Isolation and characterization of Bacillus sp. IMV B-7883 proteases
The representatives of Bacillus are some of the best protease producers studied so far since they exhibit broad substrate specificity, significant activity, stability, simple downstream purification, short period of fermentation and low cost. Earlier, we showed that Bacillus sp. IMV B-7883 strain sy...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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National Academy of Sciences of Ukraine, Palladin Institute of Biochemistry
2023-10-01
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| Series: | The Ukrainian Biochemical Journal |
| Subjects: | |
| Online Access: | http://ukrbiochemjournal.org/wp-content/uploads/2023/11/Gudzenko_95_5.pdf |
| _version_ | 1797361914741784576 |
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| author | O. V. Gudzenko L. D. Varbanets |
| author_facet | O. V. Gudzenko L. D. Varbanets |
| author_sort | O. V. Gudzenko |
| collection | DOAJ |
| description | The representatives of Bacillus are some of the best protease producers studied so far since they exhibit broad substrate specificity, significant activity, stability, simple downstream purification, short period of fermentation and low cost. Earlier, we showed that Bacillus sp. IMV B-7883 strain synthesizes an extracellular proteases, which exhibit elastolytic and fibrinogenolytic activity. The aim of the work was to isolate and purify these enzymes from the culture liquid of the Bacillus sp. IMV B-7883 strain, as well as to study their properties. Isolation and purification of proteases was carried out by precipitation of the culture liquid with ammonium sulfate, gel permeation and ion exchange chromatography and rechromatography on Sepharose 6B. As a result, proteases with elastolytic and fibrinogenolytic activity with a molecular weight of 23 and 20 kDa respectively were isolated with elastase activity increased by 63.6 and fibrinogenolytic activity by 44.1 times. The enzyme with elastase activity had a pH-optimum of 7.0 and hydrolyzed only elastin, while the enzyme with fibrinogenolytic activity was an alkaline protease with a pH-optimum of 8.0 and in addition to fibrinogen, showed specificity for fibrin and, in trace amounts, for collagen. |
| first_indexed | 2024-03-08T15:59:58Z |
| format | Article |
| id | doaj.art-e60297ee99924e75b1d6d0db1e6afd40 |
| institution | Directory Open Access Journal |
| issn | 2409-4943 2413-5003 |
| language | English |
| last_indexed | 2024-03-08T15:59:58Z |
| publishDate | 2023-10-01 |
| publisher | National Academy of Sciences of Ukraine, Palladin Institute of Biochemistry |
| record_format | Article |
| series | The Ukrainian Biochemical Journal |
| spelling | doaj.art-e60297ee99924e75b1d6d0db1e6afd402024-01-08T10:40:49ZengNational Academy of Sciences of Ukraine, Palladin Institute of BiochemistryThe Ukrainian Biochemical Journal2409-49432413-50032023-10-019559810710.15407/ubj95.05.098Isolation and characterization of Bacillus sp. IMV B-7883 proteasesO. V. Gudzenko0L. D. Varbanets1Zabolotny Institute of Microbiology and Virology, National Academy of Sciences of Ukraine, Kyiv, UkraineZabolotny Institute of Microbiology and Virology, National Academy of Sciences of Ukraine, Kyiv, UkraineThe representatives of Bacillus are some of the best protease producers studied so far since they exhibit broad substrate specificity, significant activity, stability, simple downstream purification, short period of fermentation and low cost. Earlier, we showed that Bacillus sp. IMV B-7883 strain synthesizes an extracellular proteases, which exhibit elastolytic and fibrinogenolytic activity. The aim of the work was to isolate and purify these enzymes from the culture liquid of the Bacillus sp. IMV B-7883 strain, as well as to study their properties. Isolation and purification of proteases was carried out by precipitation of the culture liquid with ammonium sulfate, gel permeation and ion exchange chromatography and rechromatography on Sepharose 6B. As a result, proteases with elastolytic and fibrinogenolytic activity with a molecular weight of 23 and 20 kDa respectively were isolated with elastase activity increased by 63.6 and fibrinogenolytic activity by 44.1 times. The enzyme with elastase activity had a pH-optimum of 7.0 and hydrolyzed only elastin, while the enzyme with fibrinogenolytic activity was an alkaline protease with a pH-optimum of 8.0 and in addition to fibrinogen, showed specificity for fibrin and, in trace amounts, for collagen.http://ukrbiochemjournal.org/wp-content/uploads/2023/11/Gudzenko_95_5.pdfbacillus sp. imv b-7883elastasefibrinogenaseph optimumsubstrate specificity |
| spellingShingle | O. V. Gudzenko L. D. Varbanets Isolation and characterization of Bacillus sp. IMV B-7883 proteases The Ukrainian Biochemical Journal bacillus sp. imv b-7883 elastase fibrinogenase ph optimum substrate specificity |
| title | Isolation and characterization of Bacillus sp. IMV B-7883 proteases |
| title_full | Isolation and characterization of Bacillus sp. IMV B-7883 proteases |
| title_fullStr | Isolation and characterization of Bacillus sp. IMV B-7883 proteases |
| title_full_unstemmed | Isolation and characterization of Bacillus sp. IMV B-7883 proteases |
| title_short | Isolation and characterization of Bacillus sp. IMV B-7883 proteases |
| title_sort | isolation and characterization of bacillus sp imv b 7883 proteases |
| topic | bacillus sp. imv b-7883 elastase fibrinogenase ph optimum substrate specificity |
| url | http://ukrbiochemjournal.org/wp-content/uploads/2023/11/Gudzenko_95_5.pdf |
| work_keys_str_mv | AT ovgudzenko isolationandcharacterizationofbacillusspimvb7883proteases AT ldvarbanets isolationandcharacterizationofbacillusspimvb7883proteases |