Abstract
Glucagon-like peptide-1 (GLP-1) is an incretin hormone that decreases postprandial glycemic excursions by enhancing insulin secretion but with short half-life due to rapid inactivation by enzymatic N-terminal truncation. Therefore, efforts are being made to improve the stability of GLP-1 via modifying its structure or inhibiting dipeptidylpeptidase IV (DPP IV), which is responsible for its degradation. GLP-M, consisting of 10 tandem repeated rolGLP-1 (GLP-1 analog), has been expressed in Pichia pastoris by our laboratory. Although it had a long effect of maintaining glucose homeostasis, redundant amino acids and purification tag limited its application. Here, optimized rolGLP-1(GLPO) with no redundant amino acids and purification tag was constructed by molecular cloning and site-directed mutagenesis, which was expressed efficiently in Escherichia coli BL21(DE3) with the production of 81.5 mg/L, and confirmed by the results of SDS-PAGE electrophoresis and Western Blotting. Then GLP-O was purified via ion exchange chromatography and gel filtration chromatography. The purity of GLP-O was close to 100%. GLP-O could be cut into single rolGLP-1 by trypsin in vitro, and rolGLP-1 had anti-trypsin activity. After oral administration of GLP-O for 4 weeks, the level of blood glucose in type 2 diabetic mice was lowered effectively, and the oral glucose tolerance of mice was improved significantly. These results settled the foundation for further clinical application of GLP-O.
Keywords: Diabetes, expression, GLP-1, Oral glucose tolerance, Purification, Western Blotting.
Current Pharmaceutical Biotechnology
Title:Expression and Purification of Optimized rolGLP-1, A Novel GLP-1 Analog, in Escherichia Coli BL21(DE3) and its Good Glucoregulatory Effect on Type 2 Diabetic Mice
Volume: 14 Issue: 11
Author(s): Baicheng Ma, Peipei Tu, Xingyu Zhao, Yaofang Zhang, Yu Wang, Chao Ma, Yanli Ji, Xiaodan Li, Syed A. Abbas and Minggang Li
Affiliation:
Keywords: Diabetes, expression, GLP-1, Oral glucose tolerance, Purification, Western Blotting.
Abstract: Glucagon-like peptide-1 (GLP-1) is an incretin hormone that decreases postprandial glycemic excursions by enhancing insulin secretion but with short half-life due to rapid inactivation by enzymatic N-terminal truncation. Therefore, efforts are being made to improve the stability of GLP-1 via modifying its structure or inhibiting dipeptidylpeptidase IV (DPP IV), which is responsible for its degradation. GLP-M, consisting of 10 tandem repeated rolGLP-1 (GLP-1 analog), has been expressed in Pichia pastoris by our laboratory. Although it had a long effect of maintaining glucose homeostasis, redundant amino acids and purification tag limited its application. Here, optimized rolGLP-1(GLPO) with no redundant amino acids and purification tag was constructed by molecular cloning and site-directed mutagenesis, which was expressed efficiently in Escherichia coli BL21(DE3) with the production of 81.5 mg/L, and confirmed by the results of SDS-PAGE electrophoresis and Western Blotting. Then GLP-O was purified via ion exchange chromatography and gel filtration chromatography. The purity of GLP-O was close to 100%. GLP-O could be cut into single rolGLP-1 by trypsin in vitro, and rolGLP-1 had anti-trypsin activity. After oral administration of GLP-O for 4 weeks, the level of blood glucose in type 2 diabetic mice was lowered effectively, and the oral glucose tolerance of mice was improved significantly. These results settled the foundation for further clinical application of GLP-O.
Export Options
About this article
Cite this article as:
Ma Baicheng, Tu Peipei, Zhao Xingyu, Zhang Yaofang, Wang Yu, Ma Chao, Ji Yanli, Li Xiaodan, Abbas A. Syed and Li Minggang, Expression and Purification of Optimized rolGLP-1, A Novel GLP-1 Analog, in Escherichia Coli BL21(DE3) and its Good Glucoregulatory Effect on Type 2 Diabetic Mice, Current Pharmaceutical Biotechnology 2013; 14 (11) . https://dx.doi.org/10.2174/1389201014666131226155553
DOI https://dx.doi.org/10.2174/1389201014666131226155553 |
Print ISSN 1389-2010 |
Publisher Name Bentham Science Publisher |
Online ISSN 1873-4316 |
Call for Papers in Thematic Issues
Artificial Intelligence in Bioinformatics
Bioinformatics is an interdisciplinary field that analyzes and explores biological data. This field combines biology and information system. Artificial Intelligence (AI) has attracted great attention as it tries to replicate human intelligence. It has become common technology for analyzing and solving complex data and problems and encompasses sub-fields of machine ...read more
- 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
-
Diabetic CVD – Focus on Vitamin D
Cardiovascular & Hematological Agents in Medicinal Chemistry Antidiabetic Oils
Current Diabetes Reviews Drugs in Newborn Resuscitation: The More We Learn the Least We Use
Current Medicinal Chemistry Autoimmune Hepatitis
Current Pediatric Reviews Association of Sleep Apnea Syndrome and Diabetes Mellitus
Current Respiratory Medicine Reviews Disorder in Milk Proteins: α -Lactalbumin. Part A. Structural Properties and Conformational Behavior
Current Protein & Peptide Science Consequences of Type 1 and 2 Diabetes Mellitus on the Cardiovascular Regulation During Exercise: A Brief Review
Current Diabetes Reviews Role of Endogenous GLP-1 and its Agonists in Osteopenia and Osteoporosis: But We Little Know Until Tried
Current Diabetes Reviews Intramolecular Cyclisation of β-Aryl-β-Amino Acids in the Design of Novel Heterocyclic Systems with Therapeutic Interest: An Unfailing Source of Diversity
Current Medicinal Chemistry The Association of Transcription Factor 7 like 2 Gene Polymorphism with Diabetic Nephropathy in Patients with Type 2 Diabetes Mellitus
Current Diabetes Reviews The Role of Amino Acids in the Modulation of Cardiac Metabolism During Ischemia and Heart Failure
Current Pharmaceutical Design Human Clinical Studies of Tea Polyphenols in Allergy or Life Style-related Diseases
Current Pharmaceutical Design Cardiovascular Disease Risk of Type 2 Diabetes Mellitus and Metabolic Syndrome: Focus on Aggressive Management of Dyslipidemia
Current Diabetes Reviews Chemoresistance in High-Grade Gliomas: Relevance of Adenosine Signalling in Stem-Like Cells of Glioblastoma Multiforme
Current Drug Targets A Hybrid Binary Cuckoo Search and Genetic Algorithm for Feature Selection in Type-2 Diabetes
Current Bioinformatics Oxidative Stress Correlates (OSC) in Diabetes Mellitus Patients
Current Diabetes Reviews Amphetamine-Type Medicines: A Review of Pharmacokinetics, Pharmacodynamics, and Toxicological Aspects.
Current Clinical Pharmacology Cholesterol Lowering Therapy: Treat to Target or Reduce the Global Risk? The Unresolved Problem of Residual Risk
Current Pharmaceutical Design Lysophosphatidic Acid and Renal Fibrosis
Recent Patents on Endocrine, Metabolic & Immune Drug Discovery Therapeutic Nanosystems for Oral Administration of Insulin
Current Pharmaceutical Biotechnology