海洋尿酸氧化酶菌株发酵条件响应面优化
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摘要
在单因素试验基础上,采用Plackett-Burman试验得出尿酸、酵母膏、温度是3个最重要的影响菌株产酶的因素,利用BoxBehnken模型对苛求芽孢杆菌(Bacillus fastidious)Z7-1发酵生产尿酸氧化酶的产酶条件进行响应面优化。单因素优化结果为尿酸1.00%、酵母膏0.75%、K2HPO40.25%、Mg SO40.05%、KH2PO40.07%、培养温度25℃、转速160 r/min、接种量5%、p H 7.5、装液量125 m L/500 m L。响应面优化后发酵条件为尿酸1.00%、酵母膏0.80%和温度28℃。在此优化条件下,酶活为42.57 U/m L,比优化前提高了251.8%。
On the basis of single factor experiment, three most important factors affecting the enyme production of strains were determined as uric acid,yeast extract and temperature by Plackett-Burman test. Then the urate oxidase producing conditions of Bacillus fastidious Z7-1 was optimized by Box-Behnken model and response surface methodology. The single factor optimization results were uric acid 1.00%, yeast extract 0.75%, K2 HPO40.25%, Mg SO40.05%, KH2 PO40.07%, culture temperature 25 ℃, rotation speed 160 r/min, inoculum 5%, p H 7.5, liquid volume 125 ml/500 ml. After the response surface optimization, the fermentation conditions were optimized as uric acid 1.00%, yeast extract 0.80% and temperature 28 ℃. Under the optimized conditions, the enzyme activity was 42.57 U/ml, which was 251.8% higher than before optimization.
On the basis of single factor experiment, three most important factors affecting the enyme production of strains were determined as uric acid,yeast extract and temperature by Plackett-Burman test. Then the urate oxidase producing conditions of Bacillus fastidious Z7-1 was optimized by Box-Behnken model and response surface methodology. The single factor optimization results were uric acid 1.00%, yeast extract 0.75%, K2 HPO40.25%, Mg SO40.05%, KH2 PO40.07%, culture temperature 25 ℃, rotation speed 160 r/min, inoculum 5%, p H 7.5, liquid volume 125 ml/500 ml. After the response surface optimization, the fermentation conditions were optimized as uric acid 1.00%, yeast extract 0.80% and temperature 28 ℃. Under the optimized conditions, the enzyme activity was 42.57 U/ml, which was 251.8% higher than before optimization.
引文
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[12]孙子羽,迟乃玉,李兵,等.响应面法优化产低温生淀粉糖化酶发酵培养基[J].中国酿造,2010,29(7):110-114.
[13]齐西珍,王利强,孟鹏,等.利用响应面法优化α-糖苷酶抑制剂发酵培养基[J].微生物学通报,2012,39(2):203-210.
[14]袁慎亮,邢德明,窦少华,等.高产纤溶酶菌株CNY16发酵条件优化及其酶学特性初步研究[J].微生物学通报,2014,41(8):1621-1628.
[15]王春雨,迟乃玉,张庆芳,等.产低温脂肪酶菌株CZW001发酵培养基优化研究[J].微生物学通报,2013,40(9):1541-1549.
[16]蒋彪,王常高,杜馨,等.响应面法优化芽孢杆菌CJPE209产角蛋白酶发酵培养基的研究[J].中国酿造,2017,36(5):76-80.
[17]杨梦晨,户红通,刘子强,等.响应面分析优化L-色氨酸清液发酵培养基[J].生物资源,2017(6):464-471.
[18]周新尚,逢飞,窦少华,等.海洋生淀粉酶菌株发酵条件响应面优化[J].中国酿造,2017,36(7):80-84.
[2]YAMAMOTO K,KOJIMA Y,KIKUCHI T,et al.Nucleotide sequence of the uricase gene from Bacillus sp.TB-90[J].J Biochem,1996,119(1):80-84.
[3]MONTALBINI P,REDONDO J,CABALLERO J L,et al.Uricase from leaves:its purification and characterization from three different higher plants[J].Planta,1997,202(3):277-283.
[4]ABDEL-FATTAH Y R,SAEED H M,GOHAR Y M,et al.Improved production of Pseudomonas aeruginosa uricase by optimization of process parameters through statistical experimental designs[J].Process Biochem,2005,40(5):1707-1714.
[5]SCHIAVON O,CALICETI P,FERRUTI P,et al.Therapeutic proteins:a comparison of chemical and biological properties of uricase conjugated to linear or branched poly(ethylene glycol)and poly(N-acryloylmorpholine)[J].IL Farmaco,2000,55(4):264-269.
[6]李文杰.枯草芽孢杆菌尿酸氧化酶的定向进化[D].合肥:中国科学技术大学,2017.
[7]陈志禹,何秀萍,张博润.微生物来源的尿酸氧化酶的研究进展及应用前景[J].微生物学通报,2007,34(6):1205-1208.
[8]BONGAERTS G P,UITZETTER J,BROUNS R,et al.Uricase of Bacillus fastidiosus.properties and regulation of synthesis[J].BBA-Enzymology,1978,527(2):348-358.
[9]GHOSH T,SARKAR P.Isolation of a novel uric-acid-degrading microbe Comamonas sp.BT UA and rapid biosensing of uric acid from extracted uricase enzyme[J].J Biosci,2014,39(5):805-819.
[10]HUANG S H,WU T K.Modified colorimetric assay for uricase activity and a screen for mutant Bacillus subtilis uricase genes following St EP mutagenesis[J].Febs J,2004,271(3):517-523.
[11]张玉然,纪楠楠.诊断用耐热尿酸氧化酶菌株的筛选及条件优化[J].中国生化药物杂志,2016,36(9):154-157.
[12]孙子羽,迟乃玉,李兵,等.响应面法优化产低温生淀粉糖化酶发酵培养基[J].中国酿造,2010,29(7):110-114.
[13]齐西珍,王利强,孟鹏,等.利用响应面法优化α-糖苷酶抑制剂发酵培养基[J].微生物学通报,2012,39(2):203-210.
[14]袁慎亮,邢德明,窦少华,等.高产纤溶酶菌株CNY16发酵条件优化及其酶学特性初步研究[J].微生物学通报,2014,41(8):1621-1628.
[15]王春雨,迟乃玉,张庆芳,等.产低温脂肪酶菌株CZW001发酵培养基优化研究[J].微生物学通报,2013,40(9):1541-1549.
[16]蒋彪,王常高,杜馨,等.响应面法优化芽孢杆菌CJPE209产角蛋白酶发酵培养基的研究[J].中国酿造,2017,36(5):76-80.
[17]杨梦晨,户红通,刘子强,等.响应面分析优化L-色氨酸清液发酵培养基[J].生物资源,2017(6):464-471.
[18]周新尚,逢飞,窦少华,等.海洋生淀粉酶菌株发酵条件响应面优化[J].中国酿造,2017,36(7):80-84.
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