Fortin N, Fulthorpe R R, Allen D G, Greer C W
Biotechnology Research Institute, National Research Council of Canada, Montréal, QC, Canada.
Can J Microbiol. 1998 Jun;44(6):537-46.
Chloroaliphatics are major components of bleached kraft mill effluents. Gene probes and oligonucleotide primers were developed to monitor kraft pulp mill effluent treatment systems for the presence of key genes (dehalogenases) responsible for the dehalogenation of chloroaliphatic organics. The primers were used for polymerase chain reaction (PCR) analysis of genomic DNA extracted from dehalogenating bacterial isolates and from total community DNA extracted from water and sediments of mill effluent treatment system. PCR amplification with oligonucleotide primers designed from dhlB, encoding the haloacid dehalogenase from Xanthobacter autotrophicus, revealed the presence of dehalogenase genes in both aerated lagoons and stabilization basins. Similarly, positive results were obtained with mmoX primers designed from the soluble methane monooxygenase gene of Methylococcus capsulatus Bath. The haloacetate dehalogenase encoding gene (dehH2) from Moraxella sp. was typically not detected in mill effluent treatment systems unless the biomass was selectively enriched. DNA sequence analysis of several PCR fragaments revealed significant similarity to known dehalogenase amd methane monooxygenase genes. The results indicated a broad distribution of known dehalogenation genes and bacteria with chloroorganic-degrading potential in the mill effluent treatment systems.
氯代脂肪族化合物是漂白硫酸盐制浆厂废水的主要成分。开发了基因探针和寡核苷酸引物，用于监测硫酸盐制浆厂废水处理系统中负责氯代脂肪族有机物脱卤作用的关键基因（脱卤酶）的存在情况。这些引物用于对从脱卤细菌分离株中提取的基因组DNA以及从制浆厂废水处理系统的水和沉积物中提取的总群落DNA进行聚合酶链反应（PCR）分析。用根据编码自养黄色杆菌卤代酸脱卤酶的dhlB设计的寡核苷酸引物进行PCR扩增，结果表明曝气塘和稳定塘中均存在脱卤酶基因。同样，用根据荚膜甲基球菌可溶性甲烷单加氧酶基因设计的mmoX引物也得到了阳性结果。除非生物量被选择性富集，否则在制浆厂废水处理系统中通常检测不到来自莫拉克斯氏菌属的卤乙酸脱卤酶编码基因（dehH2）。对几个PCR片段的DNA序列分析表明，它们与已知的脱卤酶和甲烷单加氧酶基因具有显著的相似性。结果表明，制浆厂废水处理系统中已知的脱卤基因和具有氯代有机物降解潜力的细菌分布广泛。
