六妹羊肚菌工厂化制备液体菌种繁育条件的优化
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摘要
为适合大规模工厂化制备羊肚菌液体菌种,以提高菌丝体生物量为目的,控制菌丝球直径,使用六妹羊肚菌Morchellasextelata,采用单因素、Plackett-Burman、响应面Box-Behnken实验摇瓶培养优化液体菌种制备条件,发酵罐扩大培养,具体参数为玉米淀粉2.5%、麸皮6%、酵母粉0.4%、葡萄糖2.5%、蛋白胨0.25%、磷酸二氢钾0.1%、硫酸镁0.05%、接种量10%、pH 5.5、转速200–500r/min、温度25℃、培养1–3d,在该条件下羊肚菌菌丝体干重可达16mg/mL,菌丝球直径降至1.5mm以下,密度达到2967个/mL,从母种到栽培种仅需13–19d。本研究结果为羊肚菌工厂化液体菌种制备和栽培提供了理论基础。
Single factor experiment,Plackett-Burman experiment and response surface Box-Behnkeh experiment were implemented for optimization of massive industrialized production of smaller-pelletized Morchella sextelata liquid spawn.The optimized medium contained 2.5%corn starch,6%bran,0.4%yeast powder,2.5%glucose,0.25%peptone,0.1%KH_2PO_4,and 0.05%MgSO_4.Under the conditions of inoculation volume of 10%,pH value of 5.5,rotation speed of 200–500r/min,and incubation temperature of 25°C,the biomass of Morchella sextelata reached 16mg/mL in 3 days.The diameter of mycelium pellet reduced to1.5mm and the density reached 2 967/mL.The culture period from the stock culture to the applicable spawn was 13–19 days.Our research offered reference for Morchella sextelata liquid spawn industrialized production.
Single factor experiment,Plackett-Burman experiment and response surface Box-Behnkeh experiment were implemented for optimization of massive industrialized production of smaller-pelletized Morchella sextelata liquid spawn.The optimized medium contained 2.5%corn starch,6%bran,0.4%yeast powder,2.5%glucose,0.25%peptone,0.1%KH_2PO_4,and 0.05%MgSO_4.Under the conditions of inoculation volume of 10%,pH value of 5.5,rotation speed of 200–500r/min,and incubation temperature of 25°C,the biomass of Morchella sextelata reached 16mg/mL in 3 days.The diameter of mycelium pellet reduced to1.5mm and the density reached 2 967/mL.The culture period from the stock culture to the applicable spawn was 13–19 days.Our research offered reference for Morchella sextelata liquid spawn industrialized production.
引文
Abdullah N,Rozila I,Norjuliza MKJ,Annuar MMS,2013.Production of liquid spawn of an edible grey oyster mushroom,Pleurotus pulmonarius(Fr.)Quél.by submerged fermentation and sporophore yield on rubber wood sawdust.Scientia Horticulturae,161:65-69
Dai JQ,Zeng ZH,2012.Research status and development trend of liquid strains of edible fungi.Chinese Mushroom,2012(5):1-3(in Chinese)
Deacon J,2006.Fungal biology.Blackwell Publishing Ltd.,Oxford.1-366
Dong CH,Yao YJ,2010.Nutritional requirements of mycelial growth of Cordyceps sinensis in submerged culture.Journal of Applied Microbiology,99(3):483-492
Dubois M,Gilles K,Hamilton JK,Rebers PA,Smith F,1951.Colorimetric method for the determination of sugars.Nature,168(4265):167
Fang QH,Zhong JJ,2002.Effect of initial pH on production of ganoderic acid and polysaccharide by submerged fermentation of Ganoderma lucidum.Process Biochemistry,37(7):769-774
Francis F,Abdulhameed S,Madhavan N,Sumitra R,Ashok P,Sanjoy G,George S,2003.Use of response surface methodology for optimizing process parameters for the production of alpha-amylase by Aspergillus oryzae.Biochemical Engineering Journal,15(2):107-115
Fu YQ,Yin LF,Zhu HY,Jiang R,Li S,Xu Q,2014.Effects of pellet characteristics on L-lactic acid fermentation by R.oryzae:pellet morphology,diameter,density,and interior structure.Applied Biochemistry and Biotechnology,174(6):2019-2030
Ji H,Zhao LM,2009.Methods and ideas for improving industrial production technology of edible fungi.Chinese Mushroom,28(6):56-58(in Chinese)
Jin ZX,Wang YL,Wang PZ,Sun YB,Zhang ZS,2014.Optimization of culture condition for liquid fermentation of morchella.Journal of Dalian University of Technology,2014(6):416-419(in Chinese)
