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

【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,...

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Published in华南农业大学学报 Vol. 37; no. 2; pp. 65 - 72
Main Author 杨美英 于亭 王春红 孙合美 刘晶晶 武志海
Format Journal Article
LanguageChinese
Published 吉林农业大学生命科学学院,吉林长春,130118%吉林农业大学农学院,吉林长春,130118 2016
Subjects
乐果
大豆卵磷脂
有机磷降解酶
芽孢杆菌
醋酸钙不动杆菌
dimethoate
Acinetobacter calcoaceticus
乐果
有机磷降解酶
soy lecithin
醋酸钙不动杆菌
Bacillus
芽孢杆菌
organophosphate degrading enzyme
大豆卵磷脂
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ISSN1001-411X
DOI10.7671/j.issn.1001-411X.2016.02.010

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Summary:【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。
Bibliography:Acinetobacter calcoaceticus ; Bacillus ; dimethoate ; soy lecithin ; organophosphate degrading enzyme
44-1110/S
[ Objective] In order to clarify degradation characteristics and ＇enzyme activities of two organic phosphorus degrading bacteria in soy lecithin and dimethoate. [ Method ] Bacterium strains Yj2 and Yj3 were isolated from soy soil and identified by 16S rDNA identification method. The growth conditions of Yj2 and Yj3 were optimized by orthogonal test. The enzyme activities from two strains were determined under different phosphorus sources, and organophosphate degrading enzymes were classified and purified. [ Result] Yj2 was identified as Acinetobacter sp. , and Yj3 was identified as Bacillus sp.. The result of growth condition optimization showed that the best carbon and nitrogen source and pH values for strain Yj2 and Yj3 were glucose, ammonium sulfate, pH 8 and glucose, peptone, pH 9, respectively. The growth of Yj3 was slightly better than that of Yj2 when the phosphorus source was soy lecithin. How
ISSN:1001-411X
DOI:10.7671/j.issn.1001-411X.2016.02.010