Cell-free system for converting methane into fuel and chemical compounds

A cell-free method for large-scale conversion of methane to pyruvate, comprising:

• combining, in a bioreactor, methane, oxygen, and a cell lysate mixture of at least two cell lysates, at least one of which is obtained from recombinant cells, wherein the cell lysate mixture contains the following enzymes:
• a methane monooxygenase, a methanol dehydrogenase, a hexulose-6-phosphate synthase, a 6-phospho-3-hexuloisomerase, a 6-phosphofructokinase, a fructose bisphosphate aldolase, a triose phosphate isomerase, a transketolase, a ribose-5-phosphate isomerase or a ribulose-5-phosphate 3-epimerase, a glyceraldehyde 3-phosphate dehydrogenase, a phosphoglycerate kinase, a phosphoglycerate mutase, an enolase, and a pyruvate kinase, wherein the recombinant cells overexpress at least one of the foregoing enzymes, to form a cell-free reaction mixture;
• and
• incubating under suitable conditions the cell-free reaction mixture to convert methane to pyruvate, thereby producing pyruvate at a production rate of at least 1 g/L-h.