• 中国科技期刊卓越行动计划项目资助期刊
  • 中国精品科技期刊
  • 首都科技期刊卓越行动计划
  • EI
  • Scopus
  • CAB Abstracts
  • Global Health
  • 北大核心期刊
  • DOAJ
  • EBSCO
  • 中国核心学术期刊RCCSE A+
  • 中国科技核心期刊CSTPCD
  • JST China
  • FSTA
  • 中国农林核心期刊
  • 中国开放获取期刊数据库COAJ
  • CA
  • WJCI
  • 食品科学与工程领域高质量科技期刊分级目录第一方阵T1
中国精品科技期刊2020
杨淼,马志远,李延啸,等. 大曲中拜赖青霉发酵产单宁酶条件优化、纯化及酶学性质分析J. 食品工业科技,2026,47(8):1−9. doi: 10.13386/j.issn1002-0306.2025030409.
引用本文: 杨淼,马志远,李延啸,等. 大曲中拜赖青霉发酵产单宁酶条件优化、纯化及酶学性质分析J. 食品工业科技,2026,47(8):1−9. doi: 10.13386/j.issn1002-0306.2025030409.
YANG Miao, MA Zhiyuan, LI Yanxiao, et al. Optimization of Fermentation Conditions for Tannase Production by Penicillium bilaiae from Daqu, and Its Purification and Analysis of Enzymatic PropertiesJ. Science and Technology of Food Industry, 2026, 47(8): 1−9. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2025030409.
Citation: YANG Miao, MA Zhiyuan, LI Yanxiao, et al. Optimization of Fermentation Conditions for Tannase Production by Penicillium bilaiae from Daqu, and Its Purification and Analysis of Enzymatic PropertiesJ. Science and Technology of Food Industry, 2026, 47(8): 1−9. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2025030409.

大曲中拜赖青霉发酵产单宁酶条件优化、纯化及酶学性质分析

Optimization of Fermentation Conditions for Tannase Production by Penicillium bilaiae from Daqu, and Its Purification and Analysis of Enzymatic Properties

  • 摘要: 目的:为开发酶学性质优良的单宁酶资源。方法:通过透明圈平板筛选法初筛和甲醇罗丹宁比色法复筛,从大曲样品中筛选得到一株单宁酶活力最高的真菌,通过形态学观察和ITS序列测定进行菌种鉴定。优化了该菌株液体发酵产单宁酶的条件,通过硫酸铵沉淀和DEAE弱阴离子交换层析两步纯化,得到电泳级纯酶PbTanA。进一步探究PbTanA的酶学性质,最后评价了PbTanA在水解单宁酸生成没食子酸中的应用效果。结果:经鉴定,筛选出的菌株为拜赖青霉(Penicillium bilaise),命名为拜赖青霉CAU46-1。以3%单宁酸为诱导物,4%小麦麸皮为碳源,0.2%氯化铵为氮源,30 ℃、pH5.5条件下发酵4 d,发酵液单宁酶活力达28.7 U/mL,较优化前提高了3.3倍。PbTanA的分子量为79.0 kDa。该酶具有良好的耐酸性和热稳定性,最适pH为5.0,在pH2.0~7.0范围内稳定;最适温度为60 ℃,50 ℃下具有良好的温度稳定性。单宁酶PbTanA水解20 g/L单宁酸底物8 h,没食子酸浓度达18.4 g/L,得率为91.8%。结论:拜赖青霉CAU46-1所产的单宁酶PbTanA表现出优良的酶学特性,能够高效水解单宁酸生成没食子酸,具有良好的应用潜力。

     

    Abstract: Objective: To develop tannases resources with excellent enzymatic properties. Methods: Primary screening was conducted using the clear zone plate method, followed by a secondary screening using the methanol rhodanine colorimetric assay. A fungal strain with the highest tannase activity was isolated from the Daqu samples. The strain was identified through morphological observation and ITS sequence analysis. The liquid fermentation conditions for tannase production were optimized. The enzyme (PbTanA) was purified through ammonium sulfate precipitation and DEAE weak anion-exchange chromatography, obtaining electrophoretic-grade pure enzyme. The enzymatic properties of PbTanA were further investigated, and its application in hydrolyzing tannic acid to produce gallic acid was evaluated. Results: The strain was identified as Penicillium bilaiae and named Penicillium bilaiae CAU46-1. After fermentation at 30 ℃ and pH5.5 for 4 days, a tannase activity of 28.7 U/mL was achieved. This was achieved under conditions where 3% tannic acid served as inducer, 4% wheat bran as carbon source, and 0.2% ammonium chloride as nitrogen source, resulting in a tannase activity 3.3 times higher than the initial level. The tannase (PbTanA) was purified using ammonium sulfate precipitation followed by DEAE weak anion exchange chromatography. It was a molecular weight of 79.0 kDa. The enzyme exhibited good acid tolerance and thermal stability, with an optimal pH of 5.0 and good stability within the pH range of 2.0~7.0. Its optimal temperature was 60 ℃, and it remained stable up to 50 ℃. PbTanA hydrolyzed 20 g/L tannic acid for 8 h, producing a gallic acid concentration of 18.4 g/L with a yield of 91.8%. Conclusion: Tannase PbTanA from Penicillium bilaiae CAU46-1 showed excellent enzymatic properties and efficiently hydrolyzed tannic acid to produce gallic acid, demonstrating its potential for application.

     

/

返回文章
返回