Aerobic dechlorination of cis- and trans-dichloroethenes by some indigenous bacteria isolated from contaminated sites in Africa

IntroductionChlorinatedethenesareamongthemostcommoncontaminantsobservedingroundwatersystems (Bouwer,1994;McCarty,1994) .Thesecompoundsvaryinnumberofchlorinesubstituents,fromthemostchlorinated,terachloroethene (PCE ) ,tothemonochlorinatedvinylchloride (VC …     

Individual stages of bacterial dichloromethane degradation mapped by carbon and chlorine stable isotope analysis

The fractionation of carbon and chlorine stable isotopes of dichloromethane(CH_2Cl_2,DCM)upon dechlorination by cells of the aerobic methylotroph Methylobacterium extorquens DM4 and by purified DCM dehalogenases of the glutathione S-transferase family was analyzed.Isotope effects for individual steps of the multi-stage DCM degradation process,including transfer across the cell wall from the aqueous medium to the cell cytoplasm,dehalogenase binding,and catalytic reaction,were considered.The observed carbon and chlorine isotope fractionation accompanying DCM consumption by cell supensions and enzymes was mainly determined by the breaking of C\Cl bonds,and not by inflow of DCM into cells.Chlorine isotope effects of DCM dehalogenation were initially masked in high density cultures,presumably due to inverse isotope effects of non-specific DCM oxidation under conditions of oxygen excess.Glutathione cofactor supply remarkably affected the correlation of variations of DCM carbon and chlorine stable isotopes(Δδ~(13)C/Δδ~(37)Cl),increasing corresponding ratio from 7.2–8.6 to 9.6–10.5 under conditions of glutathione deficiency.This suggests that enzymatic reaction of DCM with glutathione thiolate may involve stepwise breaking and making of bonds with the carbon atom of DCM,unlike the uncatalyzed reaction,which is a one-stage process,as shown by quantum-chemical modeling.     

细菌二氯甲烷脱氯酶基因对拟南芥的转化

卤代烷烃是一类很普遍的环境污染物 ,在工业生产中常伴随着废气废物释放出来 ,严重污染土壤及地下水 ,并且严重破坏臭氧层 .在此类污染物含量较高的土壤中分离出一些细菌 ,它们能以卤代烷烃为唯一碳源进行生长 ,且将这些有毒废物分解成无毒的化合物 .我们曾经将细菌卤代烷烃脱卤酶基因克隆到拟南芥中并得到高效表达[1] .Ｍｅｔｈｙｌｏｂａｃｔｅｒｉｕｍ和Ｈｙ ｐｈｏｍｉｃｒｏｂｉｕｍ属的细菌能利用二氯甲烷作为唯一碳源进行生长[5] ,其代谢过程如图 1.其中参与反应的关键酶是二氯甲烷脱氯酶 ,此酶属于谷胱甘肽转移酶类 ,在脱ＣＩ时 ,先…     

二噁英降解酶的研究进展

二噁英是一类广泛分布于环境中的持久性有机污染物,它们能够在食物链中逐级积累,并可引发多种细胞毒性,严重影响生态系统和人类健康。在众多二噁英降解方法中,生物降解法以环境友好和低成本等优点一直受到国内外学者的青睐,而二噁英降解酶是生物降解法中的关键物质。简要综述了近年来二噁英典型降解酶的降解机制、效果和编码基因等方面研究进展,并对二噁英降解酶未来的研究和应用方向进行了展望。     

Chloromuconolactone dehalogenase ClcF of actinobacteria

This work investigated the distribution of the clcF gene in actinobacteria isolated from different ecotopes. The gene encodes chloromuconolactone dehalogenase (CMLD) ClcF, the enzyme found to date in only one representative of Gram‐positive bacteria, Rhodococcus opacus 1CP, adapted to 2‐chlorophenol (2CP). Using primers specific to the clcF gene, from the DNA matrix of rhodococcal strains closely related to species Rhodococcus wratislaviensis (P1, P12, P13, P20, G10, KT112, KT723, BO1) we obtained PCR products whose nucleotide sequences were 100% identical to that of the clcF gene from strain R. opacus 1CP. CMLDs isolated from the biomass of strains Rhodococcus spp. G10 and P1 grown on 2CP did not differ by their subunit molecular mass deduced from the known amino acid sequence of the clcF gene from the ClcF of strain R. opacus 1CP. Matrix‐assisted laser dissociation/ionization time‐of‐flight mass spectrometry showed the presence of a peak with m/z 11,194–11,196 Da both in whole cells and in protein solutions with a ClcF activity. Thus, we have first time shown the distribution of ClcF among actinobacteria isolated from geographically distant habitats.     

假单胞菌No.66降解氯乙酸的研究

本文介绍从水稻土分离得一株可利用单氯乙酸和二氯乙酸作唯一碳源的假单胞菌No.66。该菌在二氯乙酸或单氯乙酸基质上生长时,会释放氯离子,并能使二氯乙酸为碳源的基质之pH值由7降至5以下。No.66菌适宜生长的氯乙酸浓度为0.4—0.5％,适宜脱氯的pH为6.0。所产生的脱氯酶与碳源种类有关。粗脱氯酶液在4℃进行聚丙烯酰胺凝胶(10.5％)电泳呈现一条酶活力带,该酶不仅能使单氯乙酸和二氯乙酸脱氯,而且也能使三氯乙酸脱氯。