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Microbiology and Biotechnology Letters

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Environmental Microbiology (EM)  |  Microbial Ecology and Diversity

Microbiol. Biotechnol. Lett. 2024; 52(1): 76-87

https://doi.org/10.48022/mbl.2401.01010

Received: January 19, 2024; Revised: February 29, 2024; Accepted: March 7, 2024

Bioprospecting of Culturable Halophilic Bacteria Isolated from Mediterranean Solar Saltern for Extracellular Halotolerant Enzymes

Ahmed Mohamed Ali1, Tahany M.A. Abdel-Rahman1, and Mohamed G. Farahat1,2*

1Botany and Microbiology Department, Faculty of Science, Cairo University, Giza 12613, Egypt
2Biotechnology Department, Faculty of Nanotechnology for Postgraduate Studies, Sheikh Zayed Branch Campus, Cairo University, Sheikh Zayed City, Giza 12588, Egypt

Correspondence to :
Mohamed  G. Farahat,          farahat@cu.edu.eg

Abstract

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Halophilic bacteria are promising reservoirs for halotolerant enzymes that have gained much attention in biotechnological applications due to their remarkable activity and stability. In this study, 62 halophilic bacterial strains isolated from a solar saltern were screened for the production of various extracellular enzymes. The results revealed that 31 strains (50%) were positive for amylase production while 26 strains (41.9%) were positive for protease. Further, 22 strains (35.48%) exhibited β-glucosidase activity and only 17 (27.41%) demonstrated lipase activity. Of the investigated halophiles, ten strains growing in the presence of ≥15% NaCl (w/v) were selected and identified based on their 16S rRNA gene sequences as Halomonas meridiana, Salinivibrio costicola, Virgibacillus oceani, Virgibacillus marismortui, Marinobacter lipolyticus, Halobacillus karajensis, Salicola salis, Pseudoalteromonas shioyasakiensis, Salinicoccus amylolyticus, and Paracoccus salipaludis. Therefore, the present study highlights the diversity of the culturable halophilic bacteria in a Mediterranean solar saltern, harboring various valuable halotolerant enzymes.

Keywords: Halophilic bacteria, solar salterns, hydrolytic enzymes, bacterial diversity

Graphical Abstract


Introduction

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Microorganisms have various adaptive strategies to tolerate environmental stresses, especially extremophiles which can survive under adverse conditions, such as high temperature, pH, and salinity [1]. This can be achieved by many mechanisms including the ability to produce particular secondary metabolites, which have many applications in various industrial fields and biotechnological research [2]. Extremophilic microorganisms that are capable of surviving in high salinity can grow in habitats such as the Dead Sea, solar salterns, and salt lakes. These organisms can produce extracellular enzymes with potential interest in several biotechnological and industrial applications due to their high activity and stability at low water levels [1, 3, 4]. It was reported that halophiles-derived enzymes were active under extreme conditions because their surfaces carry a high number of amino acid residues of negative charges [5]. Hydrolytic enzymes such as proteases, lipases, β-glucosidase, inulinases, cellulases, and amylases received increased attention due to their significant industrial roles in wastewater treatment, biosynthetic processes, agriculture, medicine, foods, detergent, textile, paper, and biofuel industries [6−10]. Also, hydrolytic enzymes have crucial roles in bioremediation processes and biodegradation of pollutants [11−14]. In this context, various bacteria strains isolated from marine habitats were reported as promising sources for a wide range of significant commercial hydrolytic enzymes including cellulases, nucleases, inulinases, xylanases, amylases, galactosidases, glucosidases, lipases, chitinases, dextranases, proteases, etc. [15−20]. Amylases are classified as hydrolyzing enzymes that hydrolyze starch into monosaccharide units by acting on its α-1,4-glucosidic linkages [21, 22]. Various amylases were applied in industrial sectors for starch liquefication, biomasses saccharification, fabrics desizing, beverage fermentation, and syrups production [23, 24]. In this regard, α-amylase from the marine bacterium Bacillus subtilis S8-18 was used to inhibit/disrupt the biofilms of some human bacterial pathogens such as against methicillin-resistant Staphylococcus aureus (MRSA) and Vibrio cholerae [25]. Also, α-amylase from halophilic Streptomyces sp. has been used for hydrolysis of agro-wastes [26]. The enzyme derived from Bacillus tequilensis TB5 is used as a laundry additive and as a textile desizer [27]. Microorganisms that are isolated from environments with high salt concentrations have been reported as a prolific source for the production of amylases such as Halomonas meridiana [28] and Zunongwangia profunda [29]. In addition, proteases are one of the economically important enzymes involved in the hydrolysis of proteins into small peptides and/or free amino acids by acting on peptide linkages [30]. These enzymes have many applications in the leather industry, textiles, food processing, and dairy production [31, 32].

