Isolation and identification of alkaline protease producer halotolerantBacillus licheniformis strain BA17

Ateş, Özlem; Öner, Ebru Toksoy; Arikan, Burhan; Denizci, Aziz Akın; Kazan, Dilek

doi:10.1007/bf03175075

Cited by 6 publications

(8 citation statements)

References 47 publications

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“…The enzyme production was also high in the production medium containing lactose (2.3 U/ml) as the carbon source and ammonium chloride as the nitrogen source (2.3 U/ml). Glucose (1.0%) repressed enzyme secretion which was similar to the results reported in Bacillus licheniformis (Ateş et al 2007). In the case of halophilic bacterium Chromohalobacter sp.…”

Section: Discussionsupporting

confidence: 86%

Biosynthesis optimization and purification of a solvent stable alkaline serine protease from Halobacterium sp.

Vijayaraghavan

Jebamalar²,

Vincent

2011

Ann Microbiol

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Three halotolerant bacterial isolates that were isolated from the salt pan of coastal Tamilnadu, India, were screened on skimmed milk agar plates containing 1-25% (w/v) NaCl for protease secretion. Among the three positive isolates, Halobacterium sp. showed the maximum protease secretion (4.5 mm diameter zone) and was hence selected for this study. The enzyme was a serine protease as evidenced by complete inactivation of enzyme after treatment with a serine protease inhibitor (phenylmethylsulfonyl fluoride). Culture conditions which optimize protease production that included pH, fermentation period, NaCl in addition to different factors such as carbon and nitrogen sources, casein concentration were investigated. Optimum bacterial growth and proteolytic activity was achieved after 48 h of incubation at 37°C, pH 10.0 with 5% (w/v) NaCl concentration and 2.5% (w/v) casein. Initial experiments revealed that the culture medium containing 1.0% (w/v) lactose as the carbon source and 1.0% (w/v) ammonium chloride as the nitrogen source supported maximized enzyme production. Analysis carried out using SDS-PAGE revealed that the extracellular enzyme consisted of a single polypeptide chain of 43 kDa which was purified by a combination of ammonium sulphate fractionation, DEAE cellulose and Sephadex G-75 gel filtration chromatography. This protease was stable for a wide range of temperatures (40-60°C), NaCl concentrations (1-5%, w/v) and pH (8.0-10.0) with maximal activity observed at 60°C, 5% NaCl and pH 10.0. Proteolytic activity was enhanced by 1.0 mM of Ca 2+ (137%) and strongly inhibited by Mn 2+ (67%). Most importantly, the proteolytic activity remained stable in alkaline pH and was found to be active at higher temperatures. The broad range of temperature optima with respect to enzyme activity was indicative of this enzymes suitability in fish sauce fermentation, which was carried out at 35-45°C. This enzyme was stable in organic solvents such as toluene and hexane and the relative enzyme activity was 103 and 114%, respectively, which mainly finds use in the paint industry.

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Section: Discussionsupporting

confidence: 86%

Biosynthesis optimization and purification of a solvent stable alkaline serine protease from Halobacterium sp.

Vijayaraghavan

Jebamalar²,

Vincent

2011

Ann Microbiol

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“…Bacillus licheniformis BA17 was isolated and identified on the basis of 16S rRNA gene sequencing as described previously (Ates et al, 2007). 16S rDNA sequence of the strain BA17 has been deposited in the NCBI databases under the accession number DQ176435.…”

Section: Microorganism and Protease Production Halotolerantmentioning

confidence: 99%

“…16S rDNA sequence of the strain BA17 has been deposited in the NCBI databases under the accession number DQ176435. Alkaline protease production from B. licheniformis BA17 was carried out in the medium containing (% w/v) 1.0 maltose, 0.5 yeast extract, 0.1 K 2 HPO 4 , 1 Na 2 NO 3 and 0.02 MgSO 4 ·7H 2 O at 37 °C and 180 rpm (Ates et al, 2007). Medium pH was adjusted to 10.0 by aseptic addition of 10% Na 2 CO 3 solution after sterilization.…”

Section: Microorganism and Protease Production Halotolerantmentioning

confidence: 99%

Alkaline serine protease from halotolerantBacillus licheniformis BA17

Öztürk¹,

Özeren-Morgan²,

Dilgimen³

et al. 2009

Ann. Microbiol.

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An alkaline protease from halotolerant Bacillus licheniformis BA17, isolated from Van Lake in Turkey, was purified 5.4 fold with 58% yield. The molecular weight was 19.7 kDa and the optimum temperature and pH were 60 °C and 10, respectively. The halflife of the pure enzyme was 38 h, 93 min, 14 min and 6 min at 40, 50, 60 and 70 °C, respectively. BA17 protease is very active at 30 °C between pH 8.0 and 10. Enzyme activity increased in the presence of Cu +2 , Mg +2 , Mn +2 and K +1 ions. Enzyme retained activity with 5% SDS (w/v) and 1% Triton X-100 (v/v). Inhibition with PMSF and EDTA suggested that the enzyme is a serine protease and is a metal-activated enzyme. Based on the N-terminal sequence of the first 13 amino acids, B. licheniformis BA17 alkaline protease did not show identity to any of those from other Bacillus species.

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“…Those from Bacillus species have wide use and importance in several industrial economies such as in food, detergent and in the synthesis of biologically active peptides 4 . Extremozymes have a great biotechnological potential, including agricultural, chemical and pharmaceutical applications 5,6 .…”

Section: Quick Response Codementioning

confidence: 99%

“…The bacteria survive in the soda lakes are mostly alkaliphilic, it is to be considered that extracellular enzymes synthesized by alkaliphilic bacteria would be diligent under alkaline stipulations. These extremozymes are active in absence or presence of trace amount of some metal ions and such enzymes are of biotechnological potential 6 . The Lonar lake is the great soda lake which is located in Lonar, District Buldhana, Maharashtra, India (latitude 19° 8, longitude 76° 36).…”

Section: Quick Response Codementioning

confidence: 99%

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2018

IJPSR

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Culture dependent phenotypic characterization analyses were applied to study the protease producing bacteria from Indian Soda (Lonar crater). The uniqueness of the Lonar lake water is its salinity and alkalinity. A total of one hundred and fourteen bacterial cultures were isolated from the water and sediment samples collected from the hyper alkaline saline environment of Lonar lake. The thirty two bacterial strains were found protease producing bacterial culture, out of them strain EW2 was selected for 16S rDNA sequencing, production and partial characterizations of protease on the basis of their maximum proteolytic activity. The phylogenetic position indicated the strain was related to phylum Firmicutes. Optimum enzyme activity was found to be at 80 °C and pH 9.5. The protease was highly stable over broad temperature from 40 to 90 °C, pH 7.0-12.0, showing excellent thermostablity and alkaline tolerant nature. The enzyme activity has enhanced by all metal ion which was tested in this studies indicating it was a metalloenzyme but inhibited by organic solvent except acetone. This is valuable information for enzyme production and optimization of protease from Bacillus pumilus EW2 has bright future towards the improvement and production of novel enzymes for leather, detergent and pharmaceuticals industries and biotechnological potential.

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Isolation and identification of alkaline protease producer halotolerantBacillus licheniformis strain BA17

Cited by 6 publications

References 47 publications

Biosynthesis optimization and purification of a solvent stable alkaline serine protease from Halobacterium sp.

Biosynthesis optimization and purification of a solvent stable alkaline serine protease from Halobacterium sp.

Alkaline serine protease from halotolerantBacillus licheniformis BA17

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