食品与发酵工业 >

2023 , Vol. 49 >Issue 18: 224 - 231

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.034219

研究报告

柠檬苦素诱导塔宾曲霉UA13高产柠檬苦素转化酶的研究

  • 陈钰亭 ,
  • 李文杰 ,
  • 杨睿昕 ,
  • 雷生姣 ,
  • 王顺顺 ,
  • 杨丹丹 ,
  • 欧阳侬非
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  • 1(三峡大学 生物与制药学院,湖北 宜昌,443002)
    2(河南工业大学 国际教育学院,河南 郑州,450000)
第一作者:陈钰亭(硕士研究生)和李文杰(本科生)为共同第一作者(雷生姣副教授为通信作者,E-mail:leishengjiao@163.com)

收稿日期: 2022-11-05

  修回日期: 2022-12-04

  网络出版日期: 2023-10-25

基金资助

2022年湖北省柑橘产业链建设项目;宜昌海通食品有限公司(SDHZ2021293)

收起

Limonoid-induced high production of limonin invertase in Aspergillus tabin UA13

  • CHEN Yuting ,
  • LI Wenjie ,
  • YANG Ruixin ,
  • LEI Shengjiao ,
  • WANG Shunshun ,
  • YANG Dandan ,
  • OUYANG Nongfei
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  • 1(College of Biological and Pharmaceutical, China Three Gorges University, Yichang 443002, China)
    2(College of International Education, Henan University of Technology, Zhengzhou 450000, China)

Received date: 2022-11-05

  Revised date: 2022-12-04

  Online published: 2023-10-25

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摘要

以柠檬苦素为代表的三萜化合物是柑橘类水果在加工过程中产生“后苦味”的主要原因,采用酶法转化使柠檬苦素的含量降低是目前较适合果汁加工业的方法。该研究以实验室前期分离并经等离子体(atmospheric room temperature plasma,ARTP)诱变获得的塔宾曲霉UA13为出发菌株,采用柠檬苦素作为诱导物对其进行产酶诱导,并对菌株的发酵产酶工艺和酶的底物亲和性进行研究。结果表明,在1 L发酵罐培养基中,接种量10%(体积分数)、初始pH值5.0、搅拌转速250 r/min、30 ℃的最适条件下发酵,柠檬苦素转化酶活力最高可达48.52 U/mL。酶对柠檬苦素的底物亲和性研究结果表明,柠檬苦素转化酶的Km值为0.622 mmol/L,说明诱导后的菌株UA13所产酶对柠檬苦素的亲和力增强。同时研究结果显示,柠檬苦素转化酶具有较好的耐热性和耐酸性,在柑橘果汁加工业中具有非常好的应用前景。

关键词： 柠檬苦素; 诱导; 转化酶; 塔宾曲霉; 发酵罐优化; 柑橘果汁

本文引用格式

陈钰亭 , 李文杰 , 杨睿昕 , 雷生姣 , 王顺顺 , 杨丹丹 , 欧阳侬非 . 柠檬苦素诱导塔宾曲霉UA13高产柠檬苦素转化酶的研究[J]. 食品与发酵工业, 2023 , 49(18) : 224 -231 . DOI: 10.13995/j.cnki.11-1802/ts.034219

Abstract

Limonoid compounds existed in citrus are the major contributors to the bitter taste of citrus fruits. Limonin, the most representative limonoid compounds, produce an extraordinarily bitter taste after processing of fresh fruits without obvious bitter taste. The enzymatic method is suitable for removal of bitter components in citrus processed products nowadays. In this study, limonin is applied to induce the limonin invertase production in an Aspergillus tabin strain UA13, which was previously isolated in our laboratory and bred by atmospheric room temperature plasma (ARTP) mutagenesis. Furthermore, the fermentation and enzyme production were optimized and the substrate's affinity for the enzyme was investigated. The results demonstrated that the activity of limonin invertase by A. tabin UA13 reached up to 48.52 U/mL in 1 L fermentor at optimized conditions with 9.0 mg/L limonin induction, 10% (v/v) inoculum size, initial pH 5.0, 250 r/min stirring speed and 30 ℃ fermentation temperature. The results of limonin’s affinity for the enzyme displayed that the Km value of limonin converting enzyme was 0.622 mmol/L. This indicated that the enzyme-substrate affinity of limonin invertase produced by the A. tabin was significantly increased after limonin induction. Moreover, with high heat resistance and acid resistance, limonin invertase in this study is commercially potential and has prospective applications in the citrus processing industry for juice and concentrate.

Key words： limonin; induction; invertase; Aspergillus tabin; fermentation optimization; citrus juice

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