Development of Copper-Tolerant Mutant of Soluble Methane Monoxygenase-ProducingMethanotrophs for Application in Bioremediation

摘要

The objective of this research was to create copper-tolerant soluble methanemonooxygenase (sMMO)-producing methanotrophs capable of producing high sMMO activity for the degradation of trichloroethylene (TCE). Two methanotrophic strains were utilized for mutagenesis, Methylosinus trichosporium OB3b and Methylosinus sporium 2C10P. The work utilized three different mutagenesis systems that included dichioromethane mutagenesis; transposon mutagenesis; and UV mutagenesis. The data gathered were the measurement of chlorinated aliphatic transformation, growth kinetics, and calculation of sMMO activity. The dichloromethane mutagenesis method successfully produced copper-tolerant mutagenes. The mutants developed were able to stably express sMMO in the presence of copper levels up to 10 uM at approximately 50-80% of their maximal sMMO levels when cultivated in the absence of copper.