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对通过在萘和联苯上生长而分离出的菌株进行基因与双加氧酶探针杂交及多氯联苯降解能力的评估。

Evaluation of strains isolated by growth on naphthalene and biphenyl for hybridization of genes to dioxygenase probes and polychlorinated biphenyl-degrading ability.

作者信息

Pellizari V H, Bezborodnikov S, Quensen J F, Tiedje J M

机构信息

Center for Microbial Ecology, Michigan State University, East Lansing, 48824, USA.

出版信息

Appl Environ Microbiol. 1996 Jun;62(6):2053-8. doi: 10.1128/aem.62.6.2053-2058.1996.

Abstract

Approximately equal numbers of bacteria were isolated from primarily tropical soils by growth on biphenyl and naphthalene to compare their competence in polychlorinated biphenyl (PCB) degradation. The strains isolated by growth on biphenyl catalyzed more extensive PCB degradation than the strains isolated by growth on naphthalene, suggesting that naphthalene cocontamination may be only partially effective in stimulating the cometabolism of lower chlorinated PCBs. Probes were made from the bph, nah, and tod genes encoding the large iron iron sulfur protein of the dioxygenase complex and hybridized to 19 different strains. The hybridization patterns did not correlate well with the substrates of isolation, suggesting that there is considerable diversity in these genes in nature and that probe hybridization is not a reliable indication of catabolic capacity. The strains with the most extensive PCB degradation capacity did strongly hybridize to the bph probe, but a few strains that exhibited strong hybridization had poor PCB-degrading ability. Of the 19 strains studied, 5 hybridized to more than one probe and 2, including one strong PCB degrader, hybridized to all three probes. Southern blots showed that the bph and nah probes hybridized to separate bands, suggesting that multiple dioxygenases were present. Multiple dioxygenases may be an important feature of competitive decomposers in nature and hence may not be rare. Most of the isolates identified were members of the beta subgroup of the Proteobacteria, a few were gram positive, and none were true Pseudomonas species.

摘要

通过在联苯和萘上生长,从主要是热带土壤中分离出数量大致相等的细菌,以比较它们在多氯联苯(PCB)降解方面的能力。通过在联苯上生长分离出的菌株比通过在萘上生长分离出的菌株催化更广泛的PCB降解,这表明萘共污染在刺激低氯代PCB的共代谢方面可能仅部分有效。从编码双加氧酶复合物的大铁铁硫蛋白的bph、nah和tod基因制备探针,并与19种不同的菌株杂交。杂交模式与分离的底物相关性不佳,这表明这些基因在自然界中存在相当大的多样性,并且探针杂交不是分解代谢能力的可靠指标。具有最广泛PCB降解能力的菌株确实与bph探针强烈杂交,但一些表现出强杂交的菌株PCB降解能力较差。在所研究的19个菌株中,5个与不止一个探针杂交,2个包括一个强PCB降解菌与所有三个探针杂交。Southern印迹显示bph和nah探针与不同的条带杂交,表明存在多种双加氧酶。多种双加氧酶可能是自然界中竞争性分解者的一个重要特征,因此可能并不罕见。鉴定出的大多数分离物是变形菌门β亚群的成员,少数是革兰氏阳性菌,没有一个是真正的假单胞菌属物种。

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