枯草芽孢杆菌产过氧化氢酶的优化与工业化
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摘要
为了实现过氧化氢酶的工业化生产,作者对实验室前期构造成功的B.subtilis WSHDZ-01(p STOP1622-kat A)进行了3 L发酵罐的发酵验证,并且通过数据分析优化了碳氮源浓度,使得过氧化氢酶酶活达到35 398 U/m L,比优化前提高53.41%。在此基础上,分别进行了24 L发酵罐和500 L发酵罐的放大实验,最高酶活分别达到28 940、23 190 U/m L。结果表明,该过氧化氢酶在产酶水平上已经达到工业化生产需求,为更大规模的工业化生产奠定坚实的基础。
Microorganism catalase,which is an important industrial enzyme,plays an important role in textile industry. To realize the industrial production of catalase,the fermentation on B. subtilis WSHDZ-01(p STOP1622-kat A),which was constructed before,was conducted in 3 L fermentor.Furthermore,the enzyme activity reached 35 398 U/m L after optimization of carbon and nitrogen concentration,which was increased by 53.41% compared with that before optimization. Based on this result,the fermentations in 24 L and 500 L fermentor were carried out,respectively and the maximum enzyme activity was 28 940 U/m L and 23 190 U/m L respectively. The result showed the yield of catalase has met the requirement of industrial production,which lay a solid foundation for larger-scale industrial production.
Microorganism catalase,which is an important industrial enzyme,plays an important role in textile industry. To realize the industrial production of catalase,the fermentation on B. subtilis WSHDZ-01(p STOP1622-kat A),which was constructed before,was conducted in 3 L fermentor.Furthermore,the enzyme activity reached 35 398 U/m L after optimization of carbon and nitrogen concentration,which was increased by 53.41% compared with that before optimization. Based on this result,the fermentations in 24 L and 500 L fermentor were carried out,respectively and the maximum enzyme activity was 28 940 U/m L and 23 190 U/m L respectively. The result showed the yield of catalase has met the requirement of industrial production,which lay a solid foundation for larger-scale industrial production.
引文
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[2]Cabiscol E,Tamarit J,Ros J.Oxidative stress in bacteria and protein damage by reactive oxygen species[J].International Microbiology,2010,3(1):3-8.
[3]Usui S,Komeima K,Lee S Y,et al.Increased expression of catalase and superoxide dismutase 2 reduces cone cell death in retinitis pigmentosa[J].Molecular Therapy,2009,17(5):778-786.
[4]Gudelj M,Fruhwirth G,Paar A,et al.A catalase-peroxidase from a newly isolated thermoalkaliphilic Bacillus sp.with potential for the treatment of textile bleaching effluents[J].Extremophiles,2001,5(6):423-429.
[5]冷晒祥,钱国坻,华兆哲,等.过氧化氢酶的棉针织物漂染工艺研究[J].印染,2006,32(19):1-3.LENG Shaixiang,QIAN Guodi,GUA Zhaozhe,et al.Catalase for H2O2removal after cotton knits bleaching[J].Dyeing&Finishing,2006,32(19):1-3.(in Chinese)
[6]张东旭,堵国成,陈坚.微生物过氧化氢酶的发酵生产及其在纺织工业的应用[J].生物工程学报,2010,26(11):1473-1481.ZHANG Dongxu,DU Guocheng,CHEN Jian.Fermentation production of microbial catalase and its application in textile industry[J].Chinese Journal of Biotechnology,2010,26(11):1473-1481.(in Chinese)
[7]姚丹丹,刘立明,李江华,等.活性氧胁迫促进枯草芽孢杆菌WSHDZ-01过量合成过氧化氢酶[J].生物工程学报,2009,25(5):786-792.YAO Dandan,LIU Liming,LI Jianghua,et al.Overproduction of catalove by oxidative stress on Bacillus subtilis WSHDZ-01[J].Chinese Journal of Biotechnology,2009,25(5):786-792.(in Chinese)
[8]Biedendieck R,Yang Y,Deckwer WD,et al.Plasmid system for the intracellular production and purification of affinity-tagged proteins in Bacillus megaterium[J].Biotechnology and Bioengineering,2007,96(3):525-537.
[9]Dwivedi P,Wu P,Klopsch S J,et al.Metabolic profiling by ion mobility mass spectrometry(IMMS)[J].Metabolomics,2008,4(1):63-80.
[10]Zhu S,Wu Y,Yu Z,et al.Production of ethanol from microwave-assisted alkali pretreated wheat straw[J].Process Biochemistry,2006,41(4):869-873.
[11]Rai U,Sinha S,Tripathi R,et al.Wastewater treatability potential of some aquatic macrophytes:removal of heavy metals[J].Ecological Engineering,1995,5(1):5-12.