Purification and characterization of an extreme halothermophilic protease from a halophilic bacterium Chromohalobacter sp. TVSP101

Vidyasagar, M.; Prakash, S.; Mahajan, Vineet; Shouche, Yogesh S.; Sreeramulu, K.

doi:10.1590/s1517-83822009000100002

Cited by 43 publications

(22 citation statements)

References 28 publications

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“…Asker et al [23] purified two proteases, namely P1 and P2 from the bacterial strain Bacillus megaterium, and determined their molecular weight as 28 and 25 KDa, respectively. Vidyasagar et al [25] purified an extreme thermophilic protease (66 kDa) from Chromohalobacter sp. TVSP101.…”

Section: Characterization Of Purified Protease and Amylasementioning

confidence: 99%

Purification and Characterization of Extracellular Protease and Amylase Produced by the Bacterial Strain, Corynebacterium alkanolyticum ATH3 Isolated from Fish Gut

et al. 2015

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The use of enzymes in different industrial sectors increased significantly due to huge industrialization. Different types of protease and amylase are randomly used in industries including food, textile, and paper. For this purpose, purification of extracellular protease and amylase produced by the bacterium, Corynebacterium alkanolyticum ATH3 (Acc. No. JX656749) isolated from the distal intestine of a freshwater fish, Anabas testudineus, was carried out using column chromatography. The specific activity of protease and amylase significantly increased with each step of purification and finally became 93.73 and 88.1 U/mg protein with a purification fold 26.03 and 44.94, respectively. The sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis showed molecular weight of purified protease and amylase was ∼17 and ∼28 kDa, respectively. To the authors' knowledge, this is the first report of ∼17 kDa protease and ∼28 kDa amylase from the bacterial strain C. alkanolyticum ATH3. Further, enzyme activity was also evidenced by zymography analysis. The enzymes acted optimally at pH 7.5-8.0 and temperature 35-45 • C, respectively. So, due to cheapest source, these two enzymes are very important for various purposes in industrial sectors.Keywords Fish gut bacteria · Protease and amylase purification · SDS-PAGE · Zymography analysis · pH and temperature sensitivity B Arun K. Ray

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Section: Characterization Of Purified Protease and Amylasementioning

confidence: 99%

Purification and Characterization of Extracellular Protease and Amylase Produced by the Bacterial Strain, Corynebacterium alkanolyticum ATH3 Isolated from Fish Gut

et al. 2015

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“…PB407 (Kanlayakrit et al 2005), 60°C for proteinase from Natronococcus occultus (Studdert et al 2001) and 75°C for proteinase from Chromohalobacter sp. TVSP101 (Vidyasagar et al 2009). The proteinase was highly active over a broad pH range with maximum activity at pH 10.0, indicating its alkalitolerant nature.…”

Section: Discussionmentioning

confidence: 99%

Production and characterization of a novel extracellular metalloproteinase by a newly isolated moderate halophile, Halobacillus sp. LY6

Liu

et al. 2011

Folia Microbiol

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A moderately halophilic bacterium LY6 with high proteolytic activity was isolated. Biochemical and physiological characterization, along with 16S rDNA sequence analysis placed the isolate in the genus Halobacillus. The salinity of the culture medium strongly influenced the proteinase production of LY6. Maximum enzyme production was observed in the medium containing 5% Na(2)SO(4) or 10% NaCl. Proteinase production was synchronized with bacterial growth and reached a maximum level during the mid-stationary phase. Enzyme purification was carried out by a simple approach including a combination of ammonium sulfate precipitation and Sephacryl S-100 gel filtration chromatography. SDS-PAGE and gelatin zymography analysis revealed it was a monomer with high molecular weight of 69 kDa. Optimal proteinase activity was obtained at pH 10.0, 40°C, and 10% NaCl. It was high active over broad temperature (30-80°C), pH (6.0-12.0), and NaCl concentration (0-25%) ranges, indicating its thermostable, alkali-stable, and halotolerant nature. Moreover, the enzyme activity was markedly enhanced by Ca(2+) and Cu(2+), but strongly inhibited by EDTA, PAO, and DEPC, indicating that it probably was a metalloproteinase with cysteine and histidine residues located in its active site.

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“…Inhibitor studies on the halophilic protease of strain HP25 were performed according to the protocol described by Vidyasagar et al (2009). The assay mixture (1 mL) was prepared by combining 500 µL of the purified halophilic protease solution (pH 8.0), followed by adding 500 µL of each inhibitor solution ( Table 2).…”

Section: Effect Of Inhibitors On the Halophilic Protease Activitymentioning

confidence: 99%

Purification and characterization of halo-alkali-thermophilic protease from Halobacterium sp. strain HP25 isolated from raw salt, Lake Qarun, Fayoum, Egypt

2015

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A total of 33 halophilic protease producers were isolated from different salt samples collected from Emisal salt company at Lake Qarun, Fayoum, Egypt. Of these strains, an extremely halophilic strain that grew optimally at 30 % (w/v) NaCl was characterized and identified as Halobacterium sp. strain HP25 based on 16S rRNA gene sequencing and phenotypic characterization. A halo-alkali-thermophilic protease was purified in three successive steps from the culture supernatant. The purified halophilic protease consisted of a single polypeptide chain with a molecular mass of 21 kDa and was enriched 167-fold to a specific activity of 6350 U mg(-1). The purified enzyme was active over a broad pH range from 6.0 to 11.0, with maximum activity at pH 8.0, exhibited a broad temperature range from 30 to 80 °C with optimum activity at 60 °C, and was active at salt concentrations ranging from 5 to 25 % (w/v), with optimum activity at 17 % NaCl (w/v). The K M and V max values of the purified halophilic protease with casein as a substrate were 523 µg mL(-1) and 2500 µg min(-1) mL(-1), respectively. In addition, this enzyme was stable in the tested organic solvents and laundry detergents such methanol, propanol, butanol, hexane, Persil and Ariel. The unusual properties of this enzyme allow it to be used for various applications, such as the ripening of salted fish. Furthermore, its stability and activity in the presence of organic solvents and detergents also allow the use of this enzyme for further novel applications and as an additive in detergent formulations.

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Purification and characterization of an extreme halothermophilic protease from a halophilic bacterium Chromohalobacter sp. TVSP101

Cited by 43 publications

References 28 publications

Purification and Characterization of Extracellular Protease and Amylase Produced by the Bacterial Strain, Corynebacterium alkanolyticum ATH3 Isolated from Fish Gut

Purification and Characterization of Extracellular Protease and Amylase Produced by the Bacterial Strain, Corynebacterium alkanolyticum ATH3 Isolated from Fish Gut

Production and characterization of a novel extracellular metalloproteinase by a newly isolated moderate halophile, Halobacillus sp. LY6

Purification and characterization of halo-alkali-thermophilic protease from Halobacterium sp. strain HP25 isolated from raw salt, Lake Qarun, Fayoum, Egypt

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