Oligotrophic Gene Expression in Rhodococcus erythropolis N9T-4 under Various Nutrient Conditions

Abstract

Rhodococcus erythropolis N9T-4 is a super oligotroph that grows on an inorganic basal medium without any additional carbon and nitrogen sources and requires CO2 for its oligotrophic growth. Previously, we found that two genes, aldA and mnoA, encoding NAD-dependent aliphatic aldehyde dehydrogenase and N,N′-dimethyl-4-nitrosoaniline-dependent methanol dehydrogenase, respectively, were highly upregulated under oligotrophic conditions. In this study, we constructed reporter plasmids containing an enhanced green fluorescent protein gene under aldA or mnoA promoters (pAldA and pMnoA, respectively). Fluorescence analysis of N9T-4 cells with reporter plasmids revealed that tryptone and yeast extract strongly repressed the expression of oligotrophy-connected genes, whereas the effect of casamino acids was moderate. Furthermore, remarkably high expression of aldA and mnoA was observed when the reporter strains were grown in media containing primary alcohols, particularly ethanol. Malic acid repressed ethanol-induced gene expression, suggesting that C2 metabolism is involved in the oligotrophic growth of N9T-4. The regulation of oligotrophic gene expression elucidated in this study could provide appropriate conditions for the production of useful compounds in an oligotrophic microbial process.