响应面法优化生物活菌剂解淀粉芽孢杆菌JT-84发酵工艺
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摘要
通过优化解淀粉芽孢杆菌JT-84的发酵工艺,为其产业化生产提供理论依据。以解淀粉芽孢杆菌JT-84菌体生物量为指标,进行碳源(葡萄糖、麦芽糖、蔗糖、甘露醇、可溶性淀粉和乳糖)、氮源[(NH4) 2SO4、尿素、NaNO3、酵母粉、蛋白胨和牛肉膏]和无机盐(CuSO4、MgSO4、NaCl、ZnCl2、CaCl2和KCl)的单因素试验,并筛选出9个主要影响因素;然后,通过PB试验和响应面分析等方法,对其发酵培养基配方进行优化,最终得到最佳发酵培养基配方为葡萄糖17.51 g/L、酵母粉9.88 g/L和CaCl22.11 g/L。在优化发酵培养基的基础上,以菌体生物量最大为标准,对发酵条件进行优化,得到最优发酵条件为菌体接种量8%、培养基初始pH值6.5、发酵温度28℃、摇瓶转速140 r/min、发酵时间48 h。
Optimizing the fermentation process of Bacillus amyloliquefaciens JT-84 can provide a theoretical basis for its industrial production. Using the bacterial biomass of B. amyloliquefaciens JT-84 as an index,the single factor experiments on carbon sources(glucose, maltose, sucrose, mannitol, soluble starch and lactose),nitrogen sources [(NH4)2SO4,urea,NaNO3,yeast powder, peptone and beef extract] and inorganic salts(CuSO4,MgSO4,NaCl,Zn Cl2,CaCl2 and KCl) were conducted,and nine major influencing factors were screened out. The fermentation medium formulation was optimized by PB experiment and response surface analysis. The optimal fermentation medium formulation was finally obtained with glucose 17.51 g/L,yeast powder 9.88 g/L and CaCl22.11 g/L. On the basis of optimizing the fermentation medium,the fermentation conditions were optimized based on the maximum bacterial biomass. The optimal fermentation conditions were 8% inoculation, initial p H value 6.5,fermentation temperature 28 ℃,shaker speed 140 r/min,and fermentation time 48 h.
Optimizing the fermentation process of Bacillus amyloliquefaciens JT-84 can provide a theoretical basis for its industrial production. Using the bacterial biomass of B. amyloliquefaciens JT-84 as an index,the single factor experiments on carbon sources(glucose, maltose, sucrose, mannitol, soluble starch and lactose),nitrogen sources [(NH4)2SO4,urea,NaNO3,yeast powder, peptone and beef extract] and inorganic salts(CuSO4,MgSO4,NaCl,Zn Cl2,CaCl2 and KCl) were conducted,and nine major influencing factors were screened out. The fermentation medium formulation was optimized by PB experiment and response surface analysis. The optimal fermentation medium formulation was finally obtained with glucose 17.51 g/L,yeast powder 9.88 g/L and CaCl22.11 g/L. On the basis of optimizing the fermentation medium,the fermentation conditions were optimized based on the maximum bacterial biomass. The optimal fermentation conditions were 8% inoculation, initial p H value 6.5,fermentation temperature 28 ℃,shaker speed 140 r/min,and fermentation time 48 h.
引文
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[19]邓建良,刘红彦,刘玉霞,刘新涛,倪云霞,高素霞,李国庆.解淀粉芽孢杆菌YN-1抑制植物病原真菌活性物质鉴定[J].植物病理学报,2010,40(2):202-209.
[20] Chen X H,Koumoutsia A,Scholza R,Schneider K,Vater J,Süssmuth R,Piel J,Borriss R. Genome analysis of Bacillus amyloliquefaciens FZB42 reveals itspotential for biocontrol of plant pathogens[J]. Journal of Biotechnology,2009,140(1-2):27-37.
[2]吴一晶,林艺芬,林河通,陈艺晖,陈梦茵,姜萍.生防菌解淀粉芽孢杆菌研究进展[J].包装与食品机械,2012,30(6):49-52.
