ZHANG Nan,LI Yong,XU Jie,et al.Medium Optimization for Cordycepin Production From Cordyceps militaris by Submerged Fermentation[J].Northern Horticulture,2017,41(05):134-141.[doi:10.11937/bfyy.201705032]
蛹虫草深层发酵产虫草素培养基的优化
- Title:
- Medium Optimization for Cordycepin Production From Cordyceps militaris by Submerged Fermentation
- Keywords:
- Cordyceps militari; submerged fermentation; cordycepin; mycelial growth; medium; optimization
- 文献标志码:
- A
- 摘要:
- 以蛹虫草(Cordyceps militaris (L.) Link)NS-810为菌种，通过对接种量的考察，探索不同孢子浓度对蛹虫草液体一级种制作效果的影响；通过单次单因子试验和正交实验，优化深层发酵产虫草素的最佳培养基，筛选制备蛹虫草液体一级种和液体发酵生产虫草素的最佳工艺。结果表明：孢子浓度3.0×108 cfu?mL-1时制作的母种最适合作为蛹虫草菌种扩大培养中的一级种子液；深层发酵的最佳培养基配方为葡萄糖25 g?L-1、土豆100 g?L-1、鱼蛋白胨18 g?L-1、(NH4)2SO4 0.8 g?L-1、KH2PO4 1.0 g?L-1、MgSO4?7H2O 0.5 g?L-1、蚕蛹粉5.0 g?L-1、维生素B1 18 mg?L-1、水1 L。优化后虫草素的总产量为1 144.31 mg?L-1，较基础培养基提高了1.46倍。分别以价格低廉葡萄糖和鱼蛋白胨作为发酵培养基的碳源和有机氮源，利于蛹虫草产业化发酵生产虫草素。
- Abstract:
- In order to explore the optimal preparation method of liquid seed and medium for cordycepin production in submerged cultivation of Cordyceps militaris (L.) Link NS-810，effect of liquid seed on mycelial growth by inoculating different concentrations of spores in seed culture was investigated.Single factor experiment and orthogonal experiment were used to select the optimal medium compositions for cordycepin production in liquid fermentation.The results showed that the best concentration of spores for first-degree spawn was 3.0×108 cfu?mL-1.Under this condition，the pale yellow hypha ball dispersed homogeneously in the seed liquid and their diameters were in the range of 1.1 mm to 1.3 mm，and the seed liquid was clear and transparent.Furthermore，the optimal media was composed of the following factors:glucose 25 g?L-1，potato 100 g?L-1，peptone (fish) 18 g?L-1，(NH4)2SO4 0.8 g?L-1，KH2PO4 1.0 g?L-1，MgSO4?7H2O 0.5 g?L-1，silkworm pupa powder 5.0 g?L-1，water 1 L.This optimization strategy enhanced the production of cordycepin nearly 1.46 times compared with the initial level.Low-cost glucose and peptone (fish) were used as quick-release carbon source and organic nitrogen source of fermentation medium respectively，which was beneficial for industrial production of cordycepin from the strain NS-810 by submerged fermentation.
参考文献/References:
[1]申鸿,张龙,王兵,等.蚕体和蛹粉代料培养基上的蛹虫草生长状况与品质检测[J].蚕业科学,2012,38(1):130-134.
[2]袁明生,孙佩琼.中国蕈菌原色图集[M].成都:四川科学技术出版社,2007.
[3]温鲁,尹起范,唐玉玲,等.蚕虫草与有关虫草活性成分检测比较[J].食品科学,2004,25(8):155-157.
[4]NI H,ZHOU X H,LI H H,et a1.Column chromatographic extraction and preparation of cordycepin from Cordyceps militaris waster medium[J].Journal of Chromatography B Analytical Technologies in the Biomedical & Life Sciences,2009,877(22):2135-2141.
[5]岳翠翠,沈健增,蔡宇杰,等.蛹虫草Cordyceps militaris JN168产虫草素液态发酵条件的优化[J].食品与生物技术学报,2013,32(2):135-141.
[6]文庭池,李光荣,康冀川,等.蛹虫草液体种制备及发酵生产菌丝体和虫草菌素工艺优化[J].食品科学,2012,33(5):144-149.
[7]刘桂君,周思静,杨素玲,等.蛹虫草中虫草素的研究进展[J].食品科学,2013,34(21):408-412.
[8]王兰青,高玉千,戚元成,等.菇液体菌种摇瓶和发酵罐生长动力学模型的比较[J].食品与生物技术学报,2010,29(6):929-933.
[9]温鲁,夏敏,宋虎卫,等.液体培养蛹虫草虫草素和腺苷的代谢量[J].微生物学通报,2005,32(3):91-94.
[10]陈磊.蛹虫草(Cordyceps militaris)生物学特性及发酵研究[D].北京:北京协和医学院,2009.
[11]XIAO J H,CHEN D X，XIAO Y,et a1.Optimization of submerged culture conditions for mycelial polysaccharide production in Cordyceps pruinosa[J].Process Biochemistry,2004,39:2241-2247.
