Abstract
The expression in Escherichia coli strain Rosetta of the recombinant acidic subunit from the 11S amaranth seed storage protein (protein ACM3) was studied at flask and at bioreactor levels. This subunit was modified by inserting four Val-Tyr antihypertensive peptides in tandem into its third variable region and also with the tripeptide Ile-Pro-Pro in the Cterminal region. Flasks experiments allowed us to define the best conditions for the preparation and expression and accumulation of the protein ACM3, including the certainty of its presence within the cells especially as an insoluble fraction. The effects of cultivation temperature, aeration rate and agitation speed on the production of the protein ACM3 was tested in a 5-L batch bioreactor. Applying response surface methodology (RSM) we found that the aeration rate was the most significant factor affecting in a positively way the production yields and productivity of the recombinant protein. Temperature had effect only in conjunction to aeration. The highest recombinant acidic subunit concentration (747 mg L-1) and the highest productivity (186 mg L-1 h-1) were attained in 4 h of cultivation when the factors evaluated were controlled at its central values: 0.1 vvm, 300 rpm, and 30.5°C. Results from this study indicate that RSM is an effective technique to maximize the production of this recombinant protein.
Keywords: E. coli-expressed protein, protein ACM3, batch fermentation, environmental conditions, response surface methodology.
Recent Patents on Biotechnology
Title:Effect of Environmental Conditions on the Expression Levels of a Recombinant 11S Amaranth Globulin in Escherichia coli
Volume: 6 Issue: 1
Author(s): Hypatia Arano-Varela, Jorge Dominguez-Dominguez and Octavio Paredes-Lopez
Affiliation:
Keywords: E. coli-expressed protein, protein ACM3, batch fermentation, environmental conditions, response surface methodology.
Abstract: The expression in Escherichia coli strain Rosetta of the recombinant acidic subunit from the 11S amaranth seed storage protein (protein ACM3) was studied at flask and at bioreactor levels. This subunit was modified by inserting four Val-Tyr antihypertensive peptides in tandem into its third variable region and also with the tripeptide Ile-Pro-Pro in the Cterminal region. Flasks experiments allowed us to define the best conditions for the preparation and expression and accumulation of the protein ACM3, including the certainty of its presence within the cells especially as an insoluble fraction. The effects of cultivation temperature, aeration rate and agitation speed on the production of the protein ACM3 was tested in a 5-L batch bioreactor. Applying response surface methodology (RSM) we found that the aeration rate was the most significant factor affecting in a positively way the production yields and productivity of the recombinant protein. Temperature had effect only in conjunction to aeration. The highest recombinant acidic subunit concentration (747 mg L-1) and the highest productivity (186 mg L-1 h-1) were attained in 4 h of cultivation when the factors evaluated were controlled at its central values: 0.1 vvm, 300 rpm, and 30.5°C. Results from this study indicate that RSM is an effective technique to maximize the production of this recombinant protein.
Export Options
About this article
Cite this article as:
Arano-Varela Hypatia, Dominguez-Dominguez Jorge and Paredes-Lopez Octavio, Effect of Environmental Conditions on the Expression Levels of a Recombinant 11S Amaranth Globulin in Escherichia coli, Recent Patents on Biotechnology 2012; 6 (1) . https://dx.doi.org/10.2174/187220812799789181
DOI https://dx.doi.org/10.2174/187220812799789181 |
Print ISSN 1872-2083 |
Publisher Name Bentham Science Publisher |
Online ISSN 2212-4012 |
Call for Papers in Thematic Issues
Advancements in Plant Genetics and Genomics
This thematic issue will highlight cutting-edge innovations and recent patents across the breadth of plant genetics and genomics. Key topics will include advanced breeding techniques, omics-based approaches, gene editing, trait discovery, and translational applications for crop improvement. Renowned researchers from leading institutions worldwide will provide state-of-the-art insights and future perspectives ...read more
Related Journals
- 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
-
Cerebrovascular Diseases in HIV-Infected Patients
Current HIV Research Editorial [Hot Topic:Modifying Cardiovascular Risk Factors: Newer Insights and Preventive Measures (Executive Editor: Aurelio Leone)]
Current Pharmaceutical Design ACE Inhibition, p300, Serum Nitrite and Lipid Peroxidation in Newly Diagnosed Hypertensives
Vascular Disease Prevention (Discontinued) Life-Threatening Respiratory Syncytial Virus Disease in Children
Current Respiratory Medicine Reviews Bidirectional Nature of Cardiovascular and Kidney Disease
Immunology, Endocrine & Metabolic Agents in Medicinal Chemistry (Discontinued) How Recent Patents Have Changed our Clinical Approach in Cardio-Thoracic Surgery
Recent Patents on Regenerative Medicine MCM-41-SO3H as an Advanced Nanocatalyst for the Solvent-Free Synthesis of N-aryl-2-amino-1,6-naphthyridine Derivatives
Current Nanoscience Homocysteine and Hyperhomocysteinaemia
Current Medicinal Chemistry Introduction from the Guest Editor: The Many Facets of Heme Oxygenase
Current Neurovascular Research Adenosine Receptors: New Therapeutic Targets for Inflammation in Diabetic Nephropathy
Inflammation & Allergy - Drug Targets (Discontinued) Mixed Connective Tissue Disease (MCTD) – A Coming of Age
Current Rheumatology Reviews Organoselenium Compounds as Potential Neuroprotective Therapeutic Agents
Current Organic Chemistry Thresholds for Central Blood Pressures and Augmentation Indices - are They Needed and How Far are We in the Process of Their Definition?
Current Hypertension Reviews Prediabetic Dysglycemia: Call for Action
Current Diabetes Reviews The Two fACEs of the Tissue Renin-Angiotensin Systems: Implication in Cardiovascular Diseases
Current Pharmaceutical Design Preface [Hot Topic: Treatment of Hypertension, Its Pathophysiology and the Clinical Use of Antihypertensive Drugs (Executive Editor: Gregory Y.H. Lip)]
Current Pharmaceutical Design Cardiovascular Side Effects of New Antidepressants and Antipsychotics: New Drugs, old Concerns?
Current Pharmaceutical Design Aspirin Resistance in Cardiovascular Disease: Pathogenesis, Diagnosis and Clinical Impact
Current Pharmaceutical Design Potential Use of Vanadium Compounds in Therapeutics
Current Medicinal Chemistry Gender Differences in the Effects of Angiotensin Receptor Blockers on Cardiovascular Disease
Current Pharmaceutical Design