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Novel alkaline proteases from alkaliphilic bacteria grown using chicken feather.

Gessesse, A; Hatti-Kaul, Rajni LU ; Gashe, B and Mattiasson, Bo LU (2003) In Enzyme and Microbial Technology 32(5). p.519-524
Abstract
Two alkaline protease producing alkaliphilic bacterial strains, designated as AL-20 and AL-89, were isolated from a naturally occurring alkaline habitat. The two strains were identified as Nesternkonia sp. and Bacillus pseudofirmus, respectively. Both strains grew and produced alkaline protease using feather as the sole source of carbon and nitrogen. Addition of 0.5% glucose to the feather medium increased protease production by B. pseudofirmus AL-89 and suppressed enzyme production by Nesternkonia sp. AL-20. The enzymes from both organisms were purified to electrophoretic homogeneity following ammonium sulphate precipitation, ion exchange, hydrophobic interaction, and gel filtration chromatography. The molecular weight, determined using... (More)
Two alkaline protease producing alkaliphilic bacterial strains, designated as AL-20 and AL-89, were isolated from a naturally occurring alkaline habitat. The two strains were identified as Nesternkonia sp. and Bacillus pseudofirmus, respectively. Both strains grew and produced alkaline protease using feather as the sole source of carbon and nitrogen. Addition of 0.5% glucose to the feather medium increased protease production by B. pseudofirmus AL-89 and suppressed enzyme production by Nesternkonia sp. AL-20. The enzymes from both organisms were purified to electrophoretic homogeneity following ammonium sulphate precipitation, ion exchange, hydrophobic interaction, and gel filtration chromatography. The molecular weight, determined using SDS–PAGE, was 23 kDa for protease AL-20 and 24 kDa for protease AL-89. Protease AL-20 was active in a broad pH range displaying over 90% of its maximum activity between pH 7.5 and 11.5 with a peak at pH 10. The enzyme is unique in that unlike all other microbial serine proteases known so far, it did not require Ca2+ for activity and thermal stability. Its optimum temperature for activity was at 70 °C and was stable after 1 h incubation at 65 °C both in the presence and absence of Ca2+. These properties make protease AL-20 an ideal candidate for detergent application. Protease AL-89 on the other hand require Ca2+ for activity and stability at temperature values above 50 °C. Its optimum activity was at 60 and 70 °C in the absence and presence of Ca2+, respectively. It displayed a pH optimum of 11 and retained about 70% or more of its original activity between pH 6.5 and 11. B. pseudofirmus AL-89, and the protease it produce offers an interesting potential for the enzymatic and/or microbiological hydrolysis of feather to be used as animal feed supplement. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Alkaline protease, Feather degradation, Alkaliphile, Bacillus, Nesternkonia, Calcium independent protease, Keratinase, Soda lake
in
Enzyme and Microbial Technology
volume
32
issue
5
pages
519 - 524
publisher
Elsevier
external identifiers
  • wos:000181908500003
  • scopus:0037426295
ISSN
0141-0229
DOI
10.1016/S0141-0229(02)00324-1
language
English
LU publication?
yes
id
73651f8e-4a5b-4fab-8375-d646869983b4 (old id 129130)
date added to LUP
2007-07-02 10:36:51
date last changed
2018-10-21 03:39:39
@article{73651f8e-4a5b-4fab-8375-d646869983b4,
  abstract     = {Two alkaline protease producing alkaliphilic bacterial strains, designated as AL-20 and AL-89, were isolated from a naturally occurring alkaline habitat. The two strains were identified as Nesternkonia sp. and Bacillus pseudofirmus, respectively. Both strains grew and produced alkaline protease using feather as the sole source of carbon and nitrogen. Addition of 0.5% glucose to the feather medium increased protease production by B. pseudofirmus AL-89 and suppressed enzyme production by Nesternkonia sp. AL-20. The enzymes from both organisms were purified to electrophoretic homogeneity following ammonium sulphate precipitation, ion exchange, hydrophobic interaction, and gel filtration chromatography. The molecular weight, determined using SDS–PAGE, was 23 kDa for protease AL-20 and 24 kDa for protease AL-89. Protease AL-20 was active in a broad pH range displaying over 90% of its maximum activity between pH 7.5 and 11.5 with a peak at pH 10. The enzyme is unique in that unlike all other microbial serine proteases known so far, it did not require Ca2+ for activity and thermal stability. Its optimum temperature for activity was at 70 °C and was stable after 1 h incubation at 65 °C both in the presence and absence of Ca2+. These properties make protease AL-20 an ideal candidate for detergent application. Protease AL-89 on the other hand require Ca2+ for activity and stability at temperature values above 50 °C. Its optimum activity was at 60 and 70 °C in the absence and presence of Ca2+, respectively. It displayed a pH optimum of 11 and retained about 70% or more of its original activity between pH 6.5 and 11. B. pseudofirmus AL-89, and the protease it produce offers an interesting potential for the enzymatic and/or microbiological hydrolysis of feather to be used as animal feed supplement.},
  author       = {Gessesse, A and Hatti-Kaul, Rajni and Gashe, B and Mattiasson, Bo},
  issn         = {0141-0229},
  keyword      = {Alkaline protease,Feather degradation,Alkaliphile,Bacillus,Nesternkonia,Calcium independent protease,Keratinase,Soda lake},
  language     = {eng},
  number       = {5},
  pages        = {519--524},
  publisher    = {Elsevier},
  series       = {Enzyme and Microbial Technology},
  title        = {Novel alkaline proteases from alkaliphilic bacteria grown using chicken feather.},
  url          = {http://dx.doi.org/10.1016/S0141-0229(02)00324-1},
  volume       = {32},
  year         = {2003},
}