精噁唑禾草灵的微生物降解
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摘要
本研究取安徽华星化工有限公司精噁唑禾草灵废水处理系统进水口处污泥进行驯化培养，从中筛选出活性菌株，对其中降解精噁唑禾草灵效果最好的活性菌株进行了鉴定以及UV诱变，并以此菌株以及此菌株UV诱变菌株(诱变菌株中降解精噁唑禾草灵效果最好)为出发菌株，运用高效液相色谱分析精噁唑禾草灵残存量的方法，进行了降解动力学以及外源物质对降解菌活性影响的研究，并通过高效薄层色谱方法，对精噁唑禾草灵的降解产物进行了分离。主要研究结果如下：
1．取安徽华星化工有限公司精噁唑禾草灵废水处理系统进水口处污泥进行驯化培养，分离到8株有效菌株，通过高效液相色谱分析这8株菌株降解精噁唑禾草灵的残存量，确定其中降解精噁唑禾草灵最好的一株菌株为本实验菌株，通过传统的微生物鉴定方法，将其鉴定为产碱菌属(Alcaligenes sp)，标记为Alcaligenes sp.H。
2．以Alcaligenes sp.H为出发菌株，通过紫外诱变后，从中挑选降解精噁唑禾草灵效果最好的诱变株H8-1为本实验的第二株菌株，标记为Alcaligenes sp.H_(uv)。在精噁唑禾草灵浓度为5-100mg／L范围内，Alcaligenes sp.H_(uv)菌降解精噁唑禾草灵的效果优于Alcaligenes sp.H。
3．本实验中Alcaligenes sp.H和Alcaligenes sp.H_(uv)以及混合菌降解精噁唑禾草灵的降解动力学方程用一级动力学方程模拟，其相关系数r均大于0.90。
C_t=C_0e~(-kt)
4．Alcaligenes sp.H与Alcaligenes sp.H_(uv)不同混合比例(2.5：1，1：1，1：2.5)的混合菌降解效果均明显优于Alcaligenes sp.H单菌，其半衰期分别为37.66h、30.39h、23.90h均明显小于Alcaligenes sp.H单菌的半衰期64.17h，而当Alcaligenes sp.H_(uv)在混合菌体系中所占比例超过一半时，混合菌的降解效果优于Alcaligenes sp.H_(uv)单菌(半衰期为35.18h)，同时，随着Alcaligenes sp.H与Alcaligenes sp.H_(uv)混合比例的下降，即Alcaligenes sp.H_(uv)菌的比例增大，降解加快，这一点可以从半衰期的逐渐减小得到反映，说明Alcaligenes sp.H_(uv)菌在混合菌体系中占主导地位，也间接反映其降解效果优于Alcaligenes sp.H。
5．Alcaligenes sp.H菌量分别为OD_(330)=0.2，0.4，0.6时，精噁唑禾草灵的微生物降解半衰期分别为67.28h、35.0h、21.72h，其半衰期与菌量近似成反相关，降解速率常数与Alcaligenes sp.H菌量近似成正比。而Alcaligenes sp.H_(uv)菌量分别为OD=0.2，0.4，0.6时，精噁唑禾草灵的微生物降解半衰期为35.54h、21.32h、19.58h，虽然其降解速率与Alcaligenes sp.H_(uv)菌量呈正相关，但其反映的菌量浓度差并没有Alcaligenes sp.H大，可见对于Alcaligenes sp.H_(uv)来说并非所有的菌都参与了降解(相对OD_(330)=0.2)，说明对于一定浓度的精噁唑禾草灵来说，随着降解菌菌量的增加，农药的降解加快，降解速率与菌量基本成正相关，但同样存在一个最佳菌量问题，即当
菌量增加到一定程度后，再增加菌量，其降解速率增加趋势不再明显。
6．在精嗯哇禾草灵的浓度从 sing／L到 100mg／卜变化范围内，对于 Alcalfgenes Sp．H
来说，其半衰期逐渐增大，分别为47．47h、65．38h、68．6lh、86．63h、135．88h，对于
Alcaliggnes sp．Huv来说其半衰期分另为 18．24h、35．54h、46．51h、57．27h、72．19h；
半衰期同样逐渐增大。不同精嗯哇禾草灵浓度对降解动力学的影响表明：AICJ人卯们“
sp．H及 Alcallee，7es s H。v只对一定浓度的精噬哇禾草灵保持较强的降解作用，
A儿7rnes sn·H及 Alcaliernes sn Huv降解高浓度的精曙哩禾草灵的效率下降。
7．本实验所选取的 3种外源物质对Aleal＠nes Sp．H及Alcalwnes Sp．HW降解精嗯哇
禾草灵均产生了抑制作用。其中百菌清对 AlcaligeneS SP．HW及 Alcaligenes Sp丑与
Alcaligenes Sp．H＊混合菌门：1）降解精嗯哇禾草灵的抑制作用更为显著，而对于
AlcaligenesspH其相对抑制作用较小；当外源物质为十二烷基苯磺酸钠和 Cr＋6时，降解
曲线形状发生了明显的变化，尤其降解初期十＝烷基苯磺酸钠和Cr＋‘对降解菌的抑制作
用尤为明显，降解速度很慢，随着降解菌对十＝烷基苯磺酸钠和 Cr＋0的适应，降解速度
逐渐加快。
8．在展开剂为甲苯：H氯甲烷（V：V，7：3）条件下，精嗯哇禾草灵降解产物得到很
好的分离，通过比较精嗓哇禾草灵标样及精嗯陛禾草灵降解产物的展开距离即R。值，
除Rf叫．22（精嗯哇禾草灵标样峰）的峰外，精嗯哩禾草灵降解产物（降解菌为
Ale。lfgenes SP．H）在 Rr＝0．14、0．29、0．34、0．39、0．47处有 5个峰；精lg陛禾草灵
降解产物（降解菌为 Alcalfgunes sp．Hm）在 Rr＝0．11、0．39处有 2个峰，这与 CAMAG紫
外灯下观察的斑点数量相符。通过 HPLC－MS分析可以推测出：m）《三－门－氯J，
3－苯并嗯陛千－基氧）苯氧基」丙酸乙酯水解是降解菌 Alcallernes p丑与 Alcalfgenes
