Diversity of hydrolase-producing halophilic bacteria and evaluation of their enzymatic activities in submerged cultures

Abstract

Purpose: In this work, we assessed the diversity of culturable halophilic bacteria that produce hydrolytic enzymes from both natural and artificial hypersaline regions in the pre-Rif region of Morocco. Methods: Bacteria were isolated from three hypersaline sites, in solid medium supplemented with various salt concentrations ranging from 0 to 330 g/L. Physical and chemical characteristics of samples from the isolation site were determined to suggest eventual correlations with the occurrence of the halophilic bacteria. Assays on enzymatic activities were performed in submerged cultures in the presence of various salt concentrations and appropriate substrates. Results: Out of a collection of 227 halophilic bacteria, four halophilic groups were established as slightly halophilic, moderately halophilic, halotolerant, or extremely halophilic, with a predominance of halophilic bacteria in the natural hypersaline sites compared to the artificial one. Within this collection, 189 strains showed important hydrolytic activities in submerged cultures with enzymatic activities up to 76 U/mg. Strain characterization and identification was based on phenotypic and molecular traits and allowed the identification of at least 26 genera including Bacillus, Chthonibacter, Mariniabilia, Halobacillus, Salinococcus, Cerasicoccus, Ulvibacter, Halorubrum, Jeatgalicoccus, Brevibacterium, Sanguibacter, Shewanella, Exiguobacterium, Gemella, and Planomicrobium. Conclusion: Data from this study give insights about the origin and the occurrence of halophilic bacteria in natural hypersaline environments compared to artificial hypersaline sites. The occurrence of halophilic hydrolase enzymes from halophilic bacteria gives insights to different applications in biotechnology, thanks to their ability to produce adaptive enzymes and survival strategies to overcome harsh conditions.