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[1]郭梦圆,高艳华,郑雨璋,等.冬虫夏草内生菌NS2-B1发酵培养基筛选[J].应用与环境生物学报,2017,23(05):939-944.[doi:10.3724/SP.J.1145.2016.11013]
 GUO Mengyuan,GAO Yanhua,ZHENG Yuzhang,et al.Screening of liquid fermentation media of an endophytic fungus NS2-B1 isolated from Ophiocordyceps sinensis[J].Chinese Journal of Applied & Environmental Biology,2017,23(05):939-944.[doi:10.3724/SP.J.1145.2016.11013]
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冬虫夏草内生菌NS2-B1发酵培养基筛选()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
23卷
期数:
2017年05期
页码:
939-944
栏目:
生物活性化合物的发现与开发专栏
出版日期:
2017-10-25

文章信息/Info

Title:
Screening of liquid fermentation media of an endophytic fungus NS2-B1 isolated from Ophiocordyceps sinensis
作者:
郭梦圆高艳华郑雨璋王萍夏飞周选围
1上海交通大学农业与生物学院植物生物技术研究中心,农业部都市农业(南方)重点实验室 上海 200240 2陕西科技大学食品与生物工程学院 西安 710021 3天津理工大学化学化工学院 天津 300384
Author(s):
GUO Mengyuan1 GAO Yanhua1 ZHENG Yuzhang1 WANG Ping3 XIA Fei2** & ZHOU Xuanwei1**
1Ministry of Agriculture Key Laboratory of Urban Agriculture (South), Plant Biotechnology Research Center, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China 2 School of Food and Biological Engineering, Shaanxi University of Science & Technology, Xi’An 710021, China 3 College of Chemistry and Chemical Engineering, Tianjin University of Technology, Tianjin, 300384, China
关键词:
冬虫夏草内生真菌液体培养基正交试验代谢产物
Keywords:
Ophiocordyceps sinensis endophytic fungus liquid media orthogonal test metabolites
分类号:
Q93-3:Q929
DOI:
10.3724/SP.J.1145.2016.11013
摘要:
以核苷类物质为目标产物筛选冬虫夏草内生真菌NS2-B1生产具有和冬虫夏草相似活性成分的培养基. 通过单因素实验,分别从5种碳源(蔗糖、果糖、葡萄糖、麦芽糖、淀粉)和5种氮源(尿素、硝酸铵、牛肉膏、酵母膏、蛋白胨)中各筛选出2种最佳碳源和氮源;然后通过4因素3水平正交实验L9(34),以代谢产物总核苷(尿苷、鸟苷、腺苷和虫草素)为指标筛选最佳的碳、氮源组合. 结果显示:NS2-B1液体发酵培养基组合为麦芽糖2%、淀粉1.5%、酵母膏1.5%、蛋白胨1%,附加KH2PO4 0.8 g/L和MgSO4.7H2O 0.8 g/L;利用该培养基在25 ℃、210 r/min条件下震荡培养,5 d后菌丝体发酵液中总核苷的产量达到(92.62 ± 4.19)μg/mL. 综上,从天然冬虫夏草中分离的内生真菌NS2-B1具有替代冬虫夏草生产核苷类物质的潜力. (图4 表4 参30)
Abstract:
Based on nucleosides as the target produces, the suitable liquid media for fermenting an endophytic fungus NS2-B1 isolated from Ophiocordyceps sinensis was investigated to produce active ingredients which were similar to O. sinensis. Two optimum carbon sources and two optimum nitrogen sources were selected from five carbon sources (including sucrose, fructose, glucose, maltose and starch) and five nitrogen sources (including urea, beef extract, yeast extractand peptone) by one-factor-at-a-time experiments. The suitable culture medium for producing the nucleosides was studied by orthogonal test. Table L9(34) was used to test four factors with three levels. The nucleosides (uridine, guanosine, adenosine, cordycepin) of O. sinensis metabolites in the mycelial fermentation broth were determined by HPLC. The results showed that the suitable liquid medium for the endophytic fungus isolated from O. sinensis consisted of 2% maltose, 1.5% starch, 1.5% yeast extract, 1% peptone, plus to 0.8 g/L KH2PO4 and 0.8 g/L MgSO4.7H2O. The yield of the nucleosides in the mycelial fermentation broth reached to 92.62 ± 4.19 μg/mL under the conditions of 25 ℃ and 210 r/min for 5 days. Therefore, the endophytic fungus NS2-B1 had the potential to replace the Ophiocordyceps sinensis in producing nucleosides.