Furthermore, the halophilic Bacillus sp. strain SP II-4 has been reported as a significant producer of alkaline protease which demonstrated better content lens cleansing efficiency along with common chemical disinfectant [33]. Protease from a marine bacterium, Bacillus tequilensis P15, has industrial applications such as removing of blood stains from fabrics, stripping off the gelatin from used photographic films, and dehairing of hide [34]. Also, the halotolerant protease produced by Halobacillus andaensis has been used for the production of bioactive peptides from fish muscle protein [35]. Halotolerant proteases have been reported to be able to remove recalcitrant blood and egg stains, and are used in the leather industry as dehairing agents at high salt conditions [36]. Microorganisms from hypersaline habitats have been reported as a significant source of protease, such as Halococcus agarilyticus [37] and Lysinibacillus fusiformis sp. [38]. Furthermore, lipases that hydrolyze fat into glycerol and fatty acids play a significant role in various industrial fields such as biodiesel, foods and drinks, leather, detergents, and pharmaceuticals [39]. Marinederived lipases exhibited high activity under adverse conditions, so marine microorganisms have been reported as prolific sources for producing marine lipases [40−42]. Lipase derived from halophilic microorganisms has been reported as an prominent enzyme in food industries, including baking and brewing, omega-3 production, natural sweeteners, and food processing [43]. Lipase from Staphylococcus warneri is used in fish sauce production to enhance the flavor [44]. In addition, lipase derived from Marinobacter lipolyticus SM19 has been used in the pharmaceuticals and food industry through the synthesis of polyunsaturated fatty acids from fish oil [45]. In the medical field, a halothermostable lipase from Oceanobacillus sp. PUMB02 was used to disrupt biofilm against different food pathogens such as Listeria sp., Bacillus sp., Serratia sp., and Vibrio parahemolyticus [46]. It has been suggested that halotolerant lipases are promising candidates to be applied in bioremediation processes. Cold active lipases from Pseudoalteromonas sp. are used in the bioremediation of aqueous systems contaminated with oil and in detergent formulations [47], and lipase from Halomonas sp. C2SS100 showed the ability to remove of lubricating oil stains from polycotton [48]. Many halophilic bacteria have been documented as promising sources for the production of lipases such as Halobacillus truperi [49] and Bacillus amyloliquefaciens [50]. Moreover, β-glucosidase is a hydrolyzing enzyme that catalyzes the release of nonreducing terminal glucosidic residues from glycosylated metabolites or oligosaccharides and has various applications in the food, chemical, and pharmaceutical industries for the production of Gardinia Blue (natural pigment), gentiooligosaccharides, and bioconversion of polydatin to resveratrol [51]. In the food industry, β- glucosidases maintain the food flavor [52−54]. Halophilic β-glucosidase from Pseudoalteromonas has been used in industrial production and food processing due to its significant activity toward isoflavones [55]. Also, this enzyme is involved in the synthesis of various compounds like glycoconjugates and oligosaccharides [56]. In the present study, we addressed the diversity of culturable halophilic bacteria isolated from Mediterranean solar saltern and investigated their ability to produce various extracellular hydrolytic enzymes.

Material and Methods

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Sample Collection

Sediments and brine samples were collected from a solar saltern, in Egypt (30°59'24.9"N 29°37'00.4"E) in sterile plastic bags and transferred to the laboratory through a short time after sampling and surveyed immediately for bacterial isolation.

Isolation of Halophilic Bacteria

Isolation of halophilic bacteria was performed using tryptic soy agar medium (TSA, Condalab, Spain) supplemented with 10% (w/v) NaCl. In brief, 100 μl of each collected sample were plated and the inoculated plates were incubated at 37℃ and checked daily for up to 7 days. The recovered colonies were transferred on new agar plates and sub-cultured many times until pure cultures were obtained. For the preservation of bacterial isolates, stocks of 20% glycerol of pure cultures were prepared and stored at -80℃.