Kirsch LDS,Ana JPDM,Maria FST,2016.Production of mycelial biomass by the Amazonian edible mushroom Pleurotus albidus.Brazilian Journal of Microbiology,47(3):658-664
Li H,Zhang M,Li C,Zhang JJ,2018.Study on liquid culture technology of morels.Journal of Shenyang Agricultural University,2018(2):227-233(in Chinese)
Li JS,Ma CQ,Ma YH,Li Y,Zhou W,Xu P,2007.Medium optimization by combination of response surface methodology and desirability function:an application in glutamine production.Applied Microbiology and Biotechnology,74(3):563-571
Liu QY,Qi J,Zhou HY,Sun B,Wu HL,2018.Brief analysis on current situation and development of industrial production and application of edible fungus liquid strain.Journal of Dalian University of Technology,40(6):8-10(in Chinese)
Saad N,Abdeshahian P,Kalil MS,Yusoff WM,Hamid AA,2014.Optimization of aeration and agitation rate for lipid and gamma linolenic acid production by Cunninghamella bainieri 2A1 in submerged fermentation using response surface methodology.Scientific World Journal,2014(1):280146
Srinivas MRS,Nagin C,Lonsane BK,1994.Use of Plackett-Burman design for rapid screening of several nitrogen sources,growth/product promoters,minerals and enzyme inducers for the production of alpha-galactosidase by Aspergillus niger MRSS 234 in solid state fermentation system.Bioprocess Engineering,10(3):139-144
Vishvakarma R,Abha M,2017.Production of a protease inhibitor from edible mushroom Agaricus bisporus and its statistical optimization by response surface methodology.Preparative Biochemistry&Biotechnology,47(5):450-457
Wang D,Akiyoshi S,Motoyuki S,2001.Biological efficiency and nutritional value of Pleurotus ostreatus cultivated on spent beer grain.Bioresource Technology,78(3):293-300
Wu JZ,Peter CKC,Ka HW,Nian LH,2004.Studies on submerged fermentation of Pleurotus tuber-regium(Fr.)Singer.Part 2:effect of carbon-to-nitrogen ratio of the culture medium on the content and composition of the mycelial dietary fibre.Food Chemistry,85(1):101-105
Xiao JH,Dai XC,Wei HW,Xi JH,Ying Q,Zong QL,2006.Enhanced simultaneous production of mycelia and intracellular polysaccharide in submerged cultivation of Cordyceps jiangxiensis using desirability functions.Process Biochemistry,41(8):1887-1893
Zhang RY,Dan DH,Xiao TM,Shi GL,Jin GG,Qing XH,2014.Adopting stick spawn reduced the spawn running time and improved mushroom yield and biological efficiency of Pleurotus eryngii.Scientia Horticulturae,175(1):156-159
Zhu LW,Hong ML,Dong SL,Ya JT,2007.Submerged culture of mushrooms in bioreactors-challenges,current state-of-the-art,and future prospects.Food Technology&Biotechnology,45(3):221-229
戴建清,曾志恒,2012.食用菌液体菌种研究现状及发展趋势.中国食用菌,2012(5):1-3
李红,张敏,李超,张季军,2018.羊肚菌液体培养工艺研究.沈阳农业大学学报,2018(2):227-233
金朝霞,王云龙,王培忠,孙业兵,张宗申,2014.羊肚菌液体发酵培养条件优化.大连工业大学学报,2014(6):416-419
刘启燕,戚俊,周洪英,孙波,吴洪丽,2018.食用菌液体菌种工厂化生产应用现状及发展浅析.食用菌,40(6):8-10
冀宏,赵黎明,2009.食用菌工厂化生产技术改进方法与思路.中国食用菌,28(6):56-58
Dai JQ,Zeng ZH,2012.Research status and development trend of liquid strains of edible fungi.Chinese Mushroom,2012(5):1-3(in Chinese)
Deacon J,2006.Fungal biology.Blackwell Publishing Ltd.,Oxford.1-366
Dong CH,Yao YJ,2010.Nutritional requirements of mycelial growth of Cordyceps sinensis in submerged culture.Journal of Applied Microbiology,99(3):483-492
Dubois M,Gilles K,Hamilton JK,Rebers PA,Smith F,1951.Colorimetric method for the determination of sugars.Nature,168(4265):167
Fang QH,Zhong JJ,2002.Effect of initial pH on production of ganoderic acid and polysaccharide by submerged fermentation of Ganoderma lucidum.Process Biochemistry,37(7):769-774