SP．Huv降解精嗯咆禾草灵的公共降解途径之一，其产物为m）六二－历－氯习，卜苯
并嗯哇－2－基氧）苯氧基」丙酸和乙醇。
Bacterial strains which could decompose fenoxaprop-p-ethyl were isolated from a batch culture with fenoxaprop-p-ethyl as carbon source and the sludge from waste water dealing system of Huaxing chemical engineering Co L.td . After determining the rest quantity of fenoxaprop-p-ethyl with HPLC , H was selected as dominant degradation bacterial strain for its best degradation ability to fenoxaprop-p-ethyl . H8-1 was also selected as another dominant degradation bacterial strain for its higher degradation ability to fenoxaprop-p-ethyl than the other strains after UV mutation . Fenoxaprop-p-ethyl degradation dynamics was studied with HPLC method , and degradation products of fenoxaprop-p-ethyl had been separated with HPTLC . The main study results are as follows :
1 .Eight bacterial strains which could decompose fenoxaprop-p-ethyl were isolated from a batch culture with fenoxaprop-p-ethyl as carbon source and the sludge from waste water dealing system of Huaxing chemical engineering Co L.td . After determining the rest quantity of fenoxaprop-p-ethyl with HPLC , H was selected as dominant degradation bacterial strain for its best degradation ability to fenoxaprop-p-ethyl .Based on the morphological appearances and physiological characteristics ,H was identified as Alcaligenes sp and named as Alcaligenes sp.H .
2.H8-1 was also selected as another dominant degradation bacterial strain for its higher degradation ability to fenoxaprop-p-ethyl than the other strains after UV induce . H8-1 was marked as Alcaligenes sp. Huv . With the concentration of fenoxaprop-p-ethyl varing from 5mg/L to 100mg/L , Alcaligenes sp. Huvhas better degradation ability to fenoxaprop-p-ethyl than Alcaligenes sp.H .
3. fenoxaprop-p-ethyl degradation dynamics was studied . Degradation kinetics follow the first order kinetics .
4. It proved that degradation ability of mixed bacterial by Alcaligenes sp.H and Alcaligenes sp. Huv to fenoxaprop-p-ethyl is superior to Alcaligenes sp.H , superior to Alcaligenes sp. Huv after the quantity of Alcaligenes sp. Huv surpasses half , and with the quantity increasing of Alcaligenes sp. Huv , degradation ability of mixed bacterial by and Alcaligenes sp. Huv to fenoxaprop-p-ethyl increases respectively comparing their half-time . Their half-times are 37.66h, 30.39h and 23.90h to mixed bacterial (Alcaligenes sp.H : Alcaligenes sp. Huv 2.5:1,1:1,1:2.5 ) ,64.17h to Alcaligenes sp.H and 35.18h to Alcaligenes sp. Huv.
