食品科学 ›› 0, Vol. ›› Issue (): 0-0.

• 生物工程 •    下一篇

一株产单宁酶嗜热真菌的鉴定、酶学性质分析及CAZymes表达研究

谷新晰,卢海强,刘亚娟,陈赛娟,谷子林,陈宝江   

  1. 河北农业大学
  • 收稿日期:2017-07-29 修回日期:2018-07-17 出版日期:2018-09-25 发布日期:2018-09-18
  • 通讯作者: 陈宝江 E-mail:chenbaojiang@vip.sina.com
  • 基金资助:
    嗜热甘露聚糖酶Loop特征及其对酶热稳定性作用的分子机制

Identification, properties and CAZymes expression analysis of the thermophilic tannase-producing fungus

2, 2,Sai-Juan CHEN 2,   

  • Received:2017-07-29 Revised:2018-07-17 Online:2018-09-25 Published:2018-09-18

摘要: 目的:对产单宁酶的1株嗜热真菌HBFH5进行鉴定,开展单宁酶酶学性质的分析及碳水化合物酶的转录组学研究,探究嗜热真菌HBHF5在食品酶制剂开发中的潜力。方法与结果:经对菌株的菌落、孢子形态观察及ITS序列比对分析,最终鉴定嗜热真菌HBHF5为烟曲霉(Aspergillus fumigatus ),命名为Aspergillus fumigatus HBHF5。经分析,菌株HBHF5在固态发酵培养时不产单宁酶,而液态诱导培养时,HBHF5菌株胞内和胞外均检测到单宁酶活性,且以胞外酶为主(94%),酶活力最高达136U/mL。单宁酶最适反应温度为60℃,在60℃处理30min,能够维持酶原活力的90%以上。该酶最适反应pH为6.0,在pH5.0-pH9.0范围内,能够维持60%以上的酶活力。不同金属离子对酶活力的影响存在差异,Cu2+、Fe3+、Mn2+和Zn2+等金属离子对该单宁酶活性抑制较强。经转录组学分析,该菌以麸皮为唯一碳源时,共有淀粉酶、纤维素酶和果胶酶等239个CAZymes酶类基因表达,其中糖苷水解酶类最为丰富,约占CAZymes酶类表达总数的70%。结论:A. fumigatus HBHF5是1株优良产酶菌株,具有食品酶制剂开发潜力的嗜热真菌。

关键词: 单宁酶, 嗜热酶, 脱涩, 转录组, 碳水化合物酶

Abstract: Purpose: In this study, identification the tannase-producing strains HBFH5, analysis of enzymatic properties and carbohydrate-degrading enzymes, the potential of the strain in food industry were investigated. Methods and results: Based on its morphology and ITS sequence alignment analysis, the strain HBHF5 was identified as Aspergillus fumigatus, named Aspergillus fumigatus HBHF5. The strain HBHF5 did not produce tannase under solid-state fermentation(SSF), and produced an extracellular tannase by submerged fermentation(SmF), the maximum (136 U/ml) tannase activity. The tannase activity was optimal at pH 6.0 and 60℃. For a treatment at 60℃ for 30 minute, samples could maintain more than 90% of the tannase activity, and showed nearly 60% of the maximal activity in the range of pH5.0 to pH9.0. The tannase activity was inhibited by some metal ions, such as Cu2+, Fe3+, Mn2+and Zn2+. The transcriptomics results showed that 239 CAZyme genes were expressed, such as amylase, cellulase and pectinase. The most group is the Glycoside hydrolase, accounting for about 70% of CAZymes expression. Result: It has also been demonstrated with the A. fumigatus HBHF5 strain that there is potential in the food-processing industry.

Key words: Tannase, Thermophilic enzyme, Astringent, Transcriptomics, Carbohydrate active enzymes

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