Francis F,Abdulhameed S,Madhavan N,Sumitra R,Ashok P,Sanjoy G,George S,2003.Use of response surface methodology for optimizing process parameters for the production of alpha-amylase by Aspergillus oryzae.Biochemical Engineering Journal,15(2):107-115
Fu YQ,Yin LF,Zhu HY,Jiang R,Li S,Xu Q,2014.Effects of pellet characteristics on L-lactic acid fermentation by R.oryzae:pellet morphology,diameter,density,and interior structure.Applied Biochemistry and Biotechnology,174(6):2019-2030
Ji H,Zhao LM,2009.Methods and ideas for improving industrial production technology of edible fungi.Chinese Mushroom,28(6):56-58(in Chinese)
Jin ZX,Wang YL,Wang PZ,Sun YB,Zhang ZS,2014.Optimization of culture condition for liquid fermentation of morchella.Journal of Dalian University of Technology,2014(6):416-419(in Chinese)
Kirsch LDS,Ana JPDM,Maria FST,2016.Production of mycelial biomass by the Amazonian edible mushroom Pleurotus albidus.Brazilian Journal of Microbiology,47(3):658-664
Li H,Zhang M,Li C,Zhang JJ,2018.Study on liquid culture technology of morels.Journal of Shenyang Agricultural University,2018(2):227-233(in Chinese)
Li JS,Ma CQ,Ma YH,Li Y,Zhou W,Xu P,2007.Medium optimization by combination of response surface methodology and desirability function:an application in glutamine production.Applied Microbiology and Biotechnology,74(3):563-571
Liu QY,Qi J,Zhou HY,Sun B,Wu HL,2018.Brief analysis on current situation and development of industrial production and application of edible fungus liquid strain.Journal of Dalian University of Technology,40(6):8-10(in Chinese)
Saad N,Abdeshahian P,Kalil MS,Yusoff WM,Hamid AA,2014.Optimization of aeration and agitation rate for lipid and gamma linolenic acid production by Cunninghamella bainieri 2A1 in submerged fermentation using response surface methodology.Scientific World Journal,2014(1):280146
Srinivas MRS,Nagin C,Lonsane BK,1994.Use of Plackett-Burman design for rapid screening of several nitrogen sources,growth/product promoters,minerals and enzyme inducers for the production of alpha-galactosidase by Aspergillus niger MRSS 234 in solid state fermentation system.Bioprocess Engineering,10(3):139-144
Vishvakarma R,Abha M,2017.Production of a protease inhibitor from edible mushroom Agaricus bisporus and its statistical optimization by response surface methodology.Preparative Biochemistry&Biotechnology,47(5):450-457
Wang D,Akiyoshi S,Motoyuki S,2001.Biological efficiency and nutritional value of Pleurotus ostreatus cultivated on spent beer grain.Bioresource Technology,78(3):293-300
Wu JZ,Peter CKC,Ka HW,Nian LH,2004.Studies on submerged fermentation of Pleurotus tuber-regium(Fr.)Singer.Part 2:effect of carbon-to-nitrogen ratio of the culture medium on the content and composition of the mycelial dietary fibre.Food Chemistry,85(1):101-105
Xiao JH,Dai XC,Wei HW,Xi JH,Ying Q,Zong QL,2006.Enhanced simultaneous production of mycelia and intracellular polysaccharide in submerged cultivation of Cordyceps jiangxiensis using desirability functions.Process Biochemistry,41(8):1887-1893
Zhang RY,Dan DH,Xiao TM,Shi GL,Jin GG,Qing XH,2014.Adopting stick spawn reduced the spawn running time and improved mushroom yield and biological efficiency of Pleurotus eryngii.Scientia Horticulturae,175(1):156-159
Zhu LW,Hong ML,Dong SL,Ya JT,2007.Submerged culture of mushrooms in bioreactors-challenges,current state-of-the-art,and future prospects.Food Technology&Biotechnology,45(3):221-229
戴建清,曾志恒,2012.食用菌液体菌种研究现状及发展趋势.中国食用菌,2012(5):1-3
李红,张敏,李超,张季军,2018.羊肚菌液体培养工艺研究.沈阳农业大学学报,2018(2):227-233
金朝霞,王云龙,王培忠,孙业兵,张宗申,2014.羊肚菌液体发酵培养条件优化.大连工业大学学报,2014(6):416-419
刘启燕,戚俊,周洪英,孙波,吴洪丽,2018.食用菌液体菌种工厂化生产应用现状及发展浅析.食用菌,40(6):8-10
冀宏,赵黎明,2009.食用菌工厂化生产技术改进方法与思路.中国食用菌,28(6):56-58