Quantitative Determination of 2-Mercaptoethane Sulphonate as a Biomarker for Methanogens in Soil Using HPLC
All methanogens,irrespective of their preferred carbon sources,form coenzyme M (Co-M:2-mercaptoethane sulphonate) as an intermediate to produce methane (CH4).Generally,a real-time PCR technique is used for indirect quantification of methanogen abundance in soil.But the assay of mcrA genes,coding methyl Co-M reductase enzyme,by real-time PCR is sometimes limited by the necessity of sophisticated laboratory set-up and experienced operators.In this experiment,the method for quantifying Co-M was standardized by HPLC using UV detector and the values of Co-M were compared with CH4 emission flux and methanogen activity of soil.Our objectives were to standardize the Co-M quantification technique and to evaluate its feasibility as a biomarker for methanogens in soil.The Co-M was extracted from soil by rupturing the cell membrane of methanogens using lysis buffer.Trichloroacetic acid (0.5 M) and acetonitrile mixture (70:30,v/v) were used as a mobile phase for measuring Co-M by HPLC at 270 nm.The precision of the method was more than 97% and 90.3% ± 8.1% of the added Co-M standard was recovered from soil by this method.The Co-M concentrations at different rice cultivation stages had shown significant correlations with CH4 emission and methanogen activity in soil.The increases in methanogen activity and Co-M concentration following organic substrate incorporation were proportional to the changes in CH4flux from rice paddy soils.The conversion factor (155.03±14.20μg CH-4 produced mmol-1 Co-M d-1) could be used to calculate methanogen activity from Co-M concentrationsin soil.Based on these results,it is proposed that Co-M can be used as a biomarker for methanogen activity and the above-mentioned method can be used for an easy but precise quantification of Co-M in soil.
Coenzyme M HPLC method standardization Methanogen activity Methanogen biomarker Conversion factor
Prabhat Pramanik Pil Joo Kim
Division of Applied Life Science,Gyeongsang National University,Jinju 660-701,Korea
国际会议
the 16th Meeting of the International Humic Substances Society (第十六届国际腐殖物质学会大会(IHSS 16))
杭州
英文
486-488
2012-09-09（万方平台首次上网日期，不代表论文的发表时间）