具有酪氨酸酶抑制活性的乳酸菌筛选及抑制成分分析

摘要/Abstract

摘要： 酪氨酸酶在黑色素合成中起主要作用，并且已经进行了许多尝试来鉴定新的天然酪氨酸酶抑制剂，但很少有人试图在微生物中发挥作用。本研究以酪氨酸酶的抑制活性为筛选指标，评估了5株乳酸菌发酵上清液，菌体细胞和破碎提取物的酪氨酸酶抑制活性和DPPH自由基清除率，结果表明，发酵上清液抑制酪氨酸酶能力远高于菌体细胞和破碎提取物，其中MNJ-9菌株抑制酪氨酸酶能力可达68.9%。而菌体细胞和破碎提取物仅为50.9%和34.8%。MNJ-9菌株发酵上清液对DPPH自由基清除能力可达93.1%，因此对MNJ-9菌株进行16S rDNA菌种鉴定。基于液相色谱-质谱（Liquid Chromatography - Mass Spectrometry，LC-MS）非靶向代谢组学对MNJ-9菌株发酵上清液进行主成成分分析。对其主成成分前10种的化合物进行抑制率测定，结果表明排名第一的5-氨基戊酸抑制率最高。将5-氨基戊酸对小鼠黑色素瘤细胞（B16F10）进行细胞毒性实验、黑色素含量测定、细胞酪氨酸酶活性测定来评价其美白能力。结果表明，5-氨基戊酸浓度为70 μg/mL连续培养72 h时细胞存活率"≥" 80%，细胞无明显毒性。浓度为（10-70 μg/mL）时，细胞内黑色素含量和酪氨酸酶活性分别为（85.4%-45.2%）和（76.53%-40.5%），说明5-氨基戊酸具有一定的美白能力。

关键词: 乳酸菌, 酪氨酸酶抑制, DPPH自由基, 发酵上清液, 5-氨基戊酸, B16F10细胞

Abstract: Tyrosinase plays a major role in melanin synthesis and many attempts have been made to identify new natural tyrosinase inhibitors, but few attempts have been made to act in microorganisms. In this study, tyrosinase inhibition activity and DPPH free radical scavenging ratewas used as a screening indicator, the tyrosinase inhibitory activity and antioxidant capacity of five strains of Lactobacillus fermentum supernatant, bacteriophage cells and crushed extracts were evaluated, and the tyrosinase inhibitory capacity of fermentum supernatant was much higher than that of bacteriophage cells and crushed extracts, in which MNJ-9 Strain inhibited tyrosinase up to 68.9%. The fermentation supernatant of MNJ-9 Strain was able to scavenge DPPH free radicals up to 93.1%, Therefore, The 16S rDNA strain identification was performed on its MNJ-9 Strain. Principal component analysis of MNJ-9 Strain fermentation supernatant based on liquid chromatography - mass spectrometry (LC-MS) untargeted metabolomics. The inhibition rate was determined for the top 10 compounds of its main constituent，The results showed that 5-aminopentanoic acid。5-Aminopentanoic acid was subjected to cytotoxicity assay,melanin content assay and cellular tyrosinase activity assay on mouse melanoma cells (B16F10) to evaluate its whitening ability The results showed that 5-Aminopentanoic acid was incubated at 70 μg/mLwhen incubated continuously for 72 h. The cell survival rate was "≥" 80%, and the cells were not significantly toxic. The intracellular melanin content and tyrosinase activity at concentrations of (10-70 μg/mL) were (85.4%-45.2%) and (76.53%-40.5%) respectively, This indicates that 5-aminovaleric acid has some whitening ability.

Key words: Lactobacillus, Tyrosinase inhibition, DPPH free radicals, Fermentation supernatant, 5-Aminopentanoic acid, B16F10 cells

中图分类号:

Q939.11+7

张雪李雪峰胡瑞峰郭万春刘学军. 具有酪氨酸酶抑制活性的乳酸菌筛选及抑制成分分析[J]. 食品科学, 0, (): 0-0.

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