1An extremely thermophilic bacterium, Geobacillus thermoleovorans B23, is capable of 2 degrading a broad range of alkanes (with carbon chain lengths ranging between C11 and 3 C32) at 70 °C. Whole--genome sequence analysis revealed that unlike most alkane--degrading 4 bacteria, strain B23 does not possess an alkB--type alkane monooxygenase gene. Instead, it 5 possesses a cluster of three ladA--type genes, ladAα B23 , ladAβ B23 , and ladB B23 , on its 6 chromosome, whose protein products share significant amino acid sequence identities, 49.8, 734.4, and 22.7%, respectively with that of ladA alkane monooxygenase gene found on a 8 plasmid of Geobacillus thermodetrificans NG 80--2. Each of the three genes, ladAα B23 , 9 ladAβ B23 , and ladB B23 , was heterologously expressed individually in an alkB1 deletion mutant 10 strain, Pseudomonas fluorescens KOB2Δ1. It was found that all three genes were functional 11 in P. fluorescens KOB2Δ1, and partially restored alkane degradation activity. In this study, 12we suggest that G. thermoleovorans B23 utilizes multiple LadA--type alkane 13 monooxygenases for the degradation of a broad range of alkanes. 14 15