Screening for Extracellular Hydrolytic Enzymes

Amylase activity. The potential of halophilic isolates for amylase production was investigated using starch agar medium containing (g/l): yeast extract, 1; peptone, 5; soluble starch, 2; agar, 20; supplemented with 10% NaCl. After incubation at 37℃ for 3−5 days, plates were flooded with iodine-potassium iodide solution. The formation of a clear zone around the bacterial colonies was recorded as starch hydrolysis [57].

Protease activity. The proteolytic activity was assessed by the method described by [57] with some modifications. Each strain was inoculated on skim milk agar medium containing (g/l): skim milk, 20; glucose, 1; tryptone, 5; yeast extract, 2.5; agar, 20; supplemented with 10% NaCl. After incubation at 37℃ for 5 days, the plates were checked. The appearance of a clear zone around indicates protease activity.

Lipase activity. The investigation of lipase activity was conducted using a screening medium containing (g/l): peptone, 10; CaCl2·2H2O, 0.1; agar, 20, supplemented with 10% NaCl (w/v). After autoclaving, sterile tween 80 was added to a final concentration of 1% (v/v). Each bacterial strain was inoculated and the plates were incubated at 37℃ for 4−5 days. Subsequently, the appearance of white precipitate around the bacterial growth indicated the lipase activity [58].

β-Glucosidase activity. The β-glucosidase activity was inspected using TSA medium containing (g/l): beef extract, 3; peptone, 5; agar, 20, supplemented with 10% NaCl, 0.05% esculin and 0.01% ferric citrate. The formation of a brown-black zone around the bacterial colonies after incubation at 37℃ for 5 days indicated β-glucosidase activity [51].

Salinity Tolerance

All isolated strains were inoculated in tryptic soy broth supplemented with different concentrations of NaCl (10, 15, 20 and 25%) and incubated at 37℃ in a shaking incubator (140 rpm). The cultures were checked daily for bacterial growth up to 15 days.

Molecular Identification and Phylogenetic Analysis

Extraction of genomic DNA from bacterial isolates was performed by using GeneJET™ Genomic DNA Purification Kit according to the manufacturer protocol. Amplification of nearly full-length 16S rRNA gene was conducted by PCR using universal primers, 27F (AGAGTTTGATCMTGGCTCAG) and 1492R (TACGGYTACCTTGTTACGACTT) [59]. Afterward, the amplicons were purified and sequenced using ABI 3730xI sequence analyzer (Applied Biosystems, USA), and the sequences were analyzed using the Basic Local Alignment Search Tool (http://www.ncbi.nlm.nih.gov/blast). Construction of the phylogenetic tree was performed using the Neighborjoining method based on bootstrap values (1000 replications with MEGAX software). The sequences of the 16S rRNA gene were deposited in the GenBank.

Results

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Isolation of Halophilic Bacteria

In the present study, 62 halophilic bacterial strains were isolated on TSA medium supplemented with 10% NaCl, from a solar saltern, in Egypt. After that, all isolated halophiles were screened for the production of various extracellular hydrolytic enzymes.

Screening of Extracellular Hydrolytic Enzymes

All isolated bacterial halophiles were screened for their ability to produce four extracellular hydrolytic enzymes (amylase, protease, lipase, and β-glucosidase) in the presence of 10% NaCl. Of the investigated strains, 54/62 (87.09%) were positive for at least one extracellular enzyme. On the other hand, eight strains (12.91%) did not produce any of the investigated extracellular enzymes. Results showed that 31 strains (50%) had amylase activity (Fig. 1A). Regarding the protease activity, 26 strains (41.9%) exhibited proteolytic activity on skimmed milk agar plates (Fig. 1B). Seventeen strains (27.41%) showed lipase activity (Fig. 1C) and 22 halophilic strains (35.48%) were found to produce dark brown zones around their colonies, indicating β-glucosidase activity (Fig. 1D). Results revealed that only three bacterial strains were positive for all investigated enzymes and seven strains produced three different enzymes. Moreover, 19 bacterial strains exhibited positive activity for two investigated enzymes, and 25 strains exhibited activity for only one enzyme (Fig. 2).

Figure 1.Screening of extracellular enzyme production. (A) amylase, (B) protease, (C) lipase, and (D) β-galactosidase of the selected halophilic bacteria.

Figure 2.Venn diagram shows prevalence of extracellular amylase, protease, lipase, and β-galactosidase.

