氧化铁鞘细菌铁氧化酶最适产酶条件及其酶学特性的研究
STUDY ON THE OPTIMUM CULTURAL CONDITIONS FOR Fe2+ OXIDASE PRODUCTION OF Fe2+OXIDIZED SHEATHED BACTERIA SPHAEROTILUS NATANS FC9901 AND SOME ENZYMATIC CHARACTERISTICS OF Fe2+OXIDASE
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摘要: 对氧化铁鞘细菌FC9901菌株的铁氧化酶最适产酶条件及酶学特性进行了研究。菌株最适产酶培养基为(g/L):柠檬酸铁胺10g,NaNO31.2g,MgSO4·7H2O0.5g,K2HPO4·7H2O0.5g,CaCl2 0.015g,ZnSO4 7H2O0.0005 g。最适产酶条件为:温度30℃,起始pH7.0,接种量2%,150mL三角瓶装50mL,150r/min振荡培养72h。铁氧化酶最适pH为7.5,最适温度为30℃。金属离子Ca2+、Mg2+、Zn2+对酶有激活和稳定作用;Cu2+、Hg2+、Al3+则抑制酶的活性;Fe2+、K+、Na+对酶活性影响不明显。Abstract: Fe 2+-Mn2+oxidizing sheathed bacteria is a group of bacteria with ability to oxidize Fe2+ and Mn2+.In practice they were used to treat wastewater.Previous researches indicated that the functional component for Fe 2+-Mn2+ oxidation in the bacterium is Fe2+oxidase.Because of the low content and easy inactivation,Fe2+oxidase is difficult to be purified.This resulted the less information about the properties of the enzyme.A strain of Fe2+ oxidizing sheathed bacteria Sphaerotilus natans FC9901 we separated previous was used as material for Fe2+oxidase investigation in this paper.Serial culture media were employed to culture the bacterium so as to explore the optimum conditions for enzyme productivity.The modified TMPD method was used to measure the enzymatic activity.After partially purification by ammonia sulfate precipitation and dialysis,the enzymatic properties were analyzed.The results were as follows:the optimum culture medium for enzyme production:10g citrate,1.2g NaNO3,0.5gMgSO4, 0.5g K2HPO4·7H2O,0.005g CaCl2,and 0.0005g ZnSO4·7H2O per litre.The optimum cultural condition for enzyme production in shaking flask:temperature 30℃,initial pH 7.0,medium volume 50ml/150ml flask, inoculum concentration 2%,shaking speed 150r/min,culture period 72 hours.The optimum temperature and pH for enzyme production during the culture:30℃and 7.5,respectively.The enzyme was very sensitive to temperature.At the optimum temperature(30℃),the purified enzyme could be stored for 150min.Inactivation of the enzyme was very quick with the increase of temperature.When temperature was as high as 50℃,the enzymatic activity would completely lost within 5min.The enzyme was relative stable in the range of pH 7.0～8.0.basic solution was more harmful to enzymatic activity than acidic.The oxidizing activity of the enzyme could be activated by Ca2+,Mg2+ and Zn2+,whereas Cu2+,Hg2+ and Al3+ could inhibit the enzymatic activity.No effect of Fe 2+,K+ and Na+ on enzymatic activity was observed.
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Pollution Control,2002,3(9):35-37.[NB140晨兴,许旭萍,林跃鑫,谢华玲,李惠珍.鞘细菌FC9901氧化铁生化机制的研究.环境污染治理技术与设备,2002,3(9):35-37] [11[ De Vrind J,Jong E W, Corstjens P.Oxidation of manganese and iron by Leptothrix discophora:Use of TMPD as an indicator of metal oxidation[J].Appl.Environ.Microbiol.1990,56:3458-3462 [12[ Corstjens P, De Vrind J.Enzymatic iron oxidation by Leptothrix discophora:identification of Mn2+ oxidizing protein from Leptothrix discophora ss-1[J].Appl.Environ.Microbiol,1992,58:450-454 [13[ State Environmental Protection Administration.Analytical methods of water and water monitoring[M].Beijing:China Environmental Science Press,1989.[国家环保局主编,水和水监测分析方法,北京:中国环境科学出版社,1989] [14[ APHA AWWA&WPCF.Standardized examination methods of water and sewage(15th edition)[M].Chinese edition translated by Song R Y et al.Beijing:China Architecture Industry Press,1985.[宋仁元等译,APHA AWWA&WPCF,水和废水标准检验法(第十五版),北京:中国建筑工业出版社,1985]
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