大豆卵磷脂和乐果有机磷降解菌的分离鉴定及酶活性比较
发布时间：2018-11-24 13:21
【摘要】：【目的】明确2株有机磷降解菌Yj2和Yj3对大豆卵磷脂和乐果有机磷的降解特性及酶活性。【方法】利用16S r DNA鉴定从大豆土壤中分离得到的Yj2和Yj3菌株,在菌株最佳生长条件下,测定了不同磷源时菌体的酶活性并分级纯化了有机磷降解酶。【结果】Yj2为醋酸钙不动杆菌Acinetobacter sp.,其最佳生长碳源为葡萄糖,氮源为硫酸铵,p H为8;Yj3为芽孢杆菌Bacillus sp.,其最佳生长碳源为葡萄糖,氮源为蛋白胨,p H为9。大豆卵磷脂为磷源时,Yj3的菌体生长情况稍优于Yj2。乐果为磷源时,Yj2的菌体生长情况稍优于Yj3;72 h内Yj2酸性和碱性磷酸酶活性整体高于Yj3,而有机磷降解酶活性低于Yj3。硫酸铵沉淀法+阳离子交换层析分别从Yj2和Yj3菌体中成功分离纯化了有机磷降解酶,SDS-PAGE结果显示纯化的蛋白均为单一条带。Yj2硫酸铵沉淀法+阳离子交换层析的提纯倍数是硫酸铵沉淀的7.77倍,硫酸铵沉淀为粗酶的1.35倍。Yj3硫酸铵沉淀法+阳离子交换层析的提纯倍数是硫酸铵沉淀的5.07倍,硫酸铵沉淀为粗酶的1.53倍。【结论】菌株Acinetobacter sp.Yj2和Bacillus sp.Yj3都具有降解大豆卵磷脂及乐果有机磷的特性,对有机磷降解起主要作用的是酸性磷酸酶、碱性磷酸酶及有机磷降解酶,它们在2株菌株对大豆卵磷脂和乐果降解过程中所起的作用有明显差异。可从Yj2和Yj3菌体分离纯化获得提纯倍数较高的有机磷降解酶蛋白。
[Abstract]:[objective] to identify the degradation characteristics and enzyme activities of soybean lecithin and dimethoate organophosphorus by two organophosphorus degrading bacteria (Yj2 and Yj3). [methods] the strains of Yj2 and Yj3 isolated from soybean soil were identified by 16s r DNA. Under the optimum growth conditions, the enzyme activity of the strain was determined and the organophosphorus degrading enzyme was purified by grade. [results] Yj2 was Acinetobacter cereus Acinetobacter sp.,. The best growth carbon source was glucose, and the nitrogen source was ammonium sulfate. P H = 8; The optimum growth carbon source was glucose and the nitrogen source was peptone, p H 9. When soybean lecithin was used as phosphorus source, the growth of Yj3 was slightly better than that of Yj2.. When dimethoate was a phosphorus source, the growth of Yj2 was slightly better than that of Yj2 acid and alkaline phosphatase activity in Yj3;72 h, while the activity of organophosphorus degrading enzyme was lower than that of Yj3.. Organophosphorus degrading enzymes were successfully isolated and purified from Yj2 and Yj3 by cation exchange chromatography with ammonium sulfate precipitation. The results of SDS-PAGE showed that the purified proteins were all single bands. The purification times of Yj2 ammonium sulfate precipitation chromatography were 7.77 times than those of ammonium sulfate precipitation. Ammonium sulfate precipitation was 1.35 times as high as that of crude enzyme. The purification ratio of Yj3 ammonium sulfate precipitation method was 5.07 times higher than that of ammonium sulfate precipitation. [conclusion] both Acinetobacter sp.Yj2 and Bacillus sp.Yj3 have the properties of degrading soybean lecithin and dimethoate organophosphorus, and acid phosphatase plays a major role in the degradation of organic phosphorus. Alkaline phosphatase (ALP) and organophosphorus degrading enzyme (OP-degrading enzyme) played different roles in the degradation of soybean lecithin and dimethoate by two strains. High purification times of organophosphorus degrading enzyme protein were obtained from Yj2 and Yj3.
【作者单位】： 吉林农业大学生命科学学院;吉林农业大学农学院;
【基金】：国家自然科学基金(31201687)
【分类号】：X592;X172
本文编号：2353883
[Abstract]:[objective] to identify the degradation characteristics and enzyme activities of soybean lecithin and dimethoate organophosphorus by two organophosphorus degrading bacteria (Yj2 and Yj3). [methods] the strains of Yj2 and Yj3 isolated from soybean soil were identified by 16s r DNA. Under the optimum growth conditions, the enzyme activity of the strain was determined and the organophosphorus degrading enzyme was purified by grade. [results] Yj2 was Acinetobacter cereus Acinetobacter sp.,. The best growth carbon source was glucose, and the nitrogen source was ammonium sulfate. P H = 8; The optimum growth carbon source was glucose and the nitrogen source was peptone, p H 9. When soybean lecithin was used as phosphorus source, the growth of Yj3 was slightly better than that of Yj2.. When dimethoate was a phosphorus source, the growth of Yj2 was slightly better than that of Yj2 acid and alkaline phosphatase activity in Yj3;72 h, while the activity of organophosphorus degrading enzyme was lower than that of Yj3.. Organophosphorus degrading enzymes were successfully isolated and purified from Yj2 and Yj3 by cation exchange chromatography with ammonium sulfate precipitation. The results of SDS-PAGE showed that the purified proteins were all single bands. The purification times of Yj2 ammonium sulfate precipitation chromatography were 7.77 times than those of ammonium sulfate precipitation. Ammonium sulfate precipitation was 1.35 times as high as that of crude enzyme. The purification ratio of Yj3 ammonium sulfate precipitation method was 5.07 times higher than that of ammonium sulfate precipitation. [conclusion] both Acinetobacter sp.Yj2 and Bacillus sp.Yj3 have the properties of degrading soybean lecithin and dimethoate organophosphorus, and acid phosphatase plays a major role in the degradation of organic phosphorus. Alkaline phosphatase (ALP) and organophosphorus degrading enzyme (OP-degrading enzyme) played different roles in the degradation of soybean lecithin and dimethoate by two strains. High purification times of organophosphorus degrading enzyme protein were obtained from Yj2 and Yj3.
【作者单位】： 吉林农业大学生命科学学院;吉林农业大学农学院;
【基金】：国家自然科学基金(31201687)
【分类号】：X592;X172
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