Isolation, identification and fermentation process optimization of acetylcholinesterase inhibitory activity of endophytic fungus from Laminaria japonica
-
摘要: 本文以青岛海域的海带为原料,对其进行内生真菌的分离纯化,共得到7株内生真菌,采用改良Ellman法对其乙酰胆碱酯酶（ACh E）抑制活性进行筛选,并对其中一株活性最高内生真菌ML-X进行形态学和18S rDNA分子生物学鉴定,通过单因素实验和正交实验筛选出最适合该菌进行液体生长发酵的培养条件。结果表明,分离获得的菌株ML-X为白地霉属,序列号为KY977411,与遗传距离最近的Galactomyces geotrichum strain LMA-21株（JQ668739）同源性达99%。最佳发酵条件为:PS培养基,培养基的初始p H6.0,接种量5%,装瓶量150 m L/500 m L,培养温度31℃,发酵8 d,当发酵液粗浸膏为5 mg/m L时,对乙酰胆碱酯酶的抑制率可达28.91%±0.25%。Abstract: In this study, Laminaria japonica from Qingdao sea area was used to isolate and purify endophytic fungus.A total of seven strains of endophytic fungi were isolated. Using modified Ellman method to determine the inhibitory rate of ACh E and active strains were identified.Strain ML-X showed the highest AChE inhibitory activity and was identified, by physiologicalbiochemical characteristics and molecular biological techniques of 18 S rDNA sequencing.Based on single factor and orthogonal test, the culture conditions of liquid fermentation of the endophytic fungi were screened out. The results showed that the strain ML-X was confirmed as Geotrichum candidum, and the optimal fermentation conditions were as follows: PS medium was the suitable medium for the strain at pH6.0, incubated with 5% inoculum and 150 m L/500 m L culture volume at 31 ℃ for 8 days.Under these conditions, the strain ML-X exhibited a higher ACh E inhibitory activity, the inhibition ratio of Ach E was 28.91%± 0.25% when the concentration of crude extract from fermentation broth was 5 mg/m L.
-
[1] Snokusare S K, Kaul C L, Ramarao P.Demential of Alzheimer’s disease and other neurodegenerative disordersmemantine, a new hope pharmacological[J].Pharmaco-logical Research, 2005, 51 (1) :1-17.
[2] Terry R.D, Masliah E.Physical basis of cognitive alterationsin Alzheimer’s disease synapse loss is the major cor-relate of cognitive impairment[J].Ann Neurol, 1991, 30:572-580.
[3] 李前, 朱振霞.乙酰胆碱酯酶与老年性痴呆关系的研究进展[J].中国老年志, 2002, 4:325-326. [4] 秦云, 王凤舞.裙带菜内生菌的分离鉴定及其抑制ACh E活性的研究[J].现代食品科技, 2015, 31 (1) :53-58. [5] 王凤舞, 孟丽媛.1株具有抑制乙酰胆碱酯酶活性的牡蛎共生真菌的鉴定及发酵条件的优化[J].食品与发酵工业, 2014, 9:78-84. [6] 陈道金, 刘慧敏, 杨静, 等.植物内生菌活性成分研究进展[J].时珍国医国药, 2013, 24 (7) :1730-1732. [7] Strpbel G A.Endophytes as sources of bioactive products[J].Microbes Infect, 2003, 5 (6) :535-544.
[8] Wang F W, Ye Y H, Ding H, et al.Benzophenones from Guignardia sp.IFB-E028, an Endophyte on Hopea hainanensis[J].Chemistry&Biodiversity, 2010, 7 (1) :216-220.
[9] 赵宇, 张付云, 于清铭, 等.海带内生菌DNN6蛋白对小黄鱼保鲜作用的研究[J].食品工业科技, 2014, 35 (1) :321-325. [10] 李雪.海带内生菌DNN6代谢产物的分离纯化及其对小黄鱼保鲜的研究[D].沈阳:沈阳农业大学, 2016. [11] 赵宇.海带内生菌DNN7蛋白的分离及抗肿瘤活性研究[D].大连:大连海洋大学, 2015. [12] 赵宇, 刘浩舶, 李振, 等.海带内生菌HSN2胞外蛋白对粉红单端孢霉的影响[J].食品与机械, 2014, (6) :31-34, 114. [13] Ellman G L, Courtney K D, Featherstone RM, et al.Anew and rapid colorimetric determination of acetylcho-linesterase activity[J].Biochemical Pharmacology, 1961, 7 (2) :88-95.
[14] WANG Y, XU L, REN W.Bioactive metabolites from Chaetomium globosum L18, an endophytic fungus in themedicinal plant Curcuma wenyujin[J].Phytomedicine, 2012, 19 (3) :364-368.
[15] 孟丽媛.牡蛎共生真菌的分离鉴定及其抑制ACh E次生代谢产物的研究[J].青岛农业大学, 2013, 1:65-68. [16] 王广慧, 戴明, 魏雅冬.真菌液态发酵产漆酶的培养条件优化研究进展[J].食品工业, 2012, 33 (4) :130-134. -
期刊类型引用(3)
1. 胡智恺，索一平，李爽，王雨婷，刘薇，史锦硕，杨霞，姜洁. 婴儿配方奶粉中克罗诺杆菌的快速检测. 中国酿造. 2023(08): 253-259 . 
百度学术
2. 丁伯乐，蔡为荣，闻志莹，岳丹伟，朱樱，李晶晶. 山药低聚糖制备分离及对五种益生菌的增殖作用. 食品与发酵工业. 2020(24): 74-79 . 百度学术
3. 李婧，张柏林. 低聚葡萄糖对益生菌的增殖影响. 宁夏农林科技. 2019(12): 72-74+84 . 百度学术
其他类型引用(3)
计量
- 文章访问数: 180
- HTML全文浏览量: 22
- PDF下载量: 127
- 被引次数: 6
下载:
