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

Extremophiles. 2015 Jul;19(4):763-74. doi: 10.1007/s00792-015-0752-3. Epub 2015 May 16.

Abstract

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.

MeSH terms

  • Archaeal Proteins* / chemistry
  • Archaeal Proteins* / isolation & purification
  • Egypt
  • Halobacterium* / enzymology
  • Halobacterium* / isolation & purification
  • Lakes / microbiology*
  • Peptide Hydrolases* / chemistry
  • Peptide Hydrolases* / isolation & purification

Substances

  • Archaeal Proteins
  • Peptide Hydrolases