AlmA involved in the long-chain -alkane degradation pathway in ADP1 is a Baeyer-Villiger monooxygenase.

Many species can grow on -alkanes of varying lengths (≤C40). AlmA, a unique flavoprotein in these strains, is the only enzyme proven to be required for the degradation of long-chain (LC) -alkanes, including C32 and C36 alkanes. Although it is commonly presumed to be a terminal hydroxylase, its role in -alkane degradation remains elusive. In this study, we conducted physiological, biochemical, and bioinformatics analyses of AlmA to determine its role in -alkane degradation by ADP1. Consistent with previous reports, gene deletion analysis showed that was vital for the degradation of LC -alkanes (C26-C36). Additionally, enzymatic analysis revealed that AlmA catalyzed the conversion of aliphatic 2-ketones (C10-C16) to their corresponding esters, but it did not conduct -alkane hydroxylation under the same conditions, thus suggesting that AlmA in strain ADP1 possesses Baeyer-Villiger monooxygenase (BVMO) activity. These results were further confirmed by bioinformatics analysis, which revealed that AlmA was closer to functionally identified BVMOs than to hydroxylases. Altogether, the results of our study suggest that LC -alkane degradation by strain ADP1 possibly follows a novel subterminal oxidation pathway that is distinct from the terminal oxidation pathway followed for short-chain -alkane degradation. Furthermore, our findings suggest that AlmA catalyzes the third reaction in the LC -alkane degradation pathway.IMPORTANCEMany microbial studies on -alkane degradation are focused on the genes involved in short-chain -alkane (≤C16) degradation; however, reports on the genes involved in long-chain (LC) -alkane (>C20) degradation are limited. Thus far, only AlmA has been reported to be involved in LC -alkane degradation by spp.; however, its role in the -alkane degradation pathway remains elusive. In this study, we conducted a detailed characterization of AlmA in ADP1 and found that AlmA exhibits Baeyer-Villiger monooxygenase activity, thus indicating the presence of a novel LC -alkane biodegradation mechanism in strain ADP1.