调控pH提高分批发酵赤霉素GA_3产量
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摘要
为高效率发酵生产GA3,对藤仓赤霉菌发酵过程pH进行优化调控研究。采用5L全自动发酵罐,在pH 3.0–5.0条件下,对藤仓赤霉菌菌丝生长及GA3产量的影响进行了考察,实验数据表明:在pH 4.0条件下,菌比生长速率可获最大值,为0.395/h;而pH 3.0条件下,GA3比生成速率最大,达到4.43mg/(g?h)。基于不同pH条件下,对菌比生长速率、得率、GA3比生成速率的影响,提出GA3分批发酵过程中的pH调控策略,即:0–20h,pH自然;20–50h,pH 4.0;50–80h,pH 3.0–3.5;80h后控制pH为3.5–4.0。在此控制模式下,经过196h发酵GA3的终产量达到2224mg/L,GA3产率44.5mg/g,GA3生产强度0.242mg/(L?h),分别比不控制pH条件下发酵的数值增长了7.75%、7.74%、8.04%,表明该pH控制策略能增进GA3发酵生产效率。
The effects of medium pH on the mycelium growth and GA3 production of Gibberella fujikuroi fermented in 5 L automatic bioreactor were investigated. The experiment results showed that under the condition of pH 4.0, the maximum growth rate of strain ZNL-13 was 0.395/h, while under pH 3.0 the GA3 production rate reached a maximum of 4.43 mg/(g?h). It was proposed that in batch fermentation the medium pH value should be regulated as follows on the basis of fermentation process: 0–20 h, natural pH; 20–50 h, pH 4.0;50–80 h, pH 3.0–3.5; over 80 h, pH 3.5–4.0. Under such a control mode, as a result of 196 h fermentation, GA3 final yield reached 2 224 mg/L, final productivity 44.5 mg/g and production capacity 0.242 mg/(L?h), 7.75%,7.74% and 8.04% higher than those under uncontrolled pH condition, respectively. It was indicated that GA3 production could be promoted by regulation of pH during Gibberella fujikuroi batch fermentation.
The effects of medium pH on the mycelium growth and GA3 production of Gibberella fujikuroi fermented in 5 L automatic bioreactor were investigated. The experiment results showed that under the condition of pH 4.0, the maximum growth rate of strain ZNL-13 was 0.395/h, while under pH 3.0 the GA3 production rate reached a maximum of 4.43 mg/(g?h). It was proposed that in batch fermentation the medium pH value should be regulated as follows on the basis of fermentation process: 0–20 h, natural pH; 20–50 h, pH 4.0;50–80 h, pH 3.0–3.5; over 80 h, pH 3.5–4.0. Under such a control mode, as a result of 196 h fermentation, GA3 final yield reached 2 224 mg/L, final productivity 44.5 mg/g and production capacity 0.242 mg/(L?h), 7.75%,7.74% and 8.04% higher than those under uncontrolled pH condition, respectively. It was indicated that GA3 production could be promoted by regulation of pH during Gibberella fujikuroi batch fermentation.
引文
Escamilla EM,Dendooven L,Maga?a IPR,De lTM,2000.Optimization of gibberellic acid production by immobilized Gibberella fujikuroi mycelium in fluidized bioreactors.Journal of Biotechnology,76(2):147-155
Gueguim-Kana EB,Oloke JK,Lateef A,Zebaze-Kana MG,2007.Novel optimal temperature profile for acidification process of Lactobacillus bulgaricus and Streptococcus thermophilus in yoghurt fermentation using artificial neural network and genetic algorithm.Journal of Industrial Microbiology and Biotechnology,34(7):491-496
Oller-López JL,Avalos J,Barrero AF,Oltra JE,2003.Improved GA1 production by Fusarium fujikuroi.Applied Microbiology and Biotechnology,63(3):282-285
Sun QX,Chen XS,Ren XD,Zheng GC,Mao ZG,2015.Enhancedε-poly-L-lysine production through pHregulation and organic nitrogen addition in fed-batch fermentation.Chinese Journal of Biotechnology,31(5):752-766(in Chinese)
Wang DM,Lv SX,Wang JS,2009.Guidance of biochemical experiment.Science Press,Beijing.103(in Chinese)
Wang W,Li JL,Huang WW,Li ZH,Zeng BQ,2014.Screening and identification of high gibberellin-producing strain from terbinafine resistant mutants.Microbiology China,42(9):1837-1842(in Chinese)
Wang W,Li ZH,Li JL,Zeng BQ,2011.Screening of high yield gibberellin-producing strain from lipase-producing mutants.Industrial Microbiology,41(1):51-55(in Chinese)
Wang W,Wu YH,Li JL,Yao YF,2017a.Enhanced gibberellic acid production through temperature regulation in batch fermentation.Chinese Journal of Applied&Environmental Biology,23(3):432-436(in Chinese)
Wang W,Wu YH,Li JL,Yao YF,2017b.Enhancement of gibberellin acid production through dissolved oxygen regulation in batch fermentation.Mycosystema,36(5):611-617(in Chinese)
