(+)γ-内酰胺酶菌株BH24发酵产酶条件优化
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摘要
基于单因素试验,通过Plackett-Burman试验设计、最陡爬坡试验设计、中心组合试验设计和响应面分析,对(+)γ-内酰胺酶菌株BH24发酵产酶进行优化。单因素试验结果表明,在碳源为5.0 g/L葡萄糖、氮源为10.0 g/L蛋白胨、温度26.0℃、pH 8.0、接种量4.0%、装液量100 mL/500 m L、转速150 r/min条件下,菌株BH24的生物量、(+)γ-内酰胺酶酶活力及产物e.e.值均有不同程度的提高。响应面优化结果表明,当葡萄糖质量浓度为4.0 g/L,温度为25.2℃,pH值为7.6时,(+)γ-内酰胺酶酶活可达114.5 U/L,比优化前提高了50.7%。
Based on single factor experiments, the fermentation conditions of (+) γ-lactamases-producting strain BH24 were optimized by Plackett Burman design, the steepest ascent experiments design, central composite design, and response surface analysis. The results of single factor experiments showed that in the conditions of 5.0 g/L glucose as carbon source, 10.0 g/L peptone as the nitrogen source, temperature 26.0 ℃, pH 8.0, inoculum 4.0%, liquid volume 100 ml/500 ml, rotating speed 150 r/min, the biomass,(+) γ-lactamases activity and e.e. value all were increased in different degrees. The results of response surface optimization showed that the (+) γ-lactamases activity was up to 114.5 U/L in the conditions of glucose content 4.0 g/L, temperature 25.2 ℃ and pH 7.6, which was 50.7% higher than that before optimization
Based on single factor experiments, the fermentation conditions of (+) γ-lactamases-producting strain BH24 were optimized by Plackett Burman design, the steepest ascent experiments design, central composite design, and response surface analysis. The results of single factor experiments showed that in the conditions of 5.0 g/L glucose as carbon source, 10.0 g/L peptone as the nitrogen source, temperature 26.0 ℃, pH 8.0, inoculum 4.0%, liquid volume 100 ml/500 ml, rotating speed 150 r/min, the biomass,(+) γ-lactamases activity and e.e. value all were increased in different degrees. The results of response surface optimization showed that the (+) γ-lactamases activity was up to 114.5 U/L in the conditions of glucose content 4.0 g/L, temperature 25.2 ℃ and pH 7.6, which was 50.7% higher than that before optimization
引文
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[9]吴伟,曾倩兰,向福.响应面分析法优化火棘果中原花青素提取工艺[J].中国酿造,2015,34(10):116-120.
[10]武顺,王德良,闫寅卓.产吡嗪类细菌的麸曲制作工艺优化[J].中国酿造,2016,35(2):13-17.
[11]韩健,刘朝辉,齐崴,等.β-甘露聚糖酶发酵液絮凝条件的统计学筛选与响应面优化[J].生物加工过程,2007,5(2):29-35.
[12]赵世光,王诗然,薛正莲,等.Ganoderma lucidum UIM-281产漆酶发酵条件的筛选及响应面优化[J].食品与发酵工业,2009,35(11):72-77.
[13]王颖,何禾,胡发根,等.响应面法优化重组大肠杆菌全细胞合成D-苯基乳酸[J].食品科学,2016,37(7):88-92.
[14]刘京兰,蔡勋超,薛雅蓉,等.生防解淀粉芽孢杆菌CC09合成iturin A条件的响应面优化[J].中国生物防治学报,2016(2):235-243.
[15]周生民,冯文红,赵伟.响应面法优化黑曲霉发酵产葡萄糖酸钙[J].生物加工过程,2016(2):17-21.
[2]BRABBAN A D,LITTLECHILD J A,WISDOM R.Stereospecificγ-lactamase activity in a Pseudomonas fluorescens species[J].J Ind Microbiol,1996,16(1):8-14.
[3]CHAND P,KOTIAN P L,DEHGHANI A,et al.Systematic structure based design and stereoselective synthesis of novel multisubstitu tedcyclopentane derivatives with potent antiinfluenzaactivity[J].J Med Chem,2001,44(25):4379-4392.
[4]LAGOJA I M,CLERCQ E D.Anti-influenzavirus agents:synthesis and mode of action[J].Med Res Rev,2008,28(1):1-38.
[5]TOOGOOD H S,BROWN R C,LINE K,et al.The use of a thermostable signature amidase in the resolution of the bicyclic synthon(rac)γ-lactam[J].Tetrahedron,2004,60(3):711-716.
[6]TAYLOR S J C,SUTHERLAND A G,LEE C,et al.Chemoenzymatic synthesis of(-)-carbovir utilizing a whole cell catalysed resolution of2-azabicyclo[2.2.2]hept-5-en-3-one[J].J Chem Soc:Chem Commun,1990,16(1):1120-1121.
[7]GIORDANO C,AMMENDOLA S.Characterization of mutants of Sulfolobus solfataricus signature amidase able to hydrolyse R-ketoprofen amide[J].Protein Peptide Lett,2008,15(6):617-623.
[8]王建军,郑国钧,吴胜.微生物来源的γ-内酰胺酶的研究进展[J].微生物学报,2010,50(8):988-994.
[9]吴伟,曾倩兰,向福.响应面分析法优化火棘果中原花青素提取工艺[J].中国酿造,2015,34(10):116-120.
[10]武顺,王德良,闫寅卓.产吡嗪类细菌的麸曲制作工艺优化[J].中国酿造,2016,35(2):13-17.
[11]韩健,刘朝辉,齐崴,等.β-甘露聚糖酶发酵液絮凝条件的统计学筛选与响应面优化[J].生物加工过程,2007,5(2):29-35.
[12]赵世光,王诗然,薛正莲,等.Ganoderma lucidum UIM-281产漆酶发酵条件的筛选及响应面优化[J].食品与发酵工业,2009,35(11):72-77.
[13]王颖,何禾,胡发根,等.响应面法优化重组大肠杆菌全细胞合成D-苯基乳酸[J].食品科学,2016,37(7):88-92.
[14]刘京兰,蔡勋超,薛雅蓉,等.生防解淀粉芽孢杆菌CC09合成iturin A条件的响应面优化[J].中国生物防治学报,2016(2):235-243.
[15]周生民,冯文红,赵伟.响应面法优化黑曲霉发酵产葡萄糖酸钙[J].生物加工过程,2016(2):17-21.