5. Degradation rates of fenoxaprop-p-ethyl were positively correlative to biomass .On the
contrary , the fortified concentration is opposite the degradative rate . It can be seen from their half-time and their half-time are 67.28h, 35.0h and 21.72h to Alcaligenes sp.H , 35.54h ,21.32h and 19.58h to Alcaligenes sp. Huv correlative to their different biomass .Results showed that biomass has a appropriately fortified concentration .When surpassing this biomass , the degradative rate can not grow obviously .
6. With the concentration of fenoxaprop-p-ethyl varing from 5mg/L to 100mg/L , the half-time of Alcaligenes sp.H and Alcaligenes sp. Huv increased respectively .The different half-time of Alcaligenes sp.H and Alcaligenes sp. Huv with different concentration are 47.47h,65.38h,68.61h,86.63h,135.88h and 18.24h,35.54h,46.51h,57.27h,72.19h .Effects of different concentration of fenoxaprop-p-ethyl to degradation showed that Alcaligenes sp.H and Alcaligenes sp. Huv had better degradation in some a concentration and the degradation ability to fenoxaprop-p-ethyl of Alcaligenes sp.H and Alcaligenes sp. Huv was obviously decreased in high concentration.
7. Chlorothalonil, dodecylbenfenxaprop -p-ethylene sulfonic aid sodium salt and Cr+6can inhibite the degradation ability of Alcaligenes sp.H , Alcaligenes sp. Huv and the mixed bacterial by Alcaligenes sp.H and Alcaligenes sp. Huv to fenoxaprop-p-ethyl. The degradation ability of Alcaligenes sp.H to fenoxaprop-p-ethyl was severely inhibited by Chlorothalonil. In early period , Alcaligenes sp.H , Alcaligenes sp. an
1．取安徽华星化工有限公司精噁唑禾草灵废水处理系统进水口处污泥进行驯化培养，分离到8株有效菌株，通过高效液相色谱分析这8株菌株降解精噁唑禾草灵的残存量，确定其中降解精噁唑禾草灵最好的一株菌株为本实验菌株，通过传统的微生物鉴定方法，将其鉴定为产碱菌属(Alcaligenes sp)，标记为Alcaligenes sp.H。
2．以Alcaligenes sp.H为出发菌株，通过紫外诱变后，从中挑选降解精噁唑禾草灵效果最好的诱变株H8-1为本实验的第二株菌株，标记为Alcaligenes sp.H_(uv)。在精噁唑禾草灵浓度为5-100mg／L范围内，Alcaligenes sp.H_(uv)菌降解精噁唑禾草灵的效果优于Alcaligenes sp.H。
3．本实验中Alcaligenes sp.H和Alcaligenes sp.H_(uv)以及混合菌降解精噁唑禾草灵的降解动力学方程用一级动力学方程模拟，其相关系数r均大于0.90。
C_t=C_0e~(-kt)
4．Alcaligenes sp.H与Alcaligenes sp.H_(uv)不同混合比例(2.5：1，1：1，1：2.5)的混合菌降解效果均明显优于Alcaligenes sp.H单菌，其半衰期分别为37.66h、30.39h、23.90h均明显小于Alcaligenes sp.H单菌的半衰期64.17h，而当Alcaligenes sp.H_(uv)在混合菌体系中所占比例超过一半时，混合菌的降解效果优于Alcaligenes sp.H_(uv)单菌(半衰期为35.18h)，同时，随着Alcaligenes sp.H与Alcaligenes sp.H_(uv)混合比例的下降，即Alcaligenes sp.H_(uv)菌的比例增大，降解加快，这一点可以从半衰期的逐渐减小得到反映，说明Alcaligenes sp.H_(uv)菌在混合菌体系中占主导地位，也间接反映其降解效果优于Alcaligenes sp.H。