Wu F,Yuan Y,He SH,Bandara AR,Hyde KD,Malysheva VF,Dai YC,2015.Global diversity and taxonomy of the Auricularia auricula-judae complex(Auriculariales,Basidiomycota).Mycological Progress,14:95-110
Wu J,Zhang KC,2006.Application of pH controlled strategy for enhancing exopolysaccharide production by Auricularia auricular.Chinese Journal of Applied&Environmental Biology,12(6):783-787(in Chinese)
Yan FG,He ZG,Qin J,Li JL,1995.Study on the fermentation of gibberellin A4,A7.Acta Mycologica Sinica,14(4):302-309(in Chinese)
Zhuang MK,2007.Breeding of gibberellic acid overproducting strain and fermentation.Master Thesis,Huazhong Agricultural University,Wuhan.1-63(in Chinese)
孙启星,陈旭升,任喜东,郑根成,毛忠贵,2015.基于pH调节和有机氮源流加调控补料分批发酵过程提高ε-聚赖氨酸产量.生物工程学报,31(5):752-766
王冬梅,吕淑霞,王金胜,2009.生物化学实验指导.北京:科学出版社.103
王卫,黎继烈,黄卫文,李忠海,曾柏全,2014.利用特比萘芬抗性筛选赤霉素高产菌株及相关发酵特性的研究.微生物学通报,41(9):1837-1842
王卫,李忠海,黎继烈,曾泊全,2011.以脂肪酶为指标筛选赤霉素高产菌株的研究.工业微生物,41(1):51-54
王卫,吴耀辉,黎继烈,姚跃飞,2017a.温度对赤霉素发酵的影响及优化策略.应用与环境生物学报,23(3):432-436
王卫,吴耀辉,黎继烈,姚跃飞,2017b.赤霉素发酵溶氧优化调控研究.菌物学报,36(5):611-617
吴芳,戴玉成,2015.黑木耳复合群中种类学名说明.菌物学报,34:604-611
吴静,章克昌,2006.基于动力学分析提高黑木耳多糖产量的pH控制策略.应用与环境生物学报,12(6):783-787
颜方贵,何增国,秦杰,李季伦,1995.赤霉素A4、A7的发酵研究.真菌学报,14(4):302-309
庄木坤,2007.赤霉素高产菌株改造与发酵研究.华中农业大学硕士论文,武汉.1-63
Gueguim-Kana EB,Oloke JK,Lateef A,Zebaze-Kana MG,2007.Novel optimal temperature profile for acidification process of Lactobacillus bulgaricus and Streptococcus thermophilus in yoghurt fermentation using artificial neural network and genetic algorithm.Journal of Industrial Microbiology and Biotechnology,34(7):491-496
Oller-López JL,Avalos J,Barrero AF,Oltra JE,2003.Improved GA1 production by Fusarium fujikuroi.Applied Microbiology and Biotechnology,63(3):282-285
Sun QX,Chen XS,Ren XD,Zheng GC,Mao ZG,2015.Enhancedε-poly-L-lysine production through pHregulation and organic nitrogen addition in fed-batch fermentation.Chinese Journal of Biotechnology,31(5):752-766(in Chinese)
Wang DM,Lv SX,Wang JS,2009.Guidance of biochemical experiment.Science Press,Beijing.103(in Chinese)
Wang W,Li JL,Huang WW,Li ZH,Zeng BQ,2014.Screening and identification of high gibberellin-producing strain from terbinafine resistant mutants.Microbiology China,42(9):1837-1842(in Chinese)
Wang W,Li ZH,Li JL,Zeng BQ,2011.Screening of high yield gibberellin-producing strain from lipase-producing mutants.Industrial Microbiology,41(1):51-55(in Chinese)
Wang W,Wu YH,Li JL,Yao YF,2017a.Enhanced gibberellic acid production through temperature regulation in batch fermentation.Chinese Journal of Applied&Environmental Biology,23(3):432-436(in Chinese)
Wang W,Wu YH,Li JL,Yao YF,2017b.Enhancement of gibberellin acid production through dissolved oxygen regulation in batch fermentation.Mycosystema,36(5):611-617(in Chinese)
Wu F,Yuan Y,He SH,Bandara AR,Hyde KD,Malysheva VF,Dai YC,2015.Global diversity and taxonomy of the Auricularia auricula-judae complex(Auriculariales,Basidiomycota).Mycological Progress,14:95-110
Wu J,Zhang KC,2006.Application of pH controlled strategy for enhancing exopolysaccharide production by Auricularia auricular.Chinese Journal of Applied&Environmental Biology,12(6):783-787(in Chinese)
Yan FG,He ZG,Qin J,Li JL,1995.Study on the fermentation of gibberellin A4,A7.Acta Mycologica Sinica,14(4):302-309(in Chinese)
Zhuang MK,2007.Breeding of gibberellic acid overproducting strain and fermentation.Master Thesis,Huazhong Agricultural University,Wuhan.1-63(in Chinese)
孙启星,陈旭升,任喜东,郑根成,毛忠贵,2015.基于pH调节和有机氮源流加调控补料分批发酵过程提高ε-聚赖氨酸产量.生物工程学报,31(5):752-766
王冬梅,吕淑霞,王金胜,2009.生物化学实验指导.北京:科学出版社.103
王卫,黎继烈,黄卫文,李忠海,曾柏全,2014.利用特比萘芬抗性筛选赤霉素高产菌株及相关发酵特性的研究.微生物学通报,41(9):1837-1842
王卫,李忠海,黎继烈,曾泊全,2011.以脂肪酶为指标筛选赤霉素高产菌株的研究.工业微生物,41(1):51-54
王卫,吴耀辉,黎继烈,姚跃飞,2017a.温度对赤霉素发酵的影响及优化策略.应用与环境生物学报,23(3):432-436
王卫,吴耀辉,黎继烈,姚跃飞,2017b.赤霉素发酵溶氧优化调控研究.菌物学报,36(5):611-617
吴芳,戴玉成,2015.黑木耳复合群中种类学名说明.菌物学报,34:604-611
吴静,章克昌,2006.基于动力学分析提高黑木耳多糖产量的pH控制策略.应用与环境生物学报,12(6):783-787
颜方贵,何增国,秦杰,李季伦,1995.赤霉素A4、A7的发酵研究.真菌学报,14(4):302-309
庄木坤,2007.赤霉素高产菌株改造与发酵研究.华中农业大学硕士论文,武汉.1-63