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  • An engineered methanogenic pathway derived from the domains Bacteria and Archaea.

An engineered methanogenic pathway derived from the domains Bacteria and Archaea.

mBio (2010-11-10)
Daniel J Lessner, Lexan Lhu, Christopher S Wahal, James G Ferry
ABSTRACT

A plasmid-based expression system wherein mekB was fused to a constitutive Methanosarcina acetivorans promoter was used to express MekB, a broad-specificity esterase from Pseudomonas veronii, in M. acetivorans. The engineered strain had 80-fold greater esterase activity than wild-type M. acetivorans. Methyl acetate and methyl propionate esters served as the sole carbon and energy sources, resulting in robust growth and methane formation, with consumption of >97% of the substrates. Methanol was undetectable at the end of growth with methyl acetate, whereas acetate accumulated, a result consistent with methanol as the more favorable substrate. Acetate was consumed, and growth continued after a period of adaptation. Similar results were obtained with methyl propionate, except propionate was not metabolized.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Methyl acetate, natural, 98%, FG
Sigma-Aldrich
Methyl acetate, ≥98%, FG
Sigma-Aldrich
Methyl acetate, anhydrous, 99.5%
Supelco
Methyl acetate, analytical standard
Sigma-Aldrich
Methyl acetate, ReagentPlus®, 99%