Abstract
Proteases hydrolyze polypeptides to release peptides and/or amino acids. This subclass of enzymes is among those with the most sales worldwide, particularly those produced by microorganisms. Proteases may be applied in the several industries, including the food industry, leather, detergents, and bioremediation. Myceliophthora thermophila protease was produced by a submerged bioprocess and then purified 185-fold by anion exchange and hydrophobic chromatography with a 37% yield. The molecular mass was estimated at 36.2 kDa, and mass spectrometry identified two sequences: GVVANMSLGGSYSASINNAAAALVR and STGNAAITGVPSGTTNR. The isolated protein was characterized biochemically, showed an optimum pH of 6.5 and optimum temperature of 45 °C, and stability at wide range of pH and temperatures and in the presence of reducing agents and some surfactants. Kinetic assays for this enzyme showed a greater catalytic efficiency when the substrate had alanine at position P'2. The protease presented characteristics that may be of interest to many industrial areas.
Keywords: FRET substrates, neutral protease, mass spectrometry, Myceliophthora thermophila, purification, stability.
Protein & Peptide Letters
Title:Determination of Specificity and Biochemical Characteristics of Neutral Protease Isolated from Myceliophthora thermophila
Volume: 22 Issue: 11
Author(s): Youssef A.A. Hamin Neto, Lilian C.G. de Oliveira, Arthur H.C. de Oliveira, Jose C. Rosa, Maria A. Juliano, Luiz Juliano, Andre Rodrigues and Hamilton Cabral
Affiliation:
Keywords: FRET substrates, neutral protease, mass spectrometry, Myceliophthora thermophila, purification, stability.
Abstract: Proteases hydrolyze polypeptides to release peptides and/or amino acids. This subclass of enzymes is among those with the most sales worldwide, particularly those produced by microorganisms. Proteases may be applied in the several industries, including the food industry, leather, detergents, and bioremediation. Myceliophthora thermophila protease was produced by a submerged bioprocess and then purified 185-fold by anion exchange and hydrophobic chromatography with a 37% yield. The molecular mass was estimated at 36.2 kDa, and mass spectrometry identified two sequences: GVVANMSLGGSYSASINNAAAALVR and STGNAAITGVPSGTTNR. The isolated protein was characterized biochemically, showed an optimum pH of 6.5 and optimum temperature of 45 °C, and stability at wide range of pH and temperatures and in the presence of reducing agents and some surfactants. Kinetic assays for this enzyme showed a greater catalytic efficiency when the substrate had alanine at position P'2. The protease presented characteristics that may be of interest to many industrial areas.
Export Options
About this article
Cite this article as:
Hamin Neto A.A. Youssef, de Oliveira C.G. Lilian, de Oliveira H.C. Arthur, Rosa C. Jose, Juliano A. Maria, Juliano Luiz, Rodrigues Andre and Cabral Hamilton, Determination of Specificity and Biochemical Characteristics of Neutral Protease Isolated from Myceliophthora thermophila, Protein & Peptide Letters 2015; 22 (11) . https://dx.doi.org/10.2174/0929866522666150817093719
DOI https://dx.doi.org/10.2174/0929866522666150817093719 |
Print ISSN 0929-8665 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5305 |
- Author Guidelines
- Graphical Abstracts
- Fabricating and Stating False Information
- Research Misconduct
- Post Publication Discussions and Corrections
- Publishing Ethics and Rectitude
- Increase Visibility of Your Article
- Archiving Policies
- Peer Review Workflow
- Order Your Article Before Print
- Promote Your Article
- Manuscript Transfer Facility
- Editorial Policies
- Allegations from Whistleblowers
Related Articles
-
Adult Neural Stem Cell Therapy: Expansion In Vitro, Tracking In Vivo and Clinical Transplantation
Current Drug Targets Synthesis of Arylpiperazine Derivatives as Protease Activated Receptor 1 Antagonists and Their Evaluation as Antiproliferative Agents
Anti-Cancer Agents in Medicinal Chemistry Nucleic Acid Carrier Systems Based on Polyethylenimine Conjugates for the Treatment of Metastatic Tumors
Current Medicinal Chemistry Drug Metabolism and Transport Under Hypoxia
Current Drug Metabolism Integrins in Bone Metastasis Formation and Potential Therapeutic Implications
Current Cancer Drug Targets Understanding Autophagy in Cell Death Control
Current Pharmaceutical Design Antiangiogenic Resistance: Novel Angiogenesis Axes Uncovered by Antiangiogenic Therapies Research
Current Drug Targets Marine Derived Anticancer Drugs Targeting Microtubule
Recent Patents on Anti-Cancer Drug Discovery Molecular Chaperone ORP150 in ER Stress–related Diseases
Current Pharmaceutical Design The Emerging Role of EMT-related lncRNAs in Therapy Resistance and their Applications as Biomarkers
Current Medicinal Chemistry Combination of Phytochemicals as Adjuvants for Cancer Therapy
Recent Patents on Anti-Cancer Drug Discovery Will Arsenic Trioxide Benefit Treatment of Solid Tumor by Nano- Encapsulation?
Mini-Reviews in Medicinal Chemistry Recent Progress on Antifungal Drug Development
Current Pharmaceutical Biotechnology Cell Metabolism Under Microenvironmental Low Oxygen Tension Levels in Stemness, Proliferation and Pluripotency
Current Molecular Medicine Approaches to Improve Efficiency of Dendritic Cell-based Therapy of High Grade Gliomas
Current Pharmaceutical Design Astrocytes: Implications for Neuroinflammatory Pathogenesis of Alzheimers Disease
Current Alzheimer Research Modulation of MicroRNAs by <i>Euphorbia Microsciadia</i> Boiss in MDA-MB-231 Cell Line: New Possibilities in Breast Cancer Therapy
Recent Patents on Anti-Cancer Drug Discovery The Molecular Mechanisms of TRAIL Resistance in Cancer Cells: Help in Designing New Drugs
Current Pharmaceutical Design Chronic Inflammation and Colorectal Cancer: The Role of Vascular Endothelial Growth Factor
Current Pharmaceutical Design Dendritoma Vaccine for Cancer: A Hopeful Approach
Current Cancer Therapy Reviews