冷藏石斑鱼特定腐败菌菌相高通量测序分析及蛋白降解菌筛选鉴定Screening and Identification of Specific Spoilage Bacteria in Chilled Grouper Based on High-throughput Sequencing and Protein Degradation Ability
黎铸毅,刘美娇,陈秋翰,杨学博,王敏杰,钟赛意,洪鹏志,朱春华,刘寿春
摘要(Abstract)：
【目的】研究珍珠龙胆石斑鱼（Epinephelus fuscoguttatus♀×Epinephelus lanceolatus♂）冷藏过程优势腐败菌的菌相变化，筛选鉴定具有强蛋白分解能力的特定腐败菌。【方法】通过质量指数法（QIM）评价、菌落计数、挥发性盐基氮（TVB-N）含量等指标判断鱼肉腐败进程；对新鲜（冷藏0 d）、次新鲜（冷藏4 d）和腐败（冷藏8 d）样品进行细菌16S V3-V4区域高通量测序，通过物种组成、Alpha和Beta多样性分析、物种韦恩图分析其菌群的结构、多样性和差异；通过传统培养法分离腐败样品菌株，酪蛋白琼脂筛选蛋白分解菌，对筛选出的菌进行革兰氏染色和16S rDNA测序鉴定。【结果】冷藏石斑鱼质量指数法的感官拒绝点为12 d，菌落总数（TVC）超标临界期为4 d,TVB-N超标临界期约为10 d；随冷藏时间延长，鱼肉表面菌群多样性下降，新鲜鱼肉、冷藏4 d、冷藏8 d鱼肉菌群结构在属水平上具有明显差异；新鲜鱼肉中菌群主要是不动杆菌属（Acinetobacter）、嗜冷杆菌属（Psychrobacter）、金黄杆菌属（Chryseobacterium），相对丰度分别为20.44%、15.93%、9.24%；冷藏4 d和8 d鱼肉菌群主要物种是希瓦氏菌属（Shewanella）、气单胞菌属（Aeromonas）、嗜冷菌属（Psychrobacter）、不动杆菌属（Acinetobacter），其相对丰度分别为32.27%、34.89%,30.28%、16.02%,12.90%、16.65%以及13.99%、5.67%；冷藏8 d时鱼肉中具有良好蛋白分解能力的菌株为气单胞菌属和希瓦氏菌属，其组成比例接近高通量测序结果。【结论】气单胞菌属和希瓦氏菌属是珍珠龙胆石斑鱼冷藏腐败过程的优势菌并具有较强的蛋白降解能力，本研究可为后续探究冷藏石斑鱼优势腐败菌的致腐能力和开发靶向抑菌保鲜技术提供基础。
关键词(KeyWords)： 石斑鱼;冷藏;腐败;高通量测序;蛋白降解菌
基金项目(Foundation): 广东海洋大学科研启动项目（060302042311）;; 广东省高等教育创新团队项目（2021KCXTD021）;; 茂名市罗非鱼优势特色产业集群项目（22282109-1）;; 广东省现代农业产业技术体系创新团队建设项目（2023KJ150）
作者(Author): 黎铸毅,刘美娇,陈秋翰,杨学博,王敏杰,钟赛意,洪鹏志,朱春华,刘寿春
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