参考文献/References:

1 董彩虹, 李文佳, 李增智, 闫文娟, 李泰辉, 刘杏忠. 我国虫草产业发展现状、问题及展望——虫草产业发展金湖宣言[J]. 菌物学报, 2016, 35 (1): 1-15 [Dong CH, Li WJ, Li ZZ, Yan WJ, Li TH. Cordyceps industry in China: current status, challenges and perspectives - Jinhu declaration for cordyceps industry development [J]. Mycosystema, 2016, 35 (1): 1-15]
2 Zhou XW, Li LJ, Tian EW. Advances in research of the artificial cultivation of Ophiocordyceps sinensis in China [J]. Crit Rev Biotechnol, 2014, 34 (3): 233-243
3 Lo HC, Hsieh C, Lin FY, Hsu TH. A systematic review of the mysterious caterpillar fungus Ophiocordyceps sinensis in Dong-ChongXiaCao (冬蟲夏草 Dōng Chóng Xià Cǎo) and related bioactive ingredients [J]. J Tradit Complem Med, 2013, 3 (1): 16-32
4 Paterson RRM. Cordyceps-A traditional Chinese medicine and another fungal therapeutic biofactory? [J]. Phytochemistry, 2008, 69 (7): 1469-1495
5 Zhou XW, Gong Z, Su Y, Lin J, Tang KX. Cordyceps fungi: natural products, pharmacological functions and developmental products [J]. J Pharm Pharmacol, 2009, 61 (3): 279-291
6 Weckerle CS, Yang Y, Huber FK, Li Q. People, money, and protected areas: the collection of the caterpillar mushroom Ophiocordyceps sinensis in the Baima Xueshan Nature Reserve, Southwest China [J]. Biodivers Conserv, 2010, 19 (9): 2685-2698
7 Zhang Y, Xu L, Zhang S, Liu X, An Z, Wang M, Guo Y. Genetic diversity of Ophiocordyceps sinensis, a medicinal fungus endemic to the Tibetan Plateau: implications for its evolution and conservation [J]. BMC Evol Biol, 2009, 9 (1): 290
8 李文才, 邱建军, 邱锋. 西藏那曲地区冬虫夏草资源可持续开发利用研究[J]. 生态经济, 2011 (6): 112-114 [Li WC, Qiu JJ, Qiu F. Research on sustainable utilization of Cordyceps sinensis resources in Naqu Region of Xizang Zizhiqu [J]. Ecol Econ, 2011 (6): 112-114]
9 Xia F, Liu Y, Guo MY, Shen GR, Lin J, Zhou XW. Pyrosequencing analysis revealed complex endogenetic microorganism community from natural DongChong XiaCao and its microhabitat [J]. BMC Microbiol, 2016, 16 (1): 196
10 Xia F, Chen X, Guo MY, Bai XH, Liu Y, Shen GR, Li YL, Lin J, Zhou XW. High-throughput sequencing-based analysis of endogenetic fungal communities inhabiting the Chinese Cordyceps reveals unexpectedly high fungal diversity [J]. Sci Rep, 2016, 6: 33437
11 Xia F, Liu Y, Shen GR, Guo LX, Zhou XW. Investigation and analysis of microbiological communities in natural Ophiocordyceps sinensis [J]. Can J Microbiol, 2014, 61 (2): 104-111
12 Stierle AG. Taxol and taxane production by Taxomyces andreanae, an endophytic fungus of Pacific yew [J]. Science, 1993, 260: 214-216
13 Zhou XW, Zhu H, Liu L, Lin J, Tang KX. A review: recent advances and future prospects of taxol-producing endophytic fungi [J]. App Microbiol Biotechnol, 2010, 86 (6): 1707-1717
14 金成, 邬晓敏, 陈高阳, 金水宝的临床应用[J]. 首都医药, 2006, 12 (18): 42-43 [Jin C, Wu XM, Chen GY. Clinical application of Jinshuibao [J]. Capital Med, 2006, 12 (18): 42-42]
15 Roger PA, Heong KL, Teng PS, Hawksworth DL. The biodiversity of microorganisms and invertebrates: its role in sustainable agriculture [C]. Proceedings of the 1st Workshop on the Ecological Foundations of Sustainable Agriculture (WEFSA’91), 1991: 117-136
16 梁宗琦. 真菌次生代谢产物多样性及其潜在应用价值[J]. 生物多样性, 1999, 7 (2): 145-150 [Liang ZQ, The diversity of fungal secondary metabolites and their potential applications [J]. Chin Biodiv, 1999, 7 (2): 145-150]
