黑皮鸡枞液体菌种发酵罐培养条件的优化
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摘要
【目的】优化黑皮鸡枞液体菌种发酵罐培养条件,为大规模培养黑皮鸡枞液体栽培菌种及提高黑皮鸡枞菌产量和品质提供参考依据。【方法】采用液体发酵罐培养技术,以黑皮鸡枞液体菌种的菌球直径、均匀度、菌丝干重、漆酶、纤维素酶和多酚氧化酶活性为指标,通过单因素试验和均匀试验,优化碳(C)源、氮(N)源、碳氮添加量比(C/N)、接种量、装液量、培养温度和培养时间,筛选出黑皮鸡枞液体菌种发酵罐培养的最佳培养基和发酵工艺,并开展液体菌种栽培试验。【结果】黑皮鸡枞液体菌种发酵罐培养的最佳培养基为20.0 g葡萄糖+4.0 g高粱粉+3.0 g K_2HPO_4+1.0 g MgSO_4+2片维生素B_1+1000.0 mL蒸馏水,pH 6.5;发酵工艺:装液量12 L,接种量12%,培养温度25℃,转速90 r/min,培养时间100 h,罐压0.3 MPa,通气量0.9 m3/h。栽培试验结果表明,在最佳培养基和培养工艺条件下获得的液体菌种其栽培黑皮鸡枞菌丝生长速度快,满袋时间为29 d,平均每袋产量约360.0 g,生物学转化率达78.0%。【结论】将液体菌种发酵罐培养技术与工厂化栽培技术有机结合,可实现黑皮鸡枞菌的规模化、标准化、现代化和工厂化生产。
【Objective】The culture conditions of fermenting tanks for Oudemansiella raphanipes liquid strain were optimized,which provided reference for large-scale cultivation of liquid culture strains and the improvement of the yield and quality of O. raphanipes. 【Method】Using liquid fermenter culture technology,the ball diameter,uniformity,dry weight of the mycelium,laccase activity,cellulase activity and polyphenol oxidase activity of the liquid strain of O.raphanipes were used as indicators. Through single factor and uniform experiments,optimized carbon(C)source,nitrogen(N)source,carbon and nitrogen add ratio(C/N),inoculum amount,volume of liquid,culture temperature and incubation time,the optimum medium and fermentation process of O. raphanipes liquid strains were screened,and the cultivation experiment of liquid strain was carried out.【Result】The best medium for fermentation of O. raphanipes liquid strains by fermenter:glucose 20.0 g+sorghum powder 4.0 g+K_2 HPO_4 3.0 g+MgSO_4 1.0 g+vitamin B_1 2 tablets+distilled water 1000.0 mL,pH 6.5. Fermentation process:liquid content 12 L,inoculation amount 12%,culture temperature 25 ℃,stirring speed90 r/min,culture time 100 h,tank pressure 0.3 MPa,ventilation volume 0.9 m3/h. The results of the cultivation experiment showed that the mycelium of the O. raphanipes cultivated in the liquid culture under the optimal medium and the optimum process conditions had a fast growth rate,the mycelium full bag time was 29 d,and the average yield each bag was about360.0 g. The biological conversion rate reached 78.0%.【Conclusion】It is possible to realize the large-scale,standardized and modern industrial production of O. raphanipes by combining the liquid strain fermenter production technology with the industrial cultivation technology.