Salinity Tolerance

Bacterial growth in different concentrations of NaCl (10−25%, w/v) was investigated. All the investigated isolates exhibited significant growth at 10% NaCl. Results revealed that three strains (SHB1, SHB21 and SHB59) were able to grow at NaCl concentrations up to 25%, while SHB7, SHB26 and SHB37 exhibited growth up to 20% NaCl. Three strains (SHB13, SHB22, SHB55 and SHB60) showed growth up to 15% NaCl (Table 1). Among 62 halophilic strains, 10 exhibiting the ability to grow in high salt concentrations (≥15%) were selected while 52 strains that did not grow on more than 10% NaCl were maintained at -80℃ for future investigations. The selected strains were subjected to molecular identification based on 16S rRNA analysis.

Table 1 . Salinity tolerance of halophilic strains isolated from a Mediterranean solar saltern located at the northern coast of Egypt.

StrainGrowth with NaCl (%, w/v)
10152025
SHB1++++
SHB7+++-
SHB13++--
SHB21++++
SHB22++--
SHB26+++-
SHB37+++-
SHB55++--
SHB59++++
SHB60++--

‘+’ positive for the test, ‘-’ negative for the test.

Molecular Identification and Phylogenetic Analysis

Based on 16S rRNA gene analysis, the selected halophilic strains were identified as Halomonas meridiana, Salinivibrio costicola, Virgibacillus oceani, Virgibacillus marismortui, Marinobacter lipolyticus, Halobacillus karajensis, Salicola salis, Pseudoalteromonas shioyasakiensis, Salinicoccus amylolyticus and Paracoccus salipaludis. Sequences of the 16S rRNA gene were submitted to the GenBank and the accession numbers were assigned (Table 2). The Neighbor-joining phylogenetic tree was constructed based on 16S rRNA gene sequences of the bacterial isolates (Fig. 3).

Table 2 . Identification of extreme halophilic bacterial strains based on 16S rRNA gene sequence analysis.

StrainAccession NumberClosest SpeciesSimilarity (%)
SHB1PP035898Halomonas meridiana99.93
SHB7OR616804Salinivibrio costicola99.30
SHB13OR616805Virgibacillus oceani99.79
SHB21OR616806Virgibacillus marismortui99.87
SHB22OR616807Marinobacter lipolyticus99.86
SHB26OR616808Halobacillus karajensis99.79
SHB37OR616809Salicola salis99.64
SHB55OR616810Pseudoalteromonas shioyasakiensis99.78
SHB59OR616811Salinicoccus amylolyticus99.55
SHB60OR616812Paracoccus salipaludis99.77


Figure 3.Neighbor-joining phylogenetic tree based on 16S rRNA gene sequences showing the relationship between the selected strains and the most closely related species.

Concerning the selected bacterial strains, results revealed that ten identified strains activity of possess at least one hydrolytic enzyme (Table 3). The most active strain that exhibited activity of all tested enzymes was H. karajensis SHB26, while M. lipolyticus SHB22 produced lipase only. Furthermore, V. oceani SHB13, S. salis SHB37, and P. shioyasakiensis SHB55 exhibited the activity of 3 enzymes, while H. meridiana SHB1, S. costicola SHB7, V. marismortui SHB21, S. amylolyticus SHB59, and P. salipaludis SHB60 secreted two enzymes.

Table 3 . Extracellular hydrolytic enzymes of extreme halophilic bacteria.

StrainExtracellular Enzyme
AmylaseProteaseLipaseβ-Glucosidase
H. meridiana SHB1+-+-
S. costicola SHB7+-+-
V. oceani SHB13-+++
V. marismortui SHB21-+-+
M. lipolyticus SHB22--+-
H. karajensis SHB26++++
S. salis SHB37+++-
P. shioyasakiensis SHB55++-+
S. amylolyticus SHB59+--+
P. salipaludis SHB60-++-

‘+’ positive for the test, ‘-’ negative for the test.

