• Journal of environmental and Sanitary engineering
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• v.23 no.2
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• pp.1-18
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• 2008

The tetrachloroethylene (PCE) dehalogenase of Clostridium bifermentans DPH-1 (a halorespiring organism) was purified, cloned, and sequenced. This enzyme is a homodimer with a molecular mass of ca. 70 kDa and exhibits dehalogenation of dichloroethylene isomers along with PCE and trichloroethylene (TCE). Broad range of substrate specificity for chlorinated aliphatic compounds (PCE, TCE, cis-1,2-dichloroethylene, trans-1,2-dichloroethylene, 1,1-dichloroethylene, 1,2-dichloropropene, and 1,1,2-trichloroethane) for this enzyme was also observed. A mixture of propyl iodide and titanium citrate caused a light-reversible inhibition of enzymatic activity suggesting the involvement of a corrinoid cofactor. A partial sequence (81 bp) of the encoding gene for PCE dehalogenase was amplified and sequenced with degenerateprimers designed from the N-terminal sequence (27 amino acid residues). Southern analysis of C. bifermentans genomic DNA using the polymerase chain reaction product as a probe revealed restriction fragment bands. A 5.0 kb ClaI fragment, harboring the relevant gene (designated pceC) was cloned (pDEHAL5) and the complete nucleotide sequence of pceC was determined. The gene showed homology mainly with microbial membrane proteins and no homology with any known dehalogenase, suggesting a distinct PCE dehalogenase. So, C. bifermentans could play some important role in the initial breakdown of PCE and other chlorinated aliphatic compounds in sites contaminated with mixtures of halogenated substances.

• Journal of Environmental Health Sciences
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• v.26 no.2
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• pp.14-21
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• 2000

DEAE-Toyopearl 650S, Superdex pg-75, Poros HQ, Superdex H200의 각종 칼러크로마토그래피를 이용하여 C.bifermentans DPH-1균주로부터 테트라클로로에틸렌(PCE) 분해 효소를 정제했다. 이 PCE 분해효소 (PCE dehalogenase)는 PCE를 환원적 탈염소화 반응에 의해 시스디클로로에딜렌 (cis-1,2-dichloroethylene)에 전환 가능하여, 이 때의 Vmax 및 Km 치는 각각 73 nmol/h.mg protein, 12$\mu$M이었다. 본 PCE dehalogenase의 겔여과 분자량 Maker Kit를 이용한 분석결과(70kDa)는 SDS-PAGE에 나타난 분자량(35kDa)의 약 2배인 것으로 확인되었다. 따라서 본 효소는 분자량 70kDa의 이량체(Homo dimer)인 것으로 추정되었다. 본 효소의 최적온도 및 pH는 각각 35$^{\circ}C$ 및 8.0 이었다. 또한 본 효소는 PCE외의 트리클로로에틸렌, 디클로로에틸렌 이성체, 1,2-디클로로에템, 1,2-디클로로프로판, 1,1,2-트리클로로에탄에 대하여도 활성을 타나내었다. N-말단 아미노산 분석결과에서도 본 효소는 현재 알려진 PCE dehalogenase와 그 배열이 전혀 다른 것으로 나타나 각종 유기염소 화합물의 분해능을 보유한 신종의 PCE 분해효소인 것이 확인되었다.

• Korean Journal of Microbiology
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• v.52 no.2
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• pp.157-165
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• 2016

Under flooded rice fields, methanogens produce methane which comes out through rice stalks, thus rice fields are known as one of the anthropogenic sources of atmospheric methane. Studies have shown that use of manure increases amount of methane emission from rice. To investigate mechanisms by which manure boosts methane emission, comparative soil metagenomics between inorganically (NPK) and pig manure fertilized paddy soils (PIG) were conducted. Results from taxonomy analysis showed that more abundant methanogens, methanotrophs, methylotrophs, and acetogens were found in PIG than in NPK. In addition, BLAST results indicated more abundant carbohydrate mabolisetm functional genes in PIG. Among the methane metabolism related genes, PIG sample showed higher abundance of methyl-coenzyme M reductase (mcrB/mcrD/mcrG) and trimethylamine-corrinoid protein Co-methyltransferase (mttB) genes. In contrast, genes that down regulate methane emission, such as trimethylamine monooxygenase (tmm) and phosphoserine/homoserine phosphotransferase (thrH), were observed more in NPK sample. In addition, more methanotrophic genes (pmoB/amoB/mxaJ), were found more abundant in PIG sample. Identifying key genes related to methane emission and methane oxidation may provide fundamental information regarding to mechanisms by which use of manure boosts methane emission from rice. The study presented here characterized molecular variation in rice paddy, introduced by the use of pig manure.