In recent years, thennostable enzymes from microorganisms have been interested and exploited in their ability applications. In comparison with mesophilic enzymes, thennostable enzymes have dominance, particularly in the manufacturing process which requires high temperature. Protease - the most widely used proteolytic enzyme in the commercial market which includes thennostable proteases have been used in the tenderizing meat process, the treatment process of emission of foul smell on the decomposition of animal carcasses in slaughter-houses, and seafood processing. On the other hand, thanks to the development of NextGeneration sequencing machine, bioinfonnatics techniques, and metagenomic technique which allow researchers could access to genetic resources of unculturable microorganisms and then find out a lot of new species, enzymes, and compounds with biological activity... In this paper, the authors present the creation process of DNA metagenome library from My Lam hot spring with about 30.000 clones were screened on the substrate specificity, and a complete gene encoding the protease was obtained from this library. Protease gene sequencing showed that the gene has a length of 2453 bp which contained an open reading frame of 2106 nucleotides that encoded for 701 amino acids with 92 percent homology compared with sequences of thennostable islandicum trom Fervidobacterium islandisin. Thus, it can be seen clearly that the advantages of using metagenome techniques might allows us to receive target genes which were obtained trom hardly isolated microorganisms by common methods.