5．Alcaligenes sp.H菌量分别为OD_(330)=0.2，0.4，0.6时，精噁唑禾草灵的微生物降解半衰期分别为67.28h、35.0h、21.72h，其半衰期与菌量近似成反相关，降解速率常数与Alcaligenes sp.H菌量近似成正比。而Alcaligenes sp.H_(uv)菌量分别为OD=0.2，0.4，0.6时，精噁唑禾草灵的微生物降解半衰期为35.54h、21.32h、19.58h，虽然其降解速率与Alcaligenes sp.H_(uv)菌量呈正相关，但其反映的菌量浓度差并没有Alcaligenes sp.H大，可见对于Alcaligenes sp.H_(uv)来说并非所有的菌都参与了降解(相对OD_(330)=0.2)，说明对于一定浓度的精噁唑禾草灵来说，随着降解菌菌量的增加，农药的降解加快，降解速率与菌量基本成正相关，但同样存在一个最佳菌量问题，即当
菌量增加到一定程度后，再增加菌量，其降解速率增加趋势不再明显。
6．在精嗯哇禾草灵的浓度从 sing／L到 100mg／卜变化范围内，对于 Alcalfgenes Sp．H
来说，其半衰期逐渐增大，分别为47．47h、65．38h、68．6lh、86．63h、135．88h，对于
Alcaliggnes sp．Huv来说其半衰期分另为 18．24h、35．54h、46．51h、57．27h、72．19h；
半衰期同样逐渐增大。不同精嗯哇禾草灵浓度对降解动力学的影响表明：AICJ人卯们“
sp．H及 Alcallee，7es s H。v只对一定浓度的精噬哇禾草灵保持较强的降解作用，
A儿7rnes sn·H及 Alcaliernes sn Huv降解高浓度的精曙哩禾草灵的效率下降。
7．本实验所选取的 3种外源物质对Aleal＠nes Sp．H及Alcalwnes Sp．HW降解精嗯哇
禾草灵均产生了抑制作用。其中百菌清对 AlcaligeneS SP．HW及 Alcaligenes Sp丑与
Alcaligenes Sp．H＊混合菌门：1）降解精嗯哇禾草灵的抑制作用更为显著，而对于
AlcaligenesspH其相对抑制作用较小；当外源物质为十二烷基苯磺酸钠和 Cr＋6时，降解
曲线形状发生了明显的变化，尤其降解初期十＝烷基苯磺酸钠和Cr＋‘对降解菌的抑制作
用尤为明显，降解速度很慢，随着降解菌对十＝烷基苯磺酸钠和 Cr＋0的适应，降解速度
逐渐加快。
8．在展开剂为甲苯：H氯甲烷（V：V，7：3）条件下，精嗯哇禾草灵降解产物得到很
好的分离，通过比较精嗓哇禾草灵标样及精嗯陛禾草灵降解产物的展开距离即R。值，
除Rf叫．22（精嗯哇禾草灵标样峰）的峰外，精嗯哩禾草灵降解产物（降解菌为
Ale。lfgenes SP．H）在 Rr＝0．14、0．29、0．34、0．39、0．47处有 5个峰；精lg陛禾草灵
降解产物（降解菌为 Alcalfgunes sp．Hm）在 Rr＝0．11、0．39处有 2个峰，这与 CAMAG紫
外灯下观察的斑点数量相符。通过 HPLC－MS分析可以推测出：m）《三－门－氯J，
3－苯并嗯陛千－基氧）苯氧基」丙酸乙酯水解是降解菌 Alcallernes p丑与 Alcalfgenes
SP．Huv降解精嗯咆禾草灵的公共降解途径之一，其产物为m）六二－历－氯习，卜苯
并嗯哇－2－基氧）苯氧基」丙酸和乙醇。
Bacterial strains which could decompose fenoxaprop-p-ethyl were isolated from a batch culture with fenoxaprop-p-ethyl as carbon source and the sludge from waste water dealing system of Huaxing chemical engineering Co L.td . After determining the rest quantity of fenoxaprop-p-ethyl with HPLC , H was selected as dominant degradation bacterial strain for its best degradation ability to fenoxaprop-p-ethyl . H8-1 was also selected as another dominant degradation bacterial strain for its higher degradation ability to fenoxaprop-p-ethyl than the other strains after UV mutation . Fenoxaprop-p-ethyl degradation dynamics was studied with HPLC method , and degradation products of fenoxaprop-p-ethyl had been separated with HPTLC . The main study results are as follows :
1 .Eight bacterial strains which could decompose fenoxaprop-p-ethyl were isolated from a batch culture with fenoxaprop-p-ethyl as carbon source and the sludge from waste water dealing system of Huaxing chemical engineering Co L.td . After determining the rest quantity of fenoxaprop-p-ethyl with HPLC , H was selected as dominant degradation bacterial strain for its best degradation ability to fenoxaprop-p-ethyl .Based on the morphological appearances and physiological characteristics ,H was identified as Alcaligenes sp and named as Alcaligenes sp.H .
