抗阿尔茨海默病的牡蛎内生真菌分离筛选及发酵条件优化
Isolation and Screening of Endophytic Fungi of Oyster Against Alzheimer's Disease and Optimization of Fermentation Conditions
李钰 1袁梦 1刘淑珍 1王凤舞 2仇宏伟3
作者信息
- 1. 青岛农业大学食品科学与工程学院,山东 青岛 266109
- 2. 青岛农业大学食品科学与工程学院,山东 青岛 266109;山东省特种食品技术创新中心,山东 青岛 266109
- 3. 青岛农业大学学报编辑部,山东 青岛 266109
- 折叠
摘要
以青岛海域野生牡蛎为原料,采用菌丝尖端切割法分离获得内生真菌,采用形态学和18S rDNA对其进行鉴定,并对其进行种属分类.结果表明从牡蛎中共分离得到13株真菌,经鉴定这13株真菌全部属于子囊菌门(Ascomycetes),其中7株为枝孢霉属(Cladosporium),是牡蛎内生真菌的优势菌属,占分离真菌总数的53%,另外青霉属(Penicillium)两株,节菱孢霉(Arthrinium)、蜡蚧菌属(Lecanicillium)、链格孢霉属(Alternaria)和黑团孢属(Periconia)的菌株各1株.对分离得到的牡蛎内生真菌进行体外抗阿尔茨海默病活性筛选,其中5株真菌发酵产物兼具抑制乙酰胆碱酯酶(AChE)和抑制β-淀粉样蛋白(Aβ1-42)聚集的活性.为进一步开发利用,采用单因素结合响应面方法对活性菌株 ML3发酵条件进行了优化.结果表明:最佳发酵条件为发酵温度28℃,发酵时间12天,种子液接种量10%.在此条件下,次生代谢产物 AChE抑制率和 Aβ1-42 相对抑制率分别为57.65%和52.43%,比未优化前分别提高了9.35%和7.63%,且影响上述两个指标的主次因素均为:种子液接种量＞发酵温度＞发酵时间.
Abstract
The endophytic fungi were isolated from wild oysters in Qingdao sea area by mycelial tip-cutting method,and identified by morphology and 18S rDNA,and categorized into species and genera.The re-sults showed that a total of 13 strains of fungi were isolated from oysters,and all of them were identified as belonging to Ascomycetes,among which 7 strains were Cladosporium,which is the dominant genus of oyster endophytic fungi and accounted for 53%of the total number of isolated fungi,in addition to two strains of Penicillium,and one strain each of Arthrinium,Lecanicillium,Alternaria,and Periconia.I-solated oyster endophytic fungi were screened for anti-Alzheimer's disease activity in vitro,and the fer-mentation products of five strains of the fungi possessed both acetylcholinesterase-inhibiting and β-amyloid(Aβ1-42)aggregation-inhibiting activities.For further development and utilization,the fermentation condi-tions of the ML3 strain were optimized by a one-way combined response surface methodology.The results showed that the optimal fermentation conditions were seed liquid inoculation of 10%at 28℃for 12 days.Under this condition,the inhibition rate of AChE and Aβ1-42 from secondary metabolites were 57.65%and 52.43%,respectively,which were 9.35%and 7.63%higher than those before the unoptimized,and the primary and secondary factors affecting the above two indexes were:seed liquor inoculation＞fermenta-tion temperature＞fermentation time.
关键词
牡蛎/内生真菌/阿尔茨海默病/发酵Key words
oyster/Endophytic fungi/Alzheimer's disease/fermentation引用本文复制引用
基金项目
山东省自然科学基金面上项目(ZR2020MB101)
出版年
2024
NETL
NSTL
万方数据