[3]陈哲,黄静,赵佳,王长彪,梁宏.解淀粉芽孢杆菌抑菌机制的研究进展[J].生物技术通报,2015,31(6):37-41.
[4]侯美如,刘宇,王岩,尹珺伊,周庆民,秦平伟,史同瑞,杨淑萍.解淀粉芽胞杆菌固态发酵黄芪培养基的研究[J].中国兽医科学,2016,46(10):1328-1334.
[5] Wang X, Wang L, Li X, Xu Y. Response surface methodology based optimization for degradation of align in Laminaria japonica,feedstuff via fermentation by Bacillus,in Apostichopus japonicas,farming[J]. Electronic Journal of Biotechnology,2016,22:1-8.
[6]张丽姣,曾艳,张颖,宋诙,贾士儒,孙媛霞.解淀粉芽孢杆菌胞外多糖发酵条件的优化[J].食品工业科技,2015,36(9):214-219.
[7] Zhang W,Zhang X,Cui H. Isolation,fermentation optimization and performance studies of a novel biosurfactant producing strain Bacil lus amyloliquefaciens[J]. Chemical&Biochemical Engineering Quarterly, 2015, 29(3):447-456.
[8] Yan Z, Qun W, Yan X. Production of surfactin from waste distillers’ grains by co-culture fermentation of two Bacillus amyloliquefaciens strains[J]. Bioresource Technology,2017,235:96-103.
[9]章小洪,汪琨,朱廷恒,崔志峰.解淀粉芽孢杆菌BW-13培养基和培养条件优化[J].浙江工业大学学报,2013,41(1):35-39.
[10]李娟,夏凯丽,王远宏,董蕴琪,郝蕾.解淀粉芽孢杆菌LJ1摇瓶发酵条件优化[J].生物技术通报,2015,31(12):214-220.
[11]韩玉竹,邓钊,张宝,游成真,丛苑,李平兰.解淀粉芽孢杆菌H15产抗菌肽的发酵条件优化和提取方法比较研究[J].食品科学,2015,36(15):135-141.
[12]洪鹏,安国栋,胡美英,耿鹏,桑松.解淀粉芽孢杆菌HF-01发酵条件优化[J].中国生物防治学报,2013,29(4):569-578.
[13]黄海婵,裘娟萍.枯草芽孢杆菌防治植物病害的研究进展[J].浙江农业科学,2005,(3):213-215,219.
[14]牛力轩,王楠,王雪梅,胡江春,王书锦.一株产脂肽类抗生素bacillopeptin A深海芽孢杆菌的筛选与鉴定[J].中国抗生素杂志,2011,36(10):738-744,750.
[15]朱丽梅,吴小芹,胡飞杰,徐浩.解淀粉芽孢杆菌JK-JS3杀线活性物质的分离与结构分析[J].林业科学,2011,47(7):133-137.
[16]朱震,罗毅,张鹏,杨兴明,冉炜,沈其荣.产表面活性素和伊枯草菌素A菌株的筛选及其脂肽类产物的特性[J].微生物学通报,2011,38(10):1488-1498.
[17]薛鹏琦,刘芳,乔俊卿,伍辉军,冯致科,高学文.油菜菌核病生防芽孢杆菌的分离鉴定及其脂肽化合物分析[J].植物保护学报,2011,38(2):127-132.
[18]石磊,郭庆港,李宝庆,鹿秀云,李社增,王洪港,马平.作物病害高效生防菌株的定向快速筛选及其种类鉴定[J].安徽农业科学,2012,40(24):12068-12071,12096.
[19]邓建良,刘红彦,刘玉霞,刘新涛,倪云霞,高素霞,李国庆.解淀粉芽孢杆菌YN-1抑制植物病原真菌活性物质鉴定[J].植物病理学报,2010,40(2):202-209.
[20] Chen X H,Koumoutsia A,Scholza R,Schneider K,Vater J,Süssmuth R,Piel J,Borriss R. Genome analysis of Bacillus amyloliquefaciens FZB42 reveals itspotential for biocontrol of plant pathogens[J]. Journal of Biotechnology,2009,140(1-2):27-37.