2.H8-1 was also selected as another dominant degradation bacterial strain for its higher degradation ability to fenoxaprop-p-ethyl than the other strains after UV induce . H8-1 was marked as Alcaligenes sp. Huv . With the concentration of fenoxaprop-p-ethyl varing from 5mg/L to 100mg/L , Alcaligenes sp. Huvhas better degradation ability to fenoxaprop-p-ethyl than Alcaligenes sp.H .
3. fenoxaprop-p-ethyl degradation dynamics was studied . Degradation kinetics follow the first order kinetics .
4. It proved that degradation ability of mixed bacterial by Alcaligenes sp.H and Alcaligenes sp. Huv to fenoxaprop-p-ethyl is superior to Alcaligenes sp.H , superior to Alcaligenes sp. Huv after the quantity of Alcaligenes sp. Huv surpasses half , and with the quantity increasing of Alcaligenes sp. Huv , degradation ability of mixed bacterial by and Alcaligenes sp. Huv to fenoxaprop-p-ethyl increases respectively comparing their half-time . Their half-times are 37.66h, 30.39h and 23.90h to mixed bacterial (Alcaligenes sp.H : Alcaligenes sp. Huv 2.5:1,1:1,1:2.5 ) ,64.17h to Alcaligenes sp.H and 35.18h to Alcaligenes sp. Huv.
5. Degradation rates of fenoxaprop-p-ethyl were positively correlative to biomass .On the
contrary , the fortified concentration is opposite the degradative rate . It can be seen from their half-time and their half-time are 67.28h, 35.0h and 21.72h to Alcaligenes sp.H , 35.54h ,21.32h and 19.58h to Alcaligenes sp. Huv correlative to their different biomass .Results showed that biomass has a appropriately fortified concentration .When surpassing this biomass , the degradative rate can not grow obviously .
6. With the concentration of fenoxaprop-p-ethyl varing from 5mg/L to 100mg/L , the half-time of Alcaligenes sp.H and Alcaligenes sp. Huv increased respectively .The different half-time of Alcaligenes sp.H and Alcaligenes sp. Huv with different concentration are 47.47h,65.38h,68.61h,86.63h,135.88h and 18.24h,35.54h,46.51h,57.27h,72.19h .Effects of different concentration of fenoxaprop-p-ethyl to degradation showed that Alcaligenes sp.H and Alcaligenes sp. Huv had better degradation in some a concentration and the degradation ability to fenoxaprop-p-ethyl of Alcaligenes sp.H and Alcaligenes sp. Huv was obviously decreased in high concentration.
7. Chlorothalonil, dodecylbenfenxaprop -p-ethylene sulfonic aid sodium salt and Cr+6can inhibite the degradation ability of Alcaligenes sp.H , Alcaligenes sp. Huv and the mixed bacterial by Alcaligenes sp.H and Alcaligenes sp. Huv to fenoxaprop-p-ethyl. The degradation ability of Alcaligenes sp.H to fenoxaprop-p-ethyl was severely inhibited by Chlorothalonil. In early period , Alcaligenes sp.H , Alcaligenes sp. an
引文
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10 Issa S. Degradation of atrazine and isoproturon in the unsaturated zone:a study from Southern England. Pesticide Science, 1999,55(5):539-545
11 马文漪，孙丽伟，薄海航．微生物降解速灭菊酯的遗传毒性效应．南京大学学报．自然科学版，1988,24(3)：555-561
12 虞云龙，盛国英，傅家谟等．杀灭菊酯的微生物降解及酶促降解．环境科学，1997,18(2)5-7
13 虞云龙．农药的微生物降解性及酶促降解．博士论文．1995
14 李淑彬．微生物降解有机磷农药(甲胺磷)的研究．硕士论文．1998
15 谢冰，徐亚同．农药废水处理工艺研究．上海环境科学，1996，15(10)：28-30
16 Pemberton, J .M.,Wynne, E .C.Genetic engineering and biological detoxification/degradation of insecticides,in insecticide Microbiology,ed. By R.Lal. 1984,P147-168