【Objective】The culture conditions of fermenting tanks for Oudemansiella raphanipes liquid strain were optimized,which provided reference for large-scale cultivation of liquid culture strains and the improvement of the yield and quality of O. raphanipes. 【Method】Using liquid fermenter culture technology,the ball diameter,uniformity,dry weight of the mycelium,laccase activity,cellulase activity and polyphenol oxidase activity of the liquid strain of O.raphanipes were used as indicators. Through single factor and uniform experiments,optimized carbon(C)source,nitrogen(N)source,carbon and nitrogen add ratio(C/N),inoculum amount,volume of liquid,culture temperature and incubation time,the optimum medium and fermentation process of O. raphanipes liquid strains were screened,and the cultivation experiment of liquid strain was carried out.【Result】The best medium for fermentation of O. raphanipes liquid strains by fermenter:glucose 20.0 g+sorghum powder 4.0 g+K_2 HPO_4 3.0 g+MgSO_4 1.0 g+vitamin B_1 2 tablets+distilled water 1000.0 mL,pH 6.5. Fermentation process:liquid content 12 L,inoculation amount 12%,culture temperature 25 ℃,stirring speed90 r/min,culture time 100 h,tank pressure 0.3 MPa,ventilation volume 0.9 m3/h. The results of the cultivation experiment showed that the mycelium of the O. raphanipes cultivated in the liquid culture under the optimal medium and the optimum process conditions had a fast growth rate,the mycelium full bag time was 29 d,and the average yield each bag was about360.0 g. The biological conversion rate reached 78.0%.【Conclusion】It is possible to realize the large-scale,standardized and modern industrial production of O. raphanipes by combining the liquid strain fermenter production technology with the industrial cultivation technology.
引文
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徐兵,张雅君,冀宏.2017.黑皮鸡枞菌丝体液体发酵条件优化[J].常熟理工学院学报,31(2):104-108.[Xu B,Zhang Y J,Ji H.2017.Optimization of liquid fermentation conditions for the mycelia of Termitomyces Heim[J].Journal of Changshu Institute Technology,31(2):104-108.]
薛林浩,王晓东,蔡虹,范红春,党会,黎勇.2013.黑皮鸡枞菌液体培养基优化[J].内江师范学院学报,28(10):24-27.[Xue L H,Wang X D,Cai H,Fan H C,Dang H,Li Y.2013.Optimization of liquid culture medium for Termitomyces Heim[J].Journal of Neijiang Normal University,28(10):24-27.]
曾先富,陈阳婷,熊维全,罗小波.2012.鸡枞菌菌丝体培养试验[J].食用菌,34(6):9-10.[Zeng X F,Chen Y T,Xiong W Q,Luo X B.2012.Experiment on mycelium culture of Termitomyces[J].Edible Fungi,34(6):9-10.]
朱海潇.2008.食用菌产漆酶能力的比较及漆酶性能的研究[D].福州:福建农林大学.[Zhu H X.2008.The comparison for the capacity laccase production in various types of edible fungi and the characterization of laccase[D].Fuzhou:Fujian Agriculture and Forestry University.]
Saha T.2008.Cellobiose dehydrogenase production by the mycelial culture of the mushroom Termitomyces clypeatus[J].Process Biochem,43(6):634-641.
Kirk P M,Cannon P F,David J C,Stalpers J A.2001.Ainsworth&Bisby’s Dictionary of the Fungi[M].Wallingford:CAB1 Publishing.
戴建清,曾志恒.2012.食用菌液体菌种研究现状及发展趋势[J].中国食用菌,31(5):1-3.[Dai J Q,Zeng Z H.2012.Research status and development trends of liquid spawn of edible mushroom[J].Edible Fungi of China,31(5):1-3.]
付子艳,李荣春.2009.云南鸡枞菌属关系的初步研究[J].福建农林大学学报(自然科学版),38(3):271-274.[Fu ZY,Li R C.2009.Initial research in phylogenetics relationship of Termitomyces Heim in Yunnan,China[J].Journal of Fujian Agriculture and Forestry University(Natural Science Edition),38(3):271-274.]
郭尚,王慧娟.2013.食用菌深层发酵技术及其应用[J].山西农业科学,41(8):885-888.[Guo S,Wang H J.2013.Submerged fermentation technology of edible fungus and its application[J].Journal of Shanxi Agricultural Sciences,41(8):885-888.]
胡尚勤,刘天贵,李贤柏.2008.鸡枞菌的培养条件研究[J].食品与生物技术学报,27(1):67-70.[Hu S Q,Liu T G,Li X B.2008.Research of the nutrition requirement of the Termitomyces albuminosu[J].Journal of Food Science and Biotechnology,27(1):67-70.]
