Department of Environmental Engineering, INHA University, Incheon, Republic of Korea.
Appl Biochem Biotechnol. 2013 Nov;171(6):1487-99. doi: 10.1007/s12010-013-0410-0. Epub 2013 Aug 21.
Using a methanotrophic consortium (that includes Methylosinus sporium NCIMB 11126, Methylosinus trichosporium OB3b, and Methylococcus capsulatus Bath) isolated from a landfill site, the potential for partial oxidation of methane into methanol through selective inhibition of methanol dehydrogenase (MDH) over soluble methane monooxygenase (sMMO) with some selected MDH inhibitors at varied concentration range, was evaluated in batch serum bottle and bioreactor experiments. Our result suggests that MDH activity could effectively be inhibited either at 40 mM of phosphate, 100 mM of NaCl, 40 mM of NH4Cl or 50 μM of EDTA with conversion ratios (moles of CH3OH produced per mole CH4 consumed) of 58, 80, 80, and 43 %, respectively. The difference between extent of inhibition in MDH activity and sMMO activity was significantly correlated (n = 6, p < 0.05) with resultant methane to methanol conversion ratio. In bioreactor study with 100 mM of NaCl, a maximum specific methanol production rate of 9 μmol/mg h was detected. A further insight with qPCR analysis of MDH and sMMO coding genes revealed that the gene copy number continued to increase along with biomass during reactor operation irrespective of presence or absence of inhibitor, and differential inhibition among two enzymes was rather the key for methanol production.
利用从垃圾填埋场中分离出的甲烷营养菌混合体(包含嗜甲基菌(Methylosinus sporium)NCIMB 11126、三氯甲烷甲烷杆菌(Methylosinus trichosporium)OB3b 和荚膜甲基球菌(Methylococcus capsulatus)Bath),通过选择性抑制甲醇脱氢酶(MDH)对可溶性甲烷单加氧酶(sMMO)的抑制,在分批血清瓶和生物反应器实验中评估了甲烷通过部分氧化生成甲醇的潜力。我们的结果表明,在 40 mM 磷酸盐、100 mM 氯化钠、40 mM 氯化铵或 50 μM EDTA 等不同浓度的几种 MDH 抑制剂的作用下,MDH 活性可以有效地被抑制,转化率(生成的甲醇摩尔数与消耗的甲烷摩尔数之比)分别为 58%、80%、80%和 43%。MDH 活性和 sMMO 活性抑制程度的差异与甲烷到甲醇的转化率呈显著相关(n = 6,p < 0.05)。在 100 mM 氯化钠的生物反应器研究中,检测到最大的甲醇比生成速率为 9 μmol/mg h。进一步通过 qPCR 分析 MDH 和 sMMO 编码基因的结果表明,在反应器运行过程中,无论是否存在抑制剂,基因拷贝数都随着生物质的增加而持续增加,两种酶之间的差异抑制是甲醇生产的关键。