng, WEI Shenghua, CHEN Ana, LI Song   

  1. (School of Biological and Food Engineering, Anhui Polytechnic University, Wuhu 241000, China)
  • Online:2025-08-15 Published:2025-07-22

摘要: 为提高阿魏酸酯酶(ferulic esterase,FAE)的发酵单位活力,将土曲霉FAE编码基因在毕赤酵母实现异源高效表达。重组毕赤酵母发酵活力为(17.38±0.34)U/mL,是原始菌株的35 倍左右。经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳检验得出,重组土曲霉阿魏酸酯酶(recombinant Aspergillus terreus ferulic esterase,rAtFAE)分子质量约为39 kDa。酶学性质研究表明rAtFAE最适pH值为6.0、最适反应温度为50 ℃,以咖啡酸甲酯、香豆酸甲酯、阿魏酸甲酯、芥子酸甲酯为底物时测得的kcat/Km值分别为255、237、112.64、96.85 L/(mol·min)。在以麸皮为原料制备阿魏酸的应用实验中,rAtFAE可以与木聚糖酶产生良好的协同作用并有效提升阿魏酸的释放量,最大释放量为(1 876.45±30.05)μg/g(以麸皮质量计)。

关键词: 土曲霉;阿魏酸酯酶;异源表达;毕赤酵母;阿魏酸

Abstract: To improve the expression level of ferulic esterase (FAE), the FAE encoding gene from Aspergillus terreus was heterologously expressed in Pichia pastoris. The expression level of FAE in the recombinant strain was (17.38 ± 0.34) U/mL, which was approximately 35 times as high as that of the original strain. The molecular mass of the recombinant A. terreus FAE (rAtFAE) was approximately 39 kDa as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Enzymatic characterization showed that the optimal pH of rAtFAE was 6.0, the optimal reaction temperature was 50 ℃, and the kcat/Km values measured with methyl caffeic acid, methyl coumarinate, methyl ferulic acid and methyl sinakoate as substrates were 255, 237, 112.64 and 96.85 L/(mol·min), respectively. The combination of rAtFAE and xylanase showed a good synergistic effect in enhancing the release of ferulic acid from wheat bran up to (1 876.45 ± 30.05) μg/g.

Key words: Aspergillus terreus; ferulic esterase; heterologous expression; Pichia pastoris; ferulic acid

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