Purification and study of anti-cancer effects of
Anti-cancer
Ion exchange chromatography
Purification
Serralysin
Serratia marcescens
Journal
Iranian journal of microbiology
ISSN: 2008-3289
Titre abrégé: Iran J Microbiol
Pays: Iran
ID NLM: 101518404
Informations de publication
Date de publication:
Aug 2019
Aug 2019
Historique:
entrez:
14
11
2019
pubmed:
14
11
2019
medline:
14
11
2019
Statut:
ppublish
Résumé
Serralysin is an extracellular metalloprotease from The presence of the serralysin gene was confirmed using PCR. The supernatant of bacterial culture was collected and precipitated using ammonium sulfate. The precipitated protein was dialyzed and subjected to ion exchange chromatography for further purification. Casein assay and skim milk assay was used to confirm the enzymatic activity. SDS-PAGE was used to visualize the presence of serralysin. Metalloprotease inhibition activity was performed using 50 mM EDTA. Cytotoxic activity of serralysin was assessed on MTT assay. The PCR product corresponding to serralysin was estimated to be approximately 1500 bp. A transparent zone around the bacterial colonies on skim milk agar and casein digestion confirmed the proteolytic activity of serralysin. A 52 kDa band in SDS-PAGE corresponding to serralysin was observed before and after purification processes. MTT assay showed IC Our results showed that native serralysin has anticancer potential and may be a candidate for further pharmaceutical research and development. Further
Sections du résumé
BACKGROUND AND OBJECTIVES
OBJECTIVE
Serralysin is an extracellular metalloprotease from
MATERIALS AND METHODS
METHODS
The presence of the serralysin gene was confirmed using PCR. The supernatant of bacterial culture was collected and precipitated using ammonium sulfate. The precipitated protein was dialyzed and subjected to ion exchange chromatography for further purification. Casein assay and skim milk assay was used to confirm the enzymatic activity. SDS-PAGE was used to visualize the presence of serralysin. Metalloprotease inhibition activity was performed using 50 mM EDTA. Cytotoxic activity of serralysin was assessed on MTT assay.
RESULTS
RESULTS
The PCR product corresponding to serralysin was estimated to be approximately 1500 bp. A transparent zone around the bacterial colonies on skim milk agar and casein digestion confirmed the proteolytic activity of serralysin. A 52 kDa band in SDS-PAGE corresponding to serralysin was observed before and after purification processes. MTT assay showed IC
CONCLUSION
CONCLUSIONS
Our results showed that native serralysin has anticancer potential and may be a candidate for further pharmaceutical research and development. Further
Types de publication
Journal Article
Langues
eng
Pagination
320-327Informations de copyright
Copyright© 2019 Iranian Neuroscience Society.
Références
Methods Mol Biol. 2012;797:177-204
pubmed: 21948477
Singapore Med J. 1989 Feb;30(1):48-54
pubmed: 2688125
Sci Rep. 2016 Sep 08;6:32926
pubmed: 27605431
Sensors (Basel). 2012;12(3):2519-38
pubmed: 22736963
BMC Microbiol. 2015 Oct 09;15:207
pubmed: 26453184
J Vis Exp. 2008 Sep 17;(19):
pubmed: 19066538
Infect Agent Cancer. 2018 Mar 15;13:9
pubmed: 29568324
Int J Surg. 2013;11(3):209-17
pubmed: 23380245
Biochem Pharmacol. 2006 Apr 28;71(9):1289-98
pubmed: 16530733
Gastroenterology. 2010 Jun;138(6):2101-2114.e5
pubmed: 20420949
Arch Microbiol. 1980 Jan;124(1):55-61
pubmed: 6990888
Asian J Pharm Sci. 2017 May;12(3):209-215
pubmed: 32104332
Microb Pathog. 2013 Oct;63:44-53
pubmed: 23811076
CA Cancer J Clin. 2018 Nov;68(6):394-424
pubmed: 30207593
Curr Protoc Mol Biol. 2006 Aug;Chapter 10:Unit 10.2A
pubmed: 18265373
Curr Biol. 2012 Sep 11;22(17):R733-40
pubmed: 22975004
Food Chem Toxicol. 2016 Oct;96:79-89
pubmed: 27470613
Front Oncol. 2017 Sep 08;7:195
pubmed: 28944214
J Bacteriol. 1988 Sep;170(9):4141-6
pubmed: 2842305
J Int Med Res. 1990 Sep-Oct;18(5):379-88
pubmed: 2257960
Expert Opin Biol Ther. 2017 Aug;17(8):945-959
pubmed: 28604109