【摘要】：【目的】明確2株有機(jī)磷降解菌Yj2和Yj3對(duì)大豆卵磷脂和樂果有機(jī)磷的降解特性及酶活性�！痉椒ā坷�16S r DNA鑒定從大豆土壤中分離得到的Yj2和Yj3菌株,在菌株最佳生長條件下,測(cè)定了不同磷源時(shí)菌體的酶活性并分級(jí)純化了有機(jī)磷降解酶�！窘Y(jié)果】Yj2為醋酸鈣不動(dòng)桿菌Acinetobacter sp.,其最佳生長碳源為葡萄糖,氮源為硫酸銨,p H為8;Yj3為芽孢桿菌Bacillus sp.,其最佳生長碳源為葡萄糖,氮源為蛋白胨,p H為9。大豆卵磷脂為磷源時(shí),Yj3的菌體生長情況稍優(yōu)于Yj2。樂果為磷源時(shí),Yj2的菌體生長情況稍優(yōu)于Yj3;72 h內(nèi)Yj2酸性和堿性磷酸酶活性整體高于Yj3,而有機(jī)磷降解酶活性低于Yj3。硫酸銨沉淀法+陽離子交換層析分別從Yj2和Yj3菌體中成功分離純化了有機(jī)磷降解酶,SDS-PAGE結(jié)果顯示純化的蛋白均為單一條帶。Yj2硫酸銨沉淀法+陽離子交換層析的提純倍數(shù)是硫酸銨沉淀的7.77倍,硫酸銨沉淀為粗酶的1.35倍。Yj3硫酸銨沉淀法+陽離子交換層析的提純倍數(shù)是硫酸銨沉淀的5.07倍,硫酸銨沉淀為粗酶的1.53倍�！窘Y(jié)論】菌株Acinetobacter sp.Yj2和Bacillus sp.Yj3都具有降解大豆卵磷脂及樂果有機(jī)磷的特性,對(duì)有機(jī)磷降解起主要作用的是酸性磷酸酶、堿性磷酸酶及有機(jī)磷降解酶,它們?cè)?株菌株對(duì)大豆卵磷脂和樂果降解過程中所起的作用有明顯差異�？蓮腨j2和Yj3菌體分離純化獲得提純倍數(shù)較高的有機(jī)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.
【作者單位】： 吉林農(nóng)業(yè)大學(xué)生命科學(xué)學(xué)院;吉林農(nóng)業(yè)大學(xué)農(nóng)學(xué)院;
【基金】：國家自然科學(xué)基金(31201687)
【分類號(hào)】：X592;X172

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