Dong Ming, Gonzalez Tara D, Klems Meghan M, Steinberg Lisa M, Chen Wilfred, Papoutsakis Eleftherios T, Bahnson Brian J
Dept. of Chemistry & Biochemistry, University of Delaware, Newark, DE, 19716.
Dept. of Chemical & Biomolecular Engineering, Delaware Biotechnology Inst., University of Delaware, Newark, DE, 19711.
Biotechnol Prog. 2017 Sep;33(5):1243-1249. doi: 10.1002/btpr.2503. Epub 2017 Jun 12.
Methanol:coenzyme M methyltransferase is an enzyme complex composed of three subunits, MtaA, MtaB, and MtaC, found in methanogenic archaea and is needed for their growth on methanol ultimately producing methane. MtaABC catalyzes the energetically favorable methyl transfer from methanol to coenzyme M to form methyl coenzyme M. Here we demonstrate that this important reaction for possible production of methanol from the anaerobic oxidation of methane can be reversed in vitro. To this effect, we have expressed and purified the Methanosarcina barkeri MtaABC enzyme, and developed an in vitro functional assay that demonstrates MtaABC can catalyze the energetically unfavorable (ΔG° = 27 kJ/mol) reverse reaction starting from methyl coenzyme M and generating methanol as a product. Demonstration of an in vitro ability of MtaABC to produce methanol may ultimately enable the anaerobic oxidation of methane to produce methanol and from methanol alternative fuel or fuel-precursor molecules. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:1243-1249, 2017.
辅酶M甲基转移酶是一种由MtaA、MtaB和MtaC三个亚基组成的酶复合物,存在于产甲烷古菌中,是古菌利用甲醇生长并最终产生甲烷所必需的。MtaABC催化从甲醇到辅酶M的能量有利的甲基转移反应,形成甲基辅酶M。在此我们证明,甲烷厌氧氧化可能产生甲醇的这一重要反应在体外可以逆转。为此,我们表达并纯化了巴氏甲烷八叠球菌的MtaABC酶,并开发了一种体外功能测定法,该方法证明MtaABC可以催化从甲基辅酶M开始的能量不利(ΔG° = 27 kJ/mol)的逆反应,生成甲醇作为产物。证明MtaABC在体外产生甲醇的能力最终可能使甲烷厌氧氧化产生甲醇,并从甲醇生产替代燃料或燃料前体分子。© 2017美国化学工程师学会 生物技术进展,33:1243 - 1249,2017年。