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Limited aromatic pathway genes diversity amongst aromatic compound degrading soil bacterial isolates

机译:芳香化合物降解土壤细菌分离物中有限的芳香途径基因多样性

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Identification and characterization of novel genes belonging to microbial aromatic biodegradation pathway is of great importance as they have been proven versatile biocatalysts. In this study, the selection of 19 environmental bacterial isolates capable to degrade a wide range of aromatic compounds has been screened for the presence of five genes from the lower and the upper aromatic biodegradation pathway using PCR methodology. In the case of 4-oxalocrotonate tautomerase and toluene dioxygenases, although present in the most of environmental isolates, very limited diversity of the genes has been encountered. Highly conserved sequences of these genes in environmental samples revealed high homology with gene sequences of the characterized corresponding genes from Pseudomonas putida species. The screen using degenerate primers based on known catechol-and naphthalene dioxygenases sequences resulted in a limited number of amplified fragments. Only two catechol 2,3-dioxygenase from two Bacillus isolates were amplified and showed no significant similarities with dioxygenases from characterized organisms, but 80-90% identities with partial catechol 2,3-dioxygenase sequences from uncultured organisms. Potentially three novel catechol 1,2-dioxygenases were identified from Bacillus sp. TN102, Gordonia sp. TN103 and Rhodococcus sp. TN112. Highly homologous tautomerase and toluene dioxygenases amongst environmental samples isolated from the contaminated environment suggested horizontal gene transfer while limited success in PCR detection of the other three genes indicates that these isolates may still be a source of novel genes. [Projekat Ministarstva nauke Republike Srbije, br. 173048]
机译:属于微生物芳香族生物降解途径的新基因的鉴定和表征非常重要,因为它们已被证明是多功能生物催化剂。在这项研究中,已使用PCR方法筛选了从下部和上部芳香族生物降解途径中筛选出的五个基因,筛选出能够降解多种芳香族化合物的19种环境细菌菌株。在4-草酰巴豆酸酯互变异构酶和甲苯双加氧酶的情况下,尽管存在于大多数环境分离物中,但是却遇到了非常有限的基因多样性。这些基因在环境样品中的高度保守的序列显示与恶臭假单胞菌属的特征化的相应基因的基因序列高度同源。使用基于已知的邻苯二酚和萘双加氧酶序列的简并引物进行筛选,得到的扩增片段数量有限。仅扩增了来自两个芽孢杆菌分离株的两个儿茶酚2,3-二加氧酶,并且与来自特征生物的双加氧酶没有显着相似性,但是与未培养生物中的部分儿茶酚2,3-二加氧酶序列具有80-90%的同一性。从芽孢杆菌中鉴定出潜在的三种新型儿茶酚1,2-二加氧酶。 TN102,Gordonia sp。 TN103和红球菌TN112。从受污染的环境中分离出的环境样品中高度同源的互变异构酶和甲苯双加氧酶表明水平基因转移,而在PCR检测中其他三个基因的成功有限表明这些分离物可能仍然是新基因的来源。 [Projekat Ministarstva nauke Republike Srbije,br。 173048]

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