Discussion

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Recently, halophilic bacteria have been regarded as prolific sources of novel economically important stable enzymes that are functional under extreme conditions. Due to their unique properties, halophilic-derived enzymes are extensively used in many industrial fields. In the present work, 62 halophilic bacteria were isolated from a solar saltern in Egypt among them ten halophilic strains that can grow at NaCl concentrations equal to or more than 15% belonging to nine genera were selected. Because of operating at high salt concentrations with the reducing water activity, enzymes from the extreme halophilic microorganisms exhibit extraordinary stability and are thought to be robust biocatalysts in aqueous/ organic and nonaqueous media with superior activity in various biotechnological applications [60, 61]. Solar salterns are one of the natural habitats that are characterized by the presence of high salt concentrations and have been reported as a rich source for the isolation of halophilic microorganisms. In a similar study, various halophilic bacteria have been isolated from solar salterns in Tunisia, including Halomonas sp., Salinivibrio sp., Cobetia sp., Idiomarina sp., and Marinobacter [62]. Also, 28 halophilic bacteria have been isolated from Indian solar salterns which belonged to seven genera: Stenotrophomonas, Staphylococcus, Bacillus, Enterobacter, Pseudomonas, Oceanobacillus and Ochrabactrum [63] and ten bacterial halophiles have been isolated from hypersaline habitats in Morocco, nine bacterial strains belonged to Halomonas genus and one bacterial strain belonged to Marinobacter genus [64]. Furthermore, 17 bacterial isolates have been isolated from a hypersaline lake in the Northwest of Algeria, which belong to three genera: Virgibacillus, Bacillus, and Halomonas [65]. Moreover, Providencia stuatrti sp. and Lysinibacillus fusiformis sp. were isolated from high-salt environments [38]. From saltern ponds in Trapani, Sicily, Halomonas sp., Salinibacter ruber, and Brevibacterium sp. have been isolated [66]. Various bacterial and archaeal halophiles were isolated from a hypersaline lake in the south of Tunisia, and identified as Salicola sp., Bacillus sp., Halorubrum sp., Natrinema sp., Haloterrigena sp. [67], and Pseudoalteromonas sp. DL-6 has been isolated from marine sediments in the Chinese Bohai Sea [68]. Bacterial isolates related to genera: Brevibacterium, Chromohalobacter, Virgibacillus, Halomonas, Salinivibrio, Nesiotobacter, Cobetia, and Salinicoccus have been isolated from solar salterns in Bulgaria [69]. In a similar study that investigated the halophilic bacteria from a solar saltern located in the Mediterranean Sea (Sfax, Tunisia), 40 halophilic isolates were identified based on phylogenetic analysis of 16S rRNA gene sequences. Their diversity showed that 4 strains (10%) were belonging to Firmicutes and 36 strains (90%) were belonging to Gamma-Proteobacteria. The Gamma-Proteobacteria consisted of various subgroups of the Alcanivoracaceae (5%), the Idiomarinaceae (7.5%), the Alteromonadaceae (10%), the Vibrionaceae (15%) and the Halomonadaceae (52.5%) [70]. In agreement with our results, halophilic bacteria such as Halomonas, Salicola, Halovibrio, and Salinibacter have been isolated from solar salterns in Baja California, Mexico [71]. Also, Marinococcus, Salinibacter, and Cytophaga were isolated from solar salterns of Tamil Nadu, India [72]. Furthermore, similar halophilic bacteria such as Salinibacter ruber, Brevibacterium sp. and Halomonas sp. have been isolated from saltern ponds of Trapani, Sicily [66], and Salinivibrio kushneri has been isolated from salterns, Spain [73]. Although many species belonging to the recovered genera have been described in previous studies, our findings spotlighted the diversity of these strains with emphasis on their extracellular enzymatic activity.