17.Guenzi W.D.Pesticides in soil and Water. Soil sci. Soc. Amer. inc.Publisher, Madison,Wiscosin,1974,P133-151
18 Mandelbaum R.T. et al,Mineralization of the s-Triazing ofatrazine by stable bacterial mixed culture.Appl.Environ. Microbiol., 1993(59),1695-1701
19．(日)山本出，深见顺一主编，农药的代谢降解与毒理，北京：化工出版社，1991，P155-160
20.Behki, R.M.,Khan,Shahamat.Degradation of atrizin,prpazine ,and simazine by Rhodococcus strain B-30.J. Agric. Food chem.. 1994(42) 1237-1241
21 (日)土壤微生物研究会编，土壤微生物实验法，北京：科学出版社，1981P47-61，
22.Guth,J.A. Experimental approaches to studying the fate of pesticide in soil,Progress in Pesticide Biochemistry ,Vol.1,ed.by Huston. D.H. and .Roberts. T. R1981.P85-110
23.Logan B E. Adsorption and removal of pentachlorophenoo by white rot fungi in bactch culture .Water Res., 1994,128(7): 1533～1539
24 Katayama-Hirayama K et al. Biodegradation of phenol and monochlorophenols by yeast. Rhodotorula glutinis Water Sci. Technol.,1994, 30(9): 59～66
25 刘志培．贾省芬．单甲脒降解菌的分离筛选．微生物学通报，1995，22(5)；285-288
26 Hill,l.R,et al.Pesticide Microbiology. Academic Press,London. 1978
27 Bollag,J.M. Microbial transformation of pesticides,Adv. Appl.Microbiol, 1974, 18:75-79
28 Barik S. Metabolism of insecticides by microorganism,in insecticides Microbiology, ed.by Lal. R, 1984,P87-130
29 周乐民．浅谈有机杀虫剂的微生物降解．广西植保，1999,12(1)：33-36
30 崔中利，李顺鹏．化学农药的微生物降解及其机制．江苏环境科技，1998(3)：1-3
31 周群英．环境工程微生物学(第二版)．北京；高等教育出版社．2002，P72-83
32 Derbyshire,M.K. et al. Purification and characterization of an N-methylcarbamate pesticide hydrolyzing enzyme .J.Agric .Food. Chem, 1987(35):871-877.
33 Mileski,G.J.et al. Biodegradation of pentochlorophenoi by the white -rot fungus Phanerochaete chrysosporiun.Appl.Environ. Microbiol, 1998(54):2885-2889.
34 Munnecke,D.M.. Enzymaticdetoxificationof waste organophosphate pesfcide.J.Agric. Food Chem.,1980(28):105-111.
35 Marty, J.L., Vouges. J. Purification and properties of a phenylcarbamate herbicide degrading enzyme of pseudomonas alcaligenes isolated from soil Agric. Biol.Chem., 1987(51):3287-3294
36 Sarkar. J.M.,et al.Immobilization of enzymes on clays and soils.Soil Biol. Biochem.,1989(21):223-230
37 Ruggiero,P. et al..Detoxification of 2,4-dichlorophenol by a laccase immobilized on soil orclay. Soil Sci, 1989(147):361-370
38 Mulbury.W.W, Karns.J.S.Parathion hydrolase specified by the Flavobacterium opd genes:relationship between the gene and protein .J.Bacteriol. 1989(171):6740-6746
39 Serdar .C M.,Gibson. D.T. Enzymatic hydrolysis of organophosphates: cloning and expression of a parathion hydrolase from Pseudomonas diminuta .Bio/Technology, 1985(7): 1151-1155
40 Don,R.H., Pemberton. J M.. Properities of six pesticide degradation plasmids isolated from Alcaligenes eutrophus and Alcaligenes paradoxus .J.Bacteriol, 1981(145):681-686
41 Amy, P.S.,et al. Characterization of aquatic bacterial and cloning of genes specifying partial degradation of 2,4-dichlorophenoxyacetic acid .Appl.Environ. Microbiol, 1983(49): 1237-1245
42 Don,R.H., Pemberton. J.M. Genetic and physical map of the 2,4-dichlorophenoxyacetic.acid. degradative.plasmid.pJP4.J.Bacterol, 1985(161):466-468
43 Don,R.H.,et al. Transposon mutagenesis and cloning analysis of the payhway for degradation of 2,4-dichlorophenoxyacetic acid and 3-chlorobenzoate in Alcaligenes eutrophus JMP 134(pJP4).J.Bacteriol, 1985(161):85-90
