浮游球衣菌S9产铁氧化酶发酵条件优化及酶学特性研究
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摘要
通过单因素及正交设计试验,研究了浮游球衣菌S9产铁氧化酶的发酵条件,并探讨了铁氧化酶的酶学性质。结果表明,菌株S9的最适产酶发酵培养基配方为柠檬酸铁铵1.0%,NH_4Cl 0.1%,K_2HPO_40.05%,Mg SO_4·7H_2O 0.05%,Ca Cl_2·6H_2O 0.01%;最佳发酵条件为:培养初始p H7.0,培养温度30℃,装液量50 m L/250 m L,转速150 r/min,培养时间84 h。在此优化发酵条件下,菌株S9的铁氧化率可达75.25%,与未优化前相比,提高了42.28%。所产铁氧化酶的最适温度为30℃,最适p H为7.5,Mg~(2+)、K~+、Na~+对酶的活性具有增强作用,Pb~(2+)、Ag~+会抑制酶的部分活性,且Mn~(2+)、Zn~(2+)对酶具有很强的抑制作用。对该菌最适培养基和产酶条件及酶学性质,对铁氧化酶在给排水工程及其分离纯化和活性保护具有一定的参考价值和指导意义。
Fermentation conditions of Sphaerotilus natans S9 for ferroxidase production were studied by single factor and orthogonal experiments,and the enzymatic properties of ferroxidase were studied.The results showed that the optimum fermentation medium of strain S9 was ferric ammonium citrate 1.0%,NH_4Cl 0.1%,K_2HPO_40.05%,Mg SO_4·7H_2O 0.05%,Ca Cl_2·6H_2O 0.01%.The optimum fermentation conditions were as follows:initial p H 7.0,culture temperature 30℃,liquid volume 50 ml/250 ml,rotational speed 150 r/min,culture time 84 h.Under the fermentation conditions,the ferro-oxidation rate of strain S9 can reach 75.25%,and ferro-oxidation rate increased 42.28%compared to that of before optimization.The optimum temperature and p H of ferroxidase were 30℃and 7.5,respectively.Mg~(2+),K~+,Na~+had certain activation on ferroxidase,Pb~(2+),Ag~+had inhibitory effect on the enzyme activity,while Mn~(2+),Zn~(2+)had very strong inhibition on ferroxidase activity.The study on the optimum culture medium,enzyme production conditions and enzymatic properties was of certain reference value and guiding significance for the purification and protection of the enzyme in water supply and drainage engineering.
Fermentation conditions of Sphaerotilus natans S9 for ferroxidase production were studied by single factor and orthogonal experiments,and the enzymatic properties of ferroxidase were studied.The results showed that the optimum fermentation medium of strain S9 was ferric ammonium citrate 1.0%,NH_4Cl 0.1%,K_2HPO_40.05%,Mg SO_4·7H_2O 0.05%,Ca Cl_2·6H_2O 0.01%.The optimum fermentation conditions were as follows:initial p H 7.0,culture temperature 30℃,liquid volume 50 ml/250 ml,rotational speed 150 r/min,culture time 84 h.Under the fermentation conditions,the ferro-oxidation rate of strain S9 can reach 75.25%,and ferro-oxidation rate increased 42.28%compared to that of before optimization.The optimum temperature and p H of ferroxidase were 30℃and 7.5,respectively.Mg~(2+),K~+,Na~+had certain activation on ferroxidase,Pb~(2+),Ag~+had inhibitory effect on the enzyme activity,while Mn~(2+),Zn~(2+)had very strong inhibition on ferroxidase activity.The study on the optimum culture medium,enzyme production conditions and enzymatic properties was of certain reference value and guiding significance for the purification and protection of the enzyme in water supply and drainage engineering.
引文
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[18]秦玉春,关晓辉,魏德洲,等.浮游球衣菌的分离及其对铅离子的吸附性能[J].东北大学学报:自然科学版,2005,26(7):687-690.
[2]秦玉春,秦玉华.浮游球衣菌-纳米Fe3O4生物吸附剂处理电镀废水中Cd(Ⅱ)和Cr(Ⅵ)的研究[J].广东化工,2014,41(8):108,103.
[3]郑虹,乐增焜,邓加聪.1株鞘细菌的分离筛选及对铅离子吸附条件的研究[J].微生物学杂志,2014,34(2):43-48.
[4]CHOI J D,KOTAY S M,GOEL R.Bacteriophage-based biocontrol of biological sludge bulking in waster[J].Bioengineered Bugs,2011,2(4):214-217.
[5]SEDER-COLOMINA M,MORIN G,BENZERARA K,et al.Sphaerotilus natans,a neutrophilic iron-related sheath-forming bacteriaum:perspectires for metal remediation stategies[J].Geomicrob J,2013,31(1):64-75.
[6]郑虹,崔伟斌,林廉晶,等.鞘细菌的生物包埋及固定化吸附条件的研究[J].福建师大福清分校学报,2016,135(2):33-41.
[7]佘晨兴,许旭萍,林跃鑫,等.鞘细菌FC9901氧化铁生化机制的研究[J].环境污染治理技术与设备,2002,3(9):35-37.
[8]郭艳.富集重金属鞘细菌的分离筛选、吸附条件及固定化研究[D].福州:福建师范大学,2008:1-63.
[9]KAMPFER P.Some chemotaxonomic and physiological properties of the genus Sphaerotilus[J].Syst Appl Microbiol,1998,21(2):245-250.
[10]BOSSIS F,PALESE L.Molecular dynamics in cytochrome c-oxidase mossbauer spectra deconvolution[J].Biochem Biophy Res Comm,2011,404(17):438-442.
[11]许旭萍,杨冠彬,王芳,等.假单胞菌产生铁氧化酶条件的研究[J].微生物学杂志,2009,29(4):16-19.
[12]江令翔,张翔宇,朱秀玲,等.活性污泥中氧化铁细菌的分离鉴定及酶活的初步研究[J].安徽工程大学学报,2015,30(5):13-18.
[13]邓加聪,王诗瑶,蒋素素,等.高效产锰氧化酶鞘细菌的分离鉴定及酶学特性研究[J].中国酿造,2017,36(10):97-103.
[14]JABORNING S,PODMIRSEG S M.A novel fixed fibre biofilm membrane process for on-site greywater reclamation requiring no fouling control[J].Biotechnol Bioeng,2015,112(3):484-493.
[15]郑虹,张晓榕,吴晓梅,等.高效铁氧化鞘细菌的筛选与鉴定[J].微生物学报,2014,54(12):1352-1358.
[16]国家环境保护总局,水和废水监测分析方法编委会.水和废水监测分析方法(第四版)[M].北京:中国环境科学出版社,2002:368-370.
[17]美国公共卫生协会,宋仁元,张亚杰,等译.水和废水标准检验法(第十五版)[M].北京:中国建筑工业出版社,1985:180-186.
[18]秦玉春,关晓辉,魏德洲,等.浮游球衣菌的分离及其对铅离子的吸附性能[J].东北大学学报:自然科学版,2005,26(7):687-690.