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Biochemical Characterization of An Arginine-Specific Alkaline Trypsin from Bacillus licheniformis
- Source :
- International Journal of Molecular Sciences, International Journal of Molecular Sciences; Volume 16; Issue 12; Pages: 30061-30074, International Journal of Molecular Sciences, Vol 16, Iss 12, Pp 30061-30074 (2015), Volume 16, Issue 12, Pages 30061-30074
- Publication Year :
- 2015
- Publisher :
- MDPI, 2015.
-
Abstract
- In the present study, we isolated a trypsin-producing strain DMN6 from the leather waste and identified it as Bacillus licheniformis through a two-step screening strategy. The trypsin activity was increased up to 140 from 20 U/mL through culture optimization. The enzyme was purified to electrophoretic homogeneity with a molecular mass of 44 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the specific activity of purified enzyme is 350 U/mg with Nα-Benzoyl-L-arginine ethylester as the substrate. The optimum temperature and pH for the trypsin are 65 °C and pH 9.0, respectively. Also, the enzyme can be significantly activated by Ba(2+). This enzyme is relatively stable in alkaline environment and displays excellent activity at low temperatures. It could retain over 95% of enzyme activity after 180 min of incubation at 45 °C. The distinguished activity under low temperature and prominent stability enhance its catalytic potential. In the current work, the open reading frame was obtained with a length of 1371 nucleotides that encoded a protein of 456 amino acids. These data would warrant the B. licheniformis trypsin as a promising candidate for catalytic application in collagen preparation and leather bating through further protein engineering.
- Subjects :
- Bacillus
Alkalies
Bacillus licheniformis
Substrate Specificity
lcsh:Chemistry
RNA, Ribosomal, 16S
Enzyme Stability
Trypsin
Cloning, Molecular
lcsh:QH301-705.5
Spectroscopy
Phylogeny
Gel electrophoresis
chemistry.chemical_classification
biology
Temperature
General Medicine
Hydrogen-Ion Concentration
Computer Science Applications
Biochemistry
Metals
trypsin
enzymatic properties
cloning
biocatalysis
Electrophoresis, Polyacrylamide Gel
Trypsin Inhibitors
medicine.drug
Molecular Sequence Data
Arginine
Catalysis
Article
Inorganic Chemistry
medicine
Amino Acid Sequence
Physical and Theoretical Chemistry
Molecular Biology
Ions
Molecular mass
Base Sequence
Organic Chemistry
Substrate (chemistry)
biology.organism_classification
Enzyme assay
Kinetics
Enzyme
chemistry
lcsh:Biology (General)
lcsh:QD1-999
Genes, Bacterial
Fermentation
biology.protein
Specific activity
Sequence Alignment
Subjects
Details
- Language :
- English
- ISSN :
- 14220067
- Volume :
- 16
- Issue :
- 12
- Database :
- OpenAIRE
- Journal :
- International Journal of Molecular Sciences
- Accession number :
- edsair.doi.dedup.....239d2d4b74bad7d44201f4a49161ecd3