44 Kukor,J.,et al .Recruitment of a chromosomally encoded malelactate reductase for degradation of 2,4-dichlorophenoxyacetic acid by the plasmid pJP4.J.Bacteriol, 1989(171):3385-3390
45 Don R H, Pemberton J M.Genetic and physical map of the 2,4-dihlorophenoxyacetic acid degradative plasmid pJP4.J.Bacteriol. 161:466-468
46 Mulbry ,W.W., Keamey, P.C. Degradation of pesticides by microorganism and the potential for genetic manipulation ,Crop Protection,1991,10:334-346
47 闰艳春，姚良同，宋晓妍等．工程菌及其固定化细胞对有机磷农药的降解．中国环境科学，2001，21(5)：411—416
48 Head I M, Saunders J R,Pickup R W. Microbial evolution ,diversity, and ecology :A decade of ribosomal RNA analysis of uncultivated microorganisms. Microbiol Ecol, 1998.35(1):1-21
49 蹇文英，东秀珠．定向进化同源基因在细茵系统进化研究中的应用．微生物通报，2000，27(5) 377—383
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53 蒋展鹏，杨宏伟．有机物生物降解性的鉴定方法及定量结构活性关系的研究．第一届全国环境化学学术研讨会论文集，2002,10．P346-347
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6．唐受印主编，废水处理工程．北京：化学工业出版社出版发行，1998年第1版
7 王学东．咪唑烟酸在非耕地环境中降解与代谢得研究．博士论文．2003
8．王保军，刘志培，杨惠芳．单甲脒农药得微生物降解代谢研究．环境科学学报，1998，18(3)：296-302
9.Daniel R,Michael A. A model describing pesticide bioavalability and biodegradation in soil,Soil Science Society of America Journal. 1997,(4): 1078-1084
10 Issa S. Degradation of atrazine and isoproturon in the unsaturated zone:a study from Southern England. Pesticide Science, 1999,55(5):539-545
11 马文漪，孙丽伟，薄海航．微生物降解速灭菊酯的遗传毒性效应．南京大学学报．自然科学版，1988,24(3)：555-561
12 虞云龙，盛国英，傅家谟等．杀灭菊酯的微生物降解及酶促降解．环境科学，1997,18(2)5-7
13 虞云龙．农药的微生物降解性及酶促降解．博士论文．1995
14 李淑彬．微生物降解有机磷农药(甲胺磷)的研究．硕士论文．1998
15 谢冰，徐亚同．农药废水处理工艺研究．上海环境科学，1996，15(10)：28-30
16 Pemberton, J .M.,Wynne, E .C.Genetic engineering and biological detoxification/degradation of insecticides,in insecticide Microbiology,ed. By R.Lal. 1984,P147-168
17.Guenzi W.D.Pesticides in soil and Water. Soil sci. Soc. Amer. inc.Publisher, Madison,Wiscosin,1974,P133-151
18 Mandelbaum R.T. et al,Mineralization of the s-Triazing ofatrazine by stable bacterial mixed culture.Appl.Environ. Microbiol., 1993(59),1695-1701
19．(日)山本出，深见顺一主编，农药的代谢降解与毒理，北京：化工出版社，1991，P155-160
20.Behki, R.M.,Khan,Shahamat.Degradation of atrizin,prpazine ,and simazine by Rhodococcus strain B-30.J. Agric. Food chem.. 1994(42) 1237-1241
21 (日)土壤微生物研究会编，土壤微生物实验法，北京：科学出版社，1981P47-61，
22.Guth,J.A. Experimental approaches to studying the fate of pesticide in soil,Progress in Pesticide Biochemistry ,Vol.1,ed.by Huston. D.H. and .Roberts. T. R1981.P85-110
23.Logan B E. Adsorption and removal of pentachlorophenoo by white rot fungi in bactch culture .Water Res., 1994,128(7): 1533～1539
24 Katayama-Hirayama K et al. Biodegradation of phenol and monochlorophenols by yeast. Rhodotorula glutinis Water Sci. Technol.,1994, 30(9): 59～66
25 刘志培．贾省芬．单甲脒降解菌的分离筛选．微生物学通报，1995，22(5)；285-288
26 Hill,l.R,et al.Pesticide Microbiology. Academic Press,London. 1978
27 Bollag,J.M. Microbial transformation of pesticides,Adv. Appl.Microbiol, 1974, 18:75-79
28 Barik S. Metabolism of insecticides by microorganism,in insecticides Microbiology, ed.by Lal. R, 1984,P87-130
29 周乐民．浅谈有机杀虫剂的微生物降解．广西植保，1999,12(1)：33-36
30 崔中利，李顺鹏．化学农药的微生物降解及其机制．江苏环境科技，1998(3)：1-3
31 周群英．环境工程微生物学(第二版)．北京；高等教育出版社．2002，P72-83
32 Derbyshire,M.K. et al. Purification and characterization of an N-methylcarbamate pesticide hydrolyzing enzyme .J.Agric .Food. Chem, 1987(35):871-877.