17 张永杰, 孙炳达, 张姝, 旺姆, 刘杏忠, 巩文峰. 分离自冬虫夏草可培养真菌的多样性研究[J]. 菌物学报, 2010, 29 (4): 518-527 [Zhang YJ, Sum BD, Zhang S, Wang M, Liu XZ, Gong WF. Mycobiotal investigation of natural Ophiocordyceps sinensis based on culture-dependent investigation [J]. Mycosystema, 2010, 29 (4): 518-527]
18 姚艺桑, 朱佳石. 中药冬虫夏草和所含多种冬虫夏草菌拉丁名混用的历史和现状[J]. 中国中药杂志, 2016, 41 (7): 1361-1366 [Yao YS, Zhu JS. Indiscriminate use of Latin name for natural Cordyceps sinensis insect-fungi complex and multiple Ophiocordyceps sinensis fungi [J]. China J Chin Mater Med, 2016, 41 (7): 1361-1366]
19 张晓芳, 陈琼, 刘慧娜, 彭璐璐, 朱艳丽. 探析冬虫夏草无性型发酵技术研究新进展[J]. 生物技术世界, 2015 (1): 57 [Zhang XF, Chen Q, Liu HN, Peng LL, Zhu YL. New advances in the fermentation technique of Cordyceps sinensis anamorph [J]. Biotech World, 2015 (1): 57]
20 车永胜. 特殊生境真菌来源的新结构活性物质研究[J]. 国际药学研究杂志, 2011, 38 (1): 12-27 [Che YS. Discovery of new bioactive natural products from fungi of unique ecological niches [J]. J Int Pharm Res, 2011, 38 (1): 12-27]
21 胡丰林, 李增智. 虫草及相关真菌的次生代谢产物及其活性[J]. 菌物学报, 2007, 26 (4): 607-632 [Hu FL, Li ZZ. Secondary metabolites and their bioactivities of cordyceps and its related fungi [J]. Mycosystema, 2007, 26 (4): 607-632]
22 Guo H, Hu H, Liu S, Liu X, Zhou Y, Che Y. Bioactive p-Terphenyl derivatives from a Cordyceps-colonizing isolate of Gliocladium sp. [J]. J Nat Prod, 2007, 70 (9): 1519-1521
23 Yamada T, Iwamoto C, Yamagaki N, Yamanouchi T, Minoura K, Yamori T, Uehara Y, Andoh T, Umemura K, Numata A. Leptosins M-N1, cytotoxic metabolites from a Leptosphaeria species separated from a marine alga. Structure determination and biological activities [J]. Tetrahedron, 2002, 58 (3): 479-487
24 Zhang Y, Liu S, Che Y, Liu X. Epicoccins A–D, Epipolythiodioxo-piperazines from a Cordyceps-colonizing isolate of Epicoccum nigrum [J]. J Nat Prod, 2007, 70 (9): 1522-1525
25 魏江春, 魏鑫丽, 郑维发, 郭威, 柳润东, 现代工业化培植的冬虫夏草物种鉴定与成分检测[J]. 菌物学报, 2016, 35 (4): 404-410 [Wei JC, Wei XL, Zheng WF, Guo W, Liu RD, Species identification and component detection of Ophiocordyceps sinensis cultivated by modern industry [J]. Mycosystema, 2016, 35 (4): 404-410]
26 Liu Y, Yuan WN, Zhou XW, Optimization of submerged culture conditions for the mycelial biomass and bioactive metabolites production by Cordyceps militaris [J]. J Pure Appl Microbiol, 2014, 8 (6): 4245-4256
27 Huang L, Li QZ, Chen Y, Wang X, Zhou XW, Determination and analysis of cordycepin and adenosine in the products of Cordyceps spp. [J]. Afr J Microbiol Res, 2009, 3 (12): 957-961
28 Xia F, Liu Y, Guo MY, Shen GR, Lin J, Zhou XW. Pyrosequencing analysis revealed complex endogenetic microorganism community from natural DongChong XiaCao and its microhabitat [J]. BMC Microbiol, 2016, 6 (1): 196
29 Xia F, Chen X, Guo MY, Bai XH, Liu Y, Shen GR, Li YL, Lin J, Zhou XW. High-throughput sequencing-based analysis of endogenetic fungal communities inhabiting the Chinese Cordyceps reveals unexpectedly high fungal diversity [J]. Sci Rep, 2016, 6: 33437
30 Xia F, Liu Y, Shen GR, Lin J, Zhou XW. Investigation and analysis of microbiological communities in natural Ophiocordyceps sinensis [J]. Can J Microbiol, 2015, 61 (2): 104-111

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更新日期/Last Update: 2017-10-25