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Expression and characterization of a novel glycoside hydrolase family 46 chitosanase identified from marine mud metagenome
- Source :
- International Journal of Biological Macromolecules. 159:904-910
- Publication Year :
- 2020
- Publisher :
- Elsevier BV, 2020.
-
Abstract
- A novel chitosanase gene, csn4, was identified through function-based screening of a marine mud metagenomic library. The encoded protein, named CSN4, which belonged to glycoside hydrolase family 46, showed its maximum identity (79%) with Methylobacter tundripaludum peptidoglycan-binding protein. CSN4 was expressed in Escherichia coli and purified. It displayed maximal activity at 30 °C and pH 7. A weakly-alkaline solution strongly inhibited the activity. The enzymatic activity was enhanced by addition of Mn2+ or Co2+. CSN4 exhibited strict substrate specificity for chitosan, and the activity was enhanced by increasing the degree of deacetylation. Thin-layer chromatography and electrospray ionization-mass spectrometry showed that CSN4 displayed an endo-type cleavage pattern, hydrolyzing chitosan mainly into (GlcN)2, (GlcN)3 and (GlcN)4. The novel characteristics of the chitosanase CSN4 make it a potential candidate to produce chitooligosaccharides from chitosan in industry.
- Subjects :
- Aquatic Organisms
Spectrometry, Mass, Electrospray Ionization
Chemical Phenomena
Glycoside Hydrolases
Gene Expression
02 engineering and technology
medicine.disease_cause
Biochemistry
Substrate Specificity
Chitosan
03 medical and health sciences
chemistry.chemical_compound
Structural Biology
medicine
Glycoside hydrolase
Amino Acid Sequence
Chitosanase
Cloning, Molecular
Molecular Biology
Gene
Escherichia coli
Phylogeny
030304 developmental biology
chemistry.chemical_classification
0303 health sciences
Hydrolysis
Methylobacter tundripaludum
Sequence Analysis, DNA
General Medicine
021001 nanoscience & nanotechnology
Enzyme Activation
carbohydrates (lipids)
Enzyme
chemistry
Acetylation
Multigene Family
Metagenome
Chromatography, Thin Layer
Metagenomics
0210 nano-technology
Subjects
Details
- ISSN :
- 01418130
- Volume :
- 159
- Database :
- OpenAIRE
- Journal :
- International Journal of Biological Macromolecules
- Accession number :
- edsair.doi.dedup.....1c8016ffc22427f7f88506ed6d4e0562