[12]叶晶晶,张剑飞,曹宁宁,等.添加蚕蛹粉和改变营养液pH对人工培养蛹虫草子实体产量及虫草素含量的影响[J].蚕业科学,2013,39(3):624-627.
[13]雷坤,柯轶,毛宁.蛹虫草Fjnu-01高产虫草素的液体培养基优化[J].药物生物技术,2011,18(6):504-508.
[14]欧阳召,王春梅,陶志.适合虫草素积累的蛹虫草液体培养条件的优化[J].中国食用菌,2012,31(1):26-28.
相似文献/References:
[1]孙露露,李焕端,张至远,等.蛹虫草甜米酒的生产工艺研究[J].北方园艺,2014,38(05):125.
SUN Lu-lu,LI Huan-duan,ZHANG Zhi-yuan,et al.Study on Production Techniques of Cordyceps Militaris Sweet Rice Wine[J].Northern Horticulture,2014,38(05):125.
[2]方华舟.蛹虫草菌种培养性状与出草结果关系的研究[J].北方园艺,2013,37(12):161.
FANG Hua-zhou.Study on the Relationship Between Cultural Characteristics and Results of Weeds of Cordyceps militaris[J].Northern Horticulture,2013,37(05):161.
[3]殷东林,段鸿斌,王锐丽.蛹虫草有性生殖研究[J].北方园艺,2014,38(03):130.
YIN Dong-lin,DUAN Hong-bin,WANG Rui-li.Study on the Sexual Reproduction of Cordyceps militaris[J].Northern Horticulture,2014,38(05):130.
[4]刘 洋,孙来玉,陆云华,等.九株蛹虫草菌株的生物学性状研究[J].北方园艺,2013,37(20):140.
LIU Yang,SUN Lai-yu,LU Yun-hua,et al.Study on the Biological Characteristics of Nine Cordyceps militaris Strains[J].Northern Horticulture,2013,37(05):140.
[5]付彦凯,冷云伟,曹文化.蛹虫草的人工培育历程[J].北方园艺,2012,36(04):178.
FU Yan-kai,LENG Yun-wei,CAO Wen-hua.Artificial Cultivation Progress of Cordyceps militaris[J].Northern Horticulture,2012,36(05):178.
[6]暴增海,贾超艳,江倩倩,等.真姬菇、鸡汁菇和蛹虫草培养特性的观察与生长量的测定[J].北方园艺,2012,36(23):162.
BAO Zeng-hai,JIA Chao-yan,JIANG Qian-qian,et al.Cultivation Characteristics and Growth of Hypsizygus marmoreus，Jizhigu and Cordyceps militaris[J].Northern Horticulture,2012,36(05):162.
[7]郑素月,郝俊灵,王建伟.继代培养对蛹虫草菌丝生长速度及酯酶同工酶的影响[J].北方园艺,2012,36(10):136.
ZHENG Su-yue,HAO Jun-ling,WANG Jian-wei.The Influences of Subculture of Cordceps militaris to Mycelial Growth Rate and Esterase Isozyme[J].Northern Horticulture,2012,36(05):136.
[8]方华舟.不同蛹虫草优质母种分离方式与菌种性能比较[J].北方园艺,2014,38(15):142.
FANG Hua-zhou.Comparison of Separation Method and Performance Comparison Among Different High Quality Stock Culture of Cordyceps militaris[J].Northern Horticulture,2014,38(05):142.
[9]林群英,吴亮亮,张锋伦,等.蛹虫草栽培过程中胞外蛋白酶活性变化研究[J].北方园艺,2014,38(17):148.
LIN Qun-ying,WU Liang-liang,ZHANG Feng-lu,et al.Study on Extracellular Protease Activity Change During Fruitbody Growth of Cordyceps militaris[J].Northern Horticulture,2014,38(05):148.
[10]张筱梅,朱维红,贾春凤.二株茶薪菇菌丝多糖深层发酵的培养特征比较与条件控制[J].北方园艺,2014,38(23):127.
ZHANG Xiao-mei,ZHU Wei-hong,JIA Chun feng.Culture Characteristic Comparison and Condition Control of Fermentation of Two Polysaccharides Agrocybe chaxingu[J].Northern Horticulture,2014,38(05):127.
[11]林群英,封义,姜艳,等.蛹虫草液体培养产纤溶酶条件优化研究[J].北方园艺,2015,39(24):143.[doi:10.11937/bfyy.201524039]
LIN Qunying,FENG Yi,JIANG Yan,et al.Optimization of Submerged Culture Conditions for Production of Fibrinolytic Enzyme by Cordyceps militaris[J].Northern Horticulture,2015,39(05):143.[doi:10.11937/bfyy.201524039]
备注/Memo
第一作者简介：张楠（1980-），男，博士，副教授，研究方向为微生物学与食用菌种植。E-mail:nan25000@163.com.责任作者：段辉国(1967-)，男，硕士，教授，硕士生导师，研究方向为生物化学。E-mail:duanhuiguo6@163.com.基金项目：四川省教育厅重点资助项目（11ZA029）；川西珍稀植物资源研究与利用创新团队资助项目（14TD0025）。