Veeranagouda Y, Emmanuel Paul P V, Gorla P, Siddavattam D, Karegoudar T B
Department of Biochemistry, Gulbarga University, Karnataka, India.
Appl Microbiol Biotechnol. 2006 Jul;71(3):369-75. doi: 10.1007/s00253-005-0157-9. Epub 2005 Oct 7.
A bacterial strain DGVK1 capable of using N,N-dimethylformamide (DMF) as sole source of carbon and nitrogen was isolated from the soil samples collected from the coalmine leftovers. The molecular phylogram generated using the complete sequence of 16S rDNA of the strain DGVK1 showed close links to the bacteria grouped under Brucellaceae family that belongs to alphaproteobacteria class. Specifically, the 16S rDNA sequence of strain DGVK1 has shown 97% similarity to Ochrobactrum anthropi LMG 3331 (D12794). This bacterium has also shown impressive growth on dimethylamine, methylamine, formaldehyde and formate that are considered to be the prominent catabolic intermediates of DMF. DMF degradation has led to the accumulation of ammonia and dimethylamine contributing to the increase of pH of the medium. The DMF-grown resting cells of Ochrobactrum sp. DGVK1 have also contributed for the release of ammonia when resting cell suspension was added to phosphate buffer containing DMF. Similar experiments done with the glucose-grown cultures have not produced ammonia and thus indicating the inducible nature of DMF-degrading enzymes in Ochrobactrum sp. DGVK1. Further, dimethylformamidase, dimethylamine dehydrogenase and methylamine dehydrogenase, the key enzymes involved in the degradation of DMF, were assayed, and the activities of these enzymes were found only in DMF-grown cultures further confirming the inducible nature of the DMF degradation. Based on these results, DMF degradation pathway found in Ochrobactrum sp. DGVK1 has been proposed.
从煤矿废弃物采集的土壤样本中分离出了一种能够将N,N-二甲基甲酰胺(DMF)作为唯一碳源和氮源的细菌菌株DGVK1。使用菌株DGVK1的16S rDNA完整序列生成的分子系统发育树显示,它与属于α-变形菌纲布鲁氏菌科的细菌有密切联系。具体而言,菌株DGVK1的16S rDNA序列与嗜人苍白杆菌LMG 3331(D12794)显示出97%的相似性。该细菌在二甲胺、甲胺、甲醛和甲酸盐上也表现出令人印象深刻的生长,这些被认为是DMF的主要分解代谢中间体。DMF降解导致氨和二甲胺的积累,促使培养基pH值升高。当将嗜油杆菌属DGVK1的以DMF培养的静息细胞悬液加入含有DMF的磷酸盐缓冲液中时,也会释放出氨。对以葡萄糖培养的培养物进行的类似实验未产生氨,因此表明嗜油杆菌属DGVK1中DMF降解酶具有诱导性。此外,还测定了参与DMF降解的关键酶——二甲基甲酰胺酶、二甲胺脱氢酶和甲胺脱氢酶,发现这些酶的活性仅存在于以DMF培养的培养物中,进一步证实了DMF降解的诱导性。基于这些结果,提出了嗜油杆菌属DGVK1中的DMF降解途径。