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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 Fe 2+ OXIDASE PRODUCTION OF Fe 2+OXIDIZED SHEATHED BACTERIA SPHAEROTILUS NATANS FC9901 AND SOME ENZYMATIC CHARACTERISTICS OF Fe 2+OXIDASE
【摘要】 对氧化铁鞘细菌FC990 1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 ,NaNO3 1.2g,MgSO4·7H2 O 0 .5g ,K2 HPO4·7H2 O 0 .5g ,CaCl2 0 .0 15g ,ZnSO4·7H2 O 0 .0 0 0 5g。最适产酶条件为 :温度 30℃ ,起始pH7.0 ,接种量 2 % ,15 0mL三角瓶装 5 0mL ,15 0r/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+-Mn 2+oxidizing sheathed bacteria is a group of bacteria with ability to oxidize Fe 2+ and Mn 2+.In practice they were used to treat wastewater.Previous researches indicated that the functional component for Fe 2+-Mn 2+ oxidation in the bacterium is Fe 2+oxidase.Because of the low content and easy inactivation,Fe 2+oxidase is difficult to be purified.This resulted the less information about the properties of the enzyme.A strain of Fe 2+ oxidizing sheathed bacteria Sphaerotilus natans FC9901 we separated previous was used as material for Fe 2+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 NaNO 3,0.5gMgSO 4, 0.5g K 2HPO 4·7H 2O,0.005g CaCl 2,and 0.0005g ZnSO 4·7H 2O 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 Ca 2+,Mg 2+ and Zn 2+,whereas Cu 2+,Hg 2+ and Al 3+ could inhibit the enzymatic activity.No effect of Fe 2+,K+ and Na+ on enzymatic activity was observed.
【Key words】 Oxidized sheathed bacteria; Fe 2+ oxidase; Enzyme-producing conditions; Enzymatic characteristics.;
- 【文献出处】 水生生物学报 ,Acta Hydrobiologica Sinica , 编辑部邮箱 ,2004年04期
- 【分类号】Q936
- 【被引频次】11
- 【下载频次】147