33 Mileski,G.J.et al. Biodegradation of pentochlorophenoi by the white -rot fungus Phanerochaete chrysosporiun.Appl.Environ. Microbiol, 1998(54):2885-2889.
34 Munnecke,D.M.. Enzymaticdetoxificationof waste organophosphate pesfcide.J.Agric. Food Chem.,1980(28):105-111.
35 Marty, J.L., Vouges. J. Purification and properties of a phenylcarbamate herbicide degrading enzyme of pseudomonas alcaligenes isolated from soil Agric. Biol.Chem., 1987(51):3287-3294
36 Sarkar. J.M.,et al.Immobilization of enzymes on clays and soils.Soil Biol. Biochem.,1989(21):223-230
37 Ruggiero,P. et al..Detoxification of 2,4-dichlorophenol by a laccase immobilized on soil orclay. Soil Sci, 1989(147):361-370
38 Mulbury.W.W, Karns.J.S.Parathion hydrolase specified by the Flavobacterium opd genes:relationship between the gene and protein .J.Bacteriol. 1989(171):6740-6746
39 Serdar .C M.,Gibson. D.T. Enzymatic hydrolysis of organophosphates: cloning and expression of a parathion hydrolase from Pseudomonas diminuta .Bio/Technology, 1985(7): 1151-1155
40 Don,R.H., Pemberton. J M.. Properities of six pesticide degradation plasmids isolated from Alcaligenes eutrophus and Alcaligenes paradoxus .J.Bacteriol, 1981(145):681-686
41 Amy, P.S.,et al. Characterization of aquatic bacterial and cloning of genes specifying partial degradation of 2,4-dichlorophenoxyacetic acid .Appl.Environ. Microbiol, 1983(49): 1237-1245
42 Don,R.H., Pemberton. J.M. Genetic and physical map of the 2,4-dichlorophenoxyacetic.acid. degradative.plasmid.pJP4.J.Bacterol, 1985(161):466-468
43 Don,R.H.,et al. Transposon mutagenesis and cloning analysis of the payhway for degradation of 2,4-dichlorophenoxyacetic acid and 3-chlorobenzoate in Alcaligenes eutrophus JMP 134(pJP4).J.Bacteriol, 1985(161):85-90
44 Kukor,J.,et al .Recruitment of a chromosomally encoded malelactate reductase for degradation of 2,4-dichlorophenoxyacetic acid by the plasmid pJP4.J.Bacteriol, 1989(171):3385-3390
45 Don R H, Pemberton J M.Genetic and physical map of the 2,4-dihlorophenoxyacetic acid degradative plasmid pJP4.J.Bacteriol. 161:466-468
46 Mulbry ,W.W., Keamey, P.C. Degradation of pesticides by microorganism and the potential for genetic manipulation ,Crop Protection,1991,10:334-346
47 闰艳春，姚良同，宋晓妍等．工程菌及其固定化细胞对有机磷农药的降解．中国环境科学，2001，21(5)：411—416
48 Head I M, Saunders J R,Pickup R W. Microbial evolution ,diversity, and ecology :A decade of ribosomal RNA analysis of uncultivated microorganisms. Microbiol Ecol, 1998.35(1):1-21
49 蹇文英，东秀珠．定向进化同源基因在细茵系统进化研究中的应用．微生物通报，2000，27(5) 377—383
50 王连生，韩朔映．有机物定量结构—活性相关[M]．北京：中国环境科学出版社，1993
51 白遒彬．定量构效关系在环境化学中的应用．环境科学.,1990,11(5)：66-70
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