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中国精品科技期刊2020

紫外联合酸胁迫红小豆萌发富集GABA工艺优化及其合成通路分析

Optimization of GABA Enrichment in Germinating Adzuki Bean under Combined Ultraviolet and Acid Stress and Analysis of Its Synthetic Pathway

  • 摘要: 为探究紫外联合酸胁迫对萌发红小豆中GABA富集的影响,通过单因素实验分析萌发时间、萌发温度、紫外胁迫时间及柠檬酸浓度对GABA合成的作用,结合响应面法优化萌发条件,并对相关代谢酶活性及其GABA合成通路进行分析。结果表明:紫外联合酸胁迫下红小豆萌发富集GABA的最佳工艺为:萌发时间73 h、萌发温度34 ℃、紫外胁迫时间4.5 h、柠檬酸浓度0.45 mmol/L,此条件下GABA含量最高,为159.83±1.32 mg/100 g(干基),是未萌发红小豆的6.32倍。酶活性测定显示,联合处理组的关键限速酶DAO酶活性在萌发24 h显著提升(P<0.05),GAD和PAO酶活性在48 h达到峰值,后期略有下降但仍维持催化能力,使GABA含量在73 h达到峰值,揭示酶活性动态调控在GABA积累中的作用。为进一步探究紫外联合酸胁迫红小豆萌发富集GABA的生物合成机制,利用转录组学进行分析,发现联合胁迫主要通过调控MAPK信号通路-植物、谷胱甘肽代谢、氮代谢等8条通路促进GABA合成。本研究证实紫外联合酸胁迫对萌发红小豆GABA富集具有显著促进作用,为解析其代谢调控机制及功能性红小豆加工提供理论参考。

     

    Abstract: This study aimed to investigate the effect of combined ultraviolet (UV) and acid stress on GABA enrichment in germinating adzuki beans (Vigna angularis). Single-factor experiments were conducted to analyze the impacts of germination time, temperature, UV stress duration, and citric acid concentration on GABA synthesis. Response surface methodology was applied to optimize the germination conditions, and the activities of related metabolic enzymes and the GABA synthesis pathway were analyzed. The results indicated that the optimal conditions for GABA enrichment in germinating adzuki beans under combined UV and acid stress was as follows: Germination time of 73 h, germination temperature of 34 ℃, UV stress duration of 4.5 h, and citric acid concentration of 0.45 mmol/L. Under these conditions, the maximum GABA content was 159.83±1.32 mg/100 g (dry weight), which was 6.32 times higher than that in non-germinating adzuki beans. Enzyme activity assays demonstrated that the activity of DAO, a key rate-limiting enzyme, was significantly increased at 24 h of germination in the combined treatment group. In comparison, the activities of GAD and PAO peaked at 48 h. Although their activities slightly decreased afterwards, the catalytic capacity was maintained, leading to the maximum GABA content at 73 h. These results revealed the role of dynamic regulation of enzyme activities in GABA accumulation. To further explore the biosynthetic mechanism of GABA enrichment in germinating adzuki beans under combined UV and acid stress, transcriptomic analysis was performed. The analysis revealed that this combined stress promoted GABA synthesis primarily by regulating eight pathways including MAPK signaling pathway-plant, glutathione metabolism, and nitrogen metabolism. This study confirmed that combined UV and acid stress could significantly promote GABA enrichment in germinating adzuki beans. It provides a theoretical reference for elucidating its metabolic regulation mechanism and the processing of functional adzuki bean products.

     

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