胡忠策,郑晓冬.2002.鸡枞菌液体深层发酵的研究[J].菌物系统,21(1):98-101.[Hu Z C,Zheng X D.2002.Study on submerged fermentation of abieties[J].Mycosystema,21(1):98-101.]
李艳丽.2018.鸡枞菌培养基优化及人工栽培技术研究[D].汉中:陕西理工大学.[Li Y L.2018.Research on optimization and artificial cultivation technology of Termitomyces albuminosus culture[D].Hanzhong:Shaanxi University of Technology.]
罗晓妙.2010.液态发酵产鸡枞菌菌丝体的条件优化[J].食品科技,35(3):29-32.[Luo X M.2010.Optimization of liquid-state fermentation conditions for producing mycelium of Termitomyces albuminousus[J].Food Science and Technology,35(3):29-32.]
聂晓东.2009.鸡枞菌丝体及其皂苷的深层发酵条件的优化[D].无锡:江南大学.[Nie X D.2009.Optimization of submerged culture in mycelia and saponin in Termitomyces albuminosus[D].Wuxi:Jiangnan University.]
孙月娥,张俊韬,王卫东.2011.金针菇中多酚氧化酶活性测定及其护色研究[J].食品工业,(11):68-70.[Sun Y E,Zhang J T,Wang W D.2011.PPO activity determination and antibrowning measurement for Flammulina velutipes[J].Food Industry,(11):68-70.]
苏跃稳,路鹏,郭群群,王健.2016.食用菌液体深层发酵的研究进展[J].食品安全质量检测学报,7(2):645-650.[Su Y W,Lu P,Guo Q Q,Wang J.2016.Progress in edible fungus by submerged fermentation[J].Journal of Food Safety and Quality,7(2):645-650.]
王谦,胡卫静.2016.大型食药用真菌深层发酵研究进展[J].食品安全质量检测学报,7(3):1240-1246.[Wang Q,Hu W J.2016.Research progress on submerged fermentation of macroscopic edible fungi[J].Journal of Food Safety and Quality,7(3):1240-1246.]
熊亚,李敏杰.2013.鸡枞菌菌丝体基础培养基的优化[J].食品与机械,29(4):185-189.[Xiong Y,Li M J.2013.Optimization of basic medium for mycelium of Collybia albuminosa by response surface methodology[J].Food and Machinery,29(4):185-189.]
徐兵,张雅君,冀宏.2017.黑皮鸡枞菌丝体液体发酵条件优化[J].常熟理工学院学报,31(2):104-108.[Xu B,Zhang Y J,Ji H.2017.Optimization of liquid fermentation conditions for the mycelia of Termitomyces Heim[J].Journal of Changshu Institute Technology,31(2):104-108.]
薛林浩,王晓东,蔡虹,范红春,党会,黎勇.2013.黑皮鸡枞菌液体培养基优化[J].内江师范学院学报,28(10):24-27.[Xue L H,Wang X D,Cai H,Fan H C,Dang H,Li Y.2013.Optimization of liquid culture medium for Termitomyces Heim[J].Journal of Neijiang Normal University,28(10):24-27.]
曾先富,陈阳婷,熊维全,罗小波.2012.鸡枞菌菌丝体培养试验[J].食用菌,34(6):9-10.[Zeng X F,Chen Y T,Xiong W Q,Luo X B.2012.Experiment on mycelium culture of Termitomyces[J].Edible Fungi,34(6):9-10.]
朱海潇.2008.食用菌产漆酶能力的比较及漆酶性能的研究[D].福州:福建农林大学.[Zhu H X.2008.The comparison for the capacity laccase production in various types of edible fungi and the characterization of laccase[D].Fuzhou:Fujian Agriculture and Forestry University.]
Saha T.2008.Cellobiose dehydrogenase production by the mycelial culture of the mushroom Termitomyces clypeatus[J].Process Biochem,43(6):634-641.
Kirk P M,Cannon P F,David J C,Stalpers J A.2001.Ainsworth&Bisby’s Dictionary of the Fungi[M].Wallingford:CAB1 Publishing.