In the present study, amylase was the most prevalent enzyme followed by protease, while lipase was the least prevalent enzyme. The ten extreme halophilic identified strains showed at least one activity of the tested enzymes. The most active strain that showed activity of the four enzymes was H. karajensis, while M. lipolyticus produced lipase only. Meanwhile, V. oceani, S. salis, and P. shioyasakiensis exhibited the activity of three enzymes, but isolates such as H. meridiana, S. costicola, V. marismortui, S. amylolyticus, and P. salipaludis exhibited activity of two enzymes. Similarly, Salicola sp., Bacillus sp., Halorubrum sp., Natrinema sp., and Haloterrigena sp. isolated from hypersaline lake in the south of Tunisia, were screened for the activity of hydrolytic enzymes, including protease, amylase, cellulase, lipase, xylanase, and pectinase and found that most these isolates exhibited significant activity of the tested enzymes [67]. It has been suggested that marine halophilic microorganisms, classified as one class of extremophiles, are an extraordinary source of new enzymes involved in the bioremediation of toxic pollutants and dye decolorization [74, 75]. Our findings revealed amylase production by six halophilic strains, namely H. meridiana, S. costicola, H. karajensis, S. salis, P. shioyasakiensis, and S. amylolyticus. On the other hand, V. oceani, V. marismortui, M. lipolyticus, and P. salipaludis showed no amylase activity. In good agreement, bacterial strains isolated from saline habitats showed significant amylase activity, such as Halobacillus sp. [76], and Pseudoalteromonas sp. M175 [77]. About 58 strains of Vibrio spp. isolated from marine habitats in Malaysia exhibited amylase activity [15]. Furthermore, amylase production by numerous microorganisms isolated from saline and hypersaline habitats has been well documented, such as Bacillus subtilis S8-1 [78], Streptomyces sp. D1 [79], Vibrio sp. [80], Wangia sp. C52 [81], Pontibacillus chungwhensis [82], Bacillus barbaricus [82], Nocardiopsis sp. B2 [83], and Anoxybacillus beppuensis TSSC-1 [84]. In this study, we identified six protease-producing halophilic strains, V. oceani, V. marismortui, H. karajensis, S. salis, P. shioyasakiensis, and P. salipaludis. Similarly, previous studies have reported the production of proteases from bacterial strains isolated from habitats with high salinity, such as Pseudoalteromonas sp. strain CP76 [85], Halobacillus karajensis [86], Virgibacillus marismortui [87], Virgibacillus sp. EMB13 [88], Halobacillus sp. CJ4 [89], Virgibacillus sp. SK33 [90], Halobacillus sp. LY6 [91], Salicola sp. IC10 [92], Halobacillus sp. SCSIO 20089 [93], as well as Halobacillus andaensis, H. dabanensis and Salicola marasensis [94]. Similarly, various halophilic and halotolerant bacteria have been reported as significant producers of protease such as Halococcus agarilyticus [37], Bacillus sp. [95], Bacillus firmus CAS 7 [96], and Pseudomonas SD11 [97]. Our findings harmony with those of previous studies that reported the absence of extracellular protease activity in various species belonging to the genus Salinivibrio, such as Salinivibrio costicola NCIMB 13595 and Salinicoccus roseus DSM 5351 [98].

In this investigation, we identified seven promising lipase-producing halophilic bacterial strains: H. meridiana, S. costicola, V. oceani, M. lipolyticus, H. karajensis, S. salis, and P. salipaludis. Whereas V. marismortui, P. shioyasakiensis, and S. amylolyticus exhibited no lipase activity. Likewise, various bacterial strains recovered from saline or hypersaline habitats displayed significant lipase production, such as Marinobacter lipolytica [45], Marinobacter sp. EMB5 [99], Marinobacter adhaerens t76_800 [100], Haloarcula sp. G41 [101], Aeromonas sp. EBB-1 [102], Vibrio sp. [103], Vibrio vulnificus [104], Salinivibrio costicola DSM 8285 [98], Salinivibrio costicola NCIMB 13595 [98], Halobacillus sp. strain AR11 [49], Halobacillus trueperi RSK CAS9 [105], Virgibacillus alimentarius [106], Salicola sp. IC10 [92], and Salicola marasensis [94]. Hydrolysis by β-glucosidase is considered a significant step in the hydrolysis of lignocellulosic materials [107] by cellulases that hydrolyze cellulose into simple units by cleaving β-1,4-glycosidic linkages [108, 109]. These enzymes are classified into three types; exoglucanases, endoglucanases, and β-glucosidases [110]. For β-glucosidase, we identified five promising producers: V. oceani, V. marismortui, H. karajensis, P. shioyasakiensis and S. amylolyticus. However, H. meridiana, S. costicola, M. lipolyticus, S. salis, and P. salipaludis did not exhibit extracellular β-glucosidase activity. In this connection, Pseudoalteromonas sp. GXQ-1 showed significant activity of β-glucosidase [55]. Moreover, many marine-derived bacteria have been reported as a significant source for the production of β-glucosidase such as Streptomyces sp.

In conclusion, this investigation shed light on the diversity of halophilic bacterial strains from the extreme ecosystem, Mediterranean solar saltern in Egypt. The majority of the recovered halophilic strains were able to produce salt-tolerant hydrolytic enzymes. We found that amylase was the most prevalent enzyme, while lipase was uncommon. Furthermore, ten extreme halophilic bacterial strains belonging to nine genera were identified. Of these strains, H. meridiana SHB1, V. marismortui SHB21, and S. amylolyticus SHB59 showed remarkable growth in the presence of 25% NaCl and produced at least two halotolerant extracellular enzymes. Nonetheless, future studies are planned to purify and characterize these enzymes to exploit their merits in various industrial applications.

Conflict of Interest

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The authors have no financial conflicts of interest to declare.

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