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Antimicrobial protein candidates from the thermophilic Geobacillus sp. Strain ZGt-1 : Production, proteomics, and bioinformatics analysis

Alkhalili, Rawana N. LU ; Bernfur, Katja LU ; Dishisha, Tarek LU ; Mamo, Gashaw LU ; Schelin, Jenny LU ; Canbäck, Björn LU ; Emanuelsson, Cecilia LU and Hatti-Kaul, Rajni LU (2016) In International Journal of Molecular Sciences 17(8).
Abstract

A thermophilic bacterial strain, Geobacillus sp. ZGt-1, isolated from Zara hot spring in Jordan, was capable of inhibiting the growth of the thermophilic G. stearothermophilus and the mesophilic Bacillus subtilis and Salmonella typhimurium on a solid cultivation medium. Antibacterial activity was not observed when ZGt-1 was cultivated in a liquid medium; however, immobilization of the cells in agar beads that were subjected to sequential batch cultivation in the liquid medium at 60 °C showed increasing antibacterial activity up to 14 cycles. The antibacterial activity was lost on protease treatment of the culture supernatant. Concentration of the protein fraction by ammonium sulphate precipitation followed by denaturing polyacrylamide... (More)

A thermophilic bacterial strain, Geobacillus sp. ZGt-1, isolated from Zara hot spring in Jordan, was capable of inhibiting the growth of the thermophilic G. stearothermophilus and the mesophilic Bacillus subtilis and Salmonella typhimurium on a solid cultivation medium. Antibacterial activity was not observed when ZGt-1 was cultivated in a liquid medium; however, immobilization of the cells in agar beads that were subjected to sequential batch cultivation in the liquid medium at 60 °C showed increasing antibacterial activity up to 14 cycles. The antibacterial activity was lost on protease treatment of the culture supernatant. Concentration of the protein fraction by ammonium sulphate precipitation followed by denaturing polyacrylamide gel electrophoresis separation and analysis of the gel for antibacterial activity against G. stearothermophilus showed a distinct inhibition zone in 15-20 kDa range, suggesting that the active molecule(s) are resistant to denaturation by SDS. Mass spectrometric analysis of the protein bands around the active region resulted in identification of 22 proteins with molecular weight in the range of interest, three of which were new and are here proposed as potential antimicrobial protein candidates by in silico analysis of their amino acid sequences. Mass spectrometric analysis also indicated the presence of partial sequences of antimicrobial enzymes, amidase, and DD-carboxypeptidase.

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organization
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type
Contribution to journal
publication status
published
subject
keywords
Antimicrobial proteins, Cell-recycling, Food spoilage bacteria, Geobacillus, Immobilization, SDS-resistant proteins, Thermophile
in
International Journal of Molecular Sciences
volume
17
issue
8
publisher
MOLECULAR DIVERSITY PRESERVATION INT
external identifiers
  • scopus:84983466657
  • wos:000382337900172
ISSN
1661-6596
DOI
10.3390/ijms17081363
language
English
LU publication?
yes
id
17571671-0f82-4e29-8144-ad890d20bf4b
date added to LUP
2016-12-02 14:41:12
date last changed
2017-01-01 08:41:37
@article{17571671-0f82-4e29-8144-ad890d20bf4b,
  abstract     = {<p>A thermophilic bacterial strain, Geobacillus sp. ZGt-1, isolated from Zara hot spring in Jordan, was capable of inhibiting the growth of the thermophilic G. stearothermophilus and the mesophilic Bacillus subtilis and Salmonella typhimurium on a solid cultivation medium. Antibacterial activity was not observed when ZGt-1 was cultivated in a liquid medium; however, immobilization of the cells in agar beads that were subjected to sequential batch cultivation in the liquid medium at 60 °C showed increasing antibacterial activity up to 14 cycles. The antibacterial activity was lost on protease treatment of the culture supernatant. Concentration of the protein fraction by ammonium sulphate precipitation followed by denaturing polyacrylamide gel electrophoresis separation and analysis of the gel for antibacterial activity against G. stearothermophilus showed a distinct inhibition zone in 15-20 kDa range, suggesting that the active molecule(s) are resistant to denaturation by SDS. Mass spectrometric analysis of the protein bands around the active region resulted in identification of 22 proteins with molecular weight in the range of interest, three of which were new and are here proposed as potential antimicrobial protein candidates by in silico analysis of their amino acid sequences. Mass spectrometric analysis also indicated the presence of partial sequences of antimicrobial enzymes, amidase, and DD-carboxypeptidase.</p>},
  articleno    = {1363},
  author       = {Alkhalili, Rawana N. and Bernfur, Katja and Dishisha, Tarek and Mamo, Gashaw and Schelin, Jenny and Canbäck, Björn and Emanuelsson, Cecilia and Hatti-Kaul, Rajni},
  issn         = {1661-6596},
  keyword      = {Antimicrobial proteins,Cell-recycling,Food spoilage bacteria,Geobacillus,Immobilization,SDS-resistant proteins,Thermophile},
  language     = {eng},
  month        = {08},
  number       = {8},
  publisher    = {MOLECULAR DIVERSITY PRESERVATION INT},
  series       = {International Journal of Molecular Sciences},
  title        = {Antimicrobial protein candidates from the thermophilic Geobacillus sp. Strain ZGt-1 : Production, proteomics, and bioinformatics analysis},
  url          = {http://dx.doi.org/10.3390/ijms17081363},
  volume       = {17},
  year         = {2016},
}