Back to Search
Start Over
Impact of signal peptide and transmembrane segments on expression and biochemical properties of a lipase from Bacillus sphaericus 205y
- Source :
- Journal of Biotechnology. 264:51-62
- Publication Year :
- 2017
- Publisher :
- Elsevier BV, 2017.
-
Abstract
- A total of 97 amino acids, considered as the signal peptide and transmembrane segments were removed from 205y lipase gene using polymerase chain reaction technique that abolished the low activity of this enzyme. The mature enzyme was expressed in Escherichia coli using pBAD expression vector, which gave up to a 13-fold increase in lipase activity. The mature 205y lipase (without signal peptide and transmembrane; -SP/TM) was purified to homogeneity using the isoelectric focusing technique with 53% recovery. Removing of the signal peptide and transmembrane segments had resulted in the shift of optimal pH, an increase in optimal temperature and tolerance towards more water-miscible organic solvents as compared to the characteristics of open reading frame (ORF) of 205y lipase. Also, in the presence of 1mM inhibitors, less decrease in the activity of mature 205y lipase was observed compared to the ORF of the enzyme. Protein structure modeling showed that 205y lipase consisted of an α/β hydrolase fold without lid domain. However, the transmembrane segment could effect on the enzyme activity by covering the active site or aggregation the protein.
- Subjects :
- 0106 biological sciences
0301 basic medicine
Signal peptide
Bacillus
Bioengineering
Protein Sorting Signals
01 natural sciences
Applied Microbiology and Biotechnology
Substrate Specificity
03 medical and health sciences
Bacterial Proteins
010608 biotechnology
Hydrolase
Escherichia coli
Cloning, Molecular
Lipase
Triglycerides
chemistry.chemical_classification
biology
Membrane Proteins
General Medicine
Molecular biology
Recombinant Proteins
Transmembrane protein
Enzyme assay
Molecular Docking Simulation
Transmembrane domain
Open reading frame
030104 developmental biology
Enzyme
chemistry
Biochemistry
biology.protein
Oils
Biotechnology
Subjects
Details
- ISSN :
- 01681656
- Volume :
- 264
- Database :
- OpenAIRE
- Journal :
- Journal of Biotechnology
- Accession number :
- edsair.doi.dedup.....777e097ff939bf208135f6a32cea0f5a