PARTIAL PURIFICATION AND CHARACTERIZATION OF AN EXTRACELLULAR METALLOPEPTIDASE PRODUCED BY Bacillus amyloliquefaciens FE-K1
The aim of this study was to purify and characterize the peptidase of Bacillus amyloliquefaciens (Fukumoto) (strain FE-K1) isolated from ropey bread. Peptidases were purified from crude enzyme solution by affinity chromatography with an efficiency of 25 % and a purification coefficient of 1.53. The...
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| Format: | Article |
| Language: | English |
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Trakya University
2020-04-01
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| Series: | Trakya University Journal of Natural Sciences |
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| Online Access: | https://dergipark.org.tr/en/download/article-file/1032880 |
| _version_ | 1797633903314337792 |
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| author | Fundagül Erem Mehmet İnan Barçın Karakaş Budak Muharrem Certel |
| author_facet | Fundagül Erem Mehmet İnan Barçın Karakaş Budak Muharrem Certel |
| author_sort | Fundagül Erem |
| collection | DOAJ |
| description | The aim of this study was to purify and characterize the peptidase of Bacillus amyloliquefaciens (Fukumoto) (strain FE-K1) isolated from ropey bread. Peptidases were purified from crude enzyme solution by affinity chromatography with an efficiency of 25 % and a purification coefficient of 1.53. The optimum pH of partially purified peptidase (PPPase) solution was determined as 7.5 and the peptidases retained approximately 90 % of their initial activity in the pH range 7.0-8.5 following incubation at 37°C for 2 h. The optimum temperature for the PPPase was 60°C. The approximate molecular weight of the PPPase was determined as 36 kDa. Inactivation of the PPPase in the presence of O-FEN and EDTA showed them to be metallopeptidases and 5 mM of K+1 and 5 mM of Mn+2 ions increased the enzyme activity by 4 % and 6.15 %, respectively. The presence of Hg+2, Fe+3 and SDS (0.1-1.0 % w/v) caused inactivation whereas the enzyme retained most of its activity in the presence of 0.1-1.0 % (v/v) Triton X-100, Tween 20 and Tween 80 and 1-20 % (v/v) xylene, ethanol, acetone and acetonitrile. Characterization of the PPPase revealed the enzyme as a neutral serine metallopeptidase compatible with some organic solvents and surfactants. |
| first_indexed | 2024-03-11T12:00:11Z |
| format | Article |
| id | doaj.art-e9104fad2b7046d7aa992c18e15c34c5 |
| institution | Directory Open Access Journal |
| issn | 2528-9691 |
| language | English |
| last_indexed | 2024-03-11T12:00:11Z |
| publishDate | 2020-04-01 |
| publisher | Trakya University |
| record_format | Article |
| series | Trakya University Journal of Natural Sciences |
| spelling | doaj.art-e9104fad2b7046d7aa992c18e15c34c52023-11-08T07:27:56ZengTrakya UniversityTrakya University Journal of Natural Sciences2528-96912020-04-01211476110.23902/trkjnat.64752569PARTIAL PURIFICATION AND CHARACTERIZATION OF AN EXTRACELLULAR METALLOPEPTIDASE PRODUCED BY Bacillus amyloliquefaciens FE-K1Fundagül Erem0Mehmet İnan1Barçın Karakaş Budak2Muharrem Certel3ZONGULDAK BÜLENT ECEVİT ÜNİVERSİTESİAKDENIZ UNIVERSITYAKDENIZ UNIVERSITYAKDENİZ ÜNİVERSİTESİThe aim of this study was to purify and characterize the peptidase of Bacillus amyloliquefaciens (Fukumoto) (strain FE-K1) isolated from ropey bread. Peptidases were purified from crude enzyme solution by affinity chromatography with an efficiency of 25 % and a purification coefficient of 1.53. The optimum pH of partially purified peptidase (PPPase) solution was determined as 7.5 and the peptidases retained approximately 90 % of their initial activity in the pH range 7.0-8.5 following incubation at 37°C for 2 h. The optimum temperature for the PPPase was 60°C. The approximate molecular weight of the PPPase was determined as 36 kDa. Inactivation of the PPPase in the presence of O-FEN and EDTA showed them to be metallopeptidases and 5 mM of K+1 and 5 mM of Mn+2 ions increased the enzyme activity by 4 % and 6.15 %, respectively. The presence of Hg+2, Fe+3 and SDS (0.1-1.0 % w/v) caused inactivation whereas the enzyme retained most of its activity in the presence of 0.1-1.0 % (v/v) Triton X-100, Tween 20 and Tween 80 and 1-20 % (v/v) xylene, ethanol, acetone and acetonitrile. Characterization of the PPPase revealed the enzyme as a neutral serine metallopeptidase compatible with some organic solvents and surfactants.https://dergipark.org.tr/en/download/article-file/1032880bacillus amyloliquefaciensmetallopeptidaseropy breadpurification |
| spellingShingle | Fundagül Erem Mehmet İnan Barçın Karakaş Budak Muharrem Certel PARTIAL PURIFICATION AND CHARACTERIZATION OF AN EXTRACELLULAR METALLOPEPTIDASE PRODUCED BY Bacillus amyloliquefaciens FE-K1 Trakya University Journal of Natural Sciences bacillus amyloliquefaciens metallopeptidase ropy bread purification |
| title | PARTIAL PURIFICATION AND CHARACTERIZATION OF AN EXTRACELLULAR METALLOPEPTIDASE PRODUCED BY Bacillus amyloliquefaciens FE-K1 |
| title_full | PARTIAL PURIFICATION AND CHARACTERIZATION OF AN EXTRACELLULAR METALLOPEPTIDASE PRODUCED BY Bacillus amyloliquefaciens FE-K1 |
| title_fullStr | PARTIAL PURIFICATION AND CHARACTERIZATION OF AN EXTRACELLULAR METALLOPEPTIDASE PRODUCED BY Bacillus amyloliquefaciens FE-K1 |
| title_full_unstemmed | PARTIAL PURIFICATION AND CHARACTERIZATION OF AN EXTRACELLULAR METALLOPEPTIDASE PRODUCED BY Bacillus amyloliquefaciens FE-K1 |
| title_short | PARTIAL PURIFICATION AND CHARACTERIZATION OF AN EXTRACELLULAR METALLOPEPTIDASE PRODUCED BY Bacillus amyloliquefaciens FE-K1 |
| title_sort | partial purification and characterization of an extracellular metallopeptidase produced by bacillus amyloliquefaciens fe k1 |
| topic | bacillus amyloliquefaciens metallopeptidase ropy bread purification |
| url | https://dergipark.org.tr/en/download/article-file/1032880 |
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