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松茸的发酵条件优化及其对酪氨酸酶的抑制机理研究OA北大核心CSTPCD

Optimization of fermentation conditions of Tricholoma matsutake and mechanism of tyrosinase inhibition

中文摘要英文摘要

为了提高松茸的抗酪氨酸酶活性,利用嗜酸乳杆菌对其进行发酵,并探究其活性变化机理.通过单因素试验法优化了发酵条件,采用苯酚-硫酸法、二喹啉甲酸法和HPLC分析发酵前后物质含量变化,通过酶反应动力学分析、铜离子螯合实验和酪氨酸酶荧光猝灭实验研究发酵样品和酪氨酸酶之间的相互作用.结果表明,在嗜酸乳杆菌接种量5%,松茸添加量4%,发酵时间4 d时发酵样品有最大的酪氨酸酶抑制率,IC50值从发酵前的>10 mg/mL降低至(4.74±0.21)mg/mL.发酵提高了总蛋白和寡肽(<1 000 Da)的含量,促进了具有生物活性的寡肽和游离氨基酸的释放.进一步的机理研究表明,发酵样品通过混合型可逆抑制的方式降低酪氨酸酶活性,并能够螯合铜离子,半螯合浓度为(9.14±0.04)mg/mL,且能猝灭酪氨酸酶内源性荧光,说明发酵样品可能通过与铜离子结合,占据酪氨酸酶的活性位点,从而抑制其活性.嗜酸乳杆菌的发酵处理能显著提高松茸的酪氨酸酶抑制能力,研究结果为其在化妆品和功能性食品领域的应用提供了科学依据.

in this study,Tricholoma matsutake was fermented by using Lactobacillus acidophilus,to improve its tyrosinase inhibitory activity.The mechanism of activity change in Tricholoma matsutake after fermentation was also explored.The fermentation conditions were optimized by a single-factor experiment.The effects of fermentation on active ingredients were analyzed using the phenol-sulfuric acid meth-od,bicinchoninic acid method,and HPLC.Further,the interaction between fermented Tricholoma matsutake and tyrosinase was investiga-ted by using enzymatic reaction kinetics analysis,copper ion chelation assay,and tyrosinase fluorescence quenching assay.Results showed that the highest tyrosinase inhibition rate of fermented Tricholoma matsutake was observed at 5%inoculation amount of Lactobacillus aci-dophilus,4%of matsutake mushroom,and 4 days of fermentation time.And the IC50 value decreased from over 10 mg/mL to(4.74±0.21)mg/mL after fermentation.The content of total protein and oligopeptides(<1 000 Da)was increased after fermentation,suggesting that the fermentation improved the release of bioactive oligopeptides and free amino acids.Further mechanistic studies indicated that the fermented Tricholoma matsutake reduced tyrosinase activity by mixed reversible inhibition.In this process,it chelated copper ions at a semi-chelating concentration of(9.14±0.04)mg/mL and then quenched the endogenous fluorescence of tyrosinase.This indicated that the fermented Tricholoma matsutake may occupy the active site of tyrosinase by binding to copper ions,thereby inhibiting its activity.Re-sults showed that the tyrosinase inhibition ability of Tricholoma matsutake could be significantly improved after fermentation by Lactobacil-lus acidophilus.This work could lay a scientific foundation for its utilization in cosmetics and functional foods.

郭璟煊;刘学;刘春环;李颖;杨成

江南大学化学与材料工程学院,江苏无锡,214000国珍健康科技(北京)有限公司,北京,102200

发酵嗜酸乳杆菌松茸酪氨酸酶抑制机理探究

fermentationLactobacillus acidophilusTricholoma matsutaketyrosinasemechanism investigation

《食品与发酵工业》 2024 (013)

64-71 / 8

江苏省自然科学基金项目(BK20221069)

10.13995/j.cnki.11-1802/ts.039064

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