Related Experiment Video
Updated: Mar 2, 2026
Gastric Mucosa Quantitative Polymerase Chain Reaction Analysis for Detecting Helicobacter pylori and Antibiotic Resistance
Published on: March 7, 2025
Fusion expression of the PGLa-AM1 with native structure and evaluation of its anti-Helicobacter pylori activity
Xiaolin Zhang1, Anmin Jiang2, Guisheng Wang3
1The Department of Pharmacy, Food and Drug School, Anhui Science and Technology University, Fengyang, China. zhangxiaolin8@126.com.
Abstract:
Helicobacter pylori (H. pylori) shows increasingly enhanced resistance to various antibiotics, and its eradication has become a major problem in medicine. The antimicrobial peptide PGLa-AM1 is a short peptide with 22 amino acids and exhibits strong antibacterial activity. In this study, we investigated whether it has anti-H. pylori activity for the further development of anti-H. pylori drugs to replace existing antibiotics. However, the natural antimicrobial peptide PGLa-AM1 shows a low yield and is difficult to separate, limiting its application. A good strategy to solve this problem is to express the antimicrobial peptide PGLa-AM1 using gene engineering at a high level and low cost. For getting PGLa-AM1 with native structure, in this study, a specific protease cleavage site of tobacco etch virus (TEV) was designed before the PGLa-AM1 peptide. For convenience to purify and identify high-efficiency expression PGLa-AM1, the PGLa-AM1 gene was fused with the polyhedrin gene of Bombyx mori (B. mori), and a 6 × His tag was designed to insert before the amino terminus of the fusion protein. The fusion antibacterial peptide PGLa-AM1 (FAMP) gene codon was optimized, and the gene was synthesized and cloned into the Escherichia coli (E. coli) pET-30a (+) expression vector. The results showed that the FAMP was successfully expressed in E. coli. Its molecular weight was approximately 34 kDa, and its expression level was approximately 30 mg/L. After the FAMP was purified, it was further digested with TEV protease. The acquired recombinant antimicrobial peptide PGLa-AM1 exerted strong anti-H. pylori activity and therapeutic effect in vitro and in vivo.
Insights
Researchers engineered a fusion protein for high-level production of the antimicrobial peptide PGLa-AM1. This peptide demonstrated potent activity against antibiotic-resistant Helicobacter pylori, offering a promising alternative for H. pylori eradication therapies.
Area of Science:
- Biotechnology
- Microbiology
- Drug Discovery
Background:
- Helicobacter pylori (H. pylori) infections are increasingly resistant to antibiotics, complicating eradication efforts.
- The antimicrobial peptide PGLa-AM1 shows potent antibacterial activity but suffers from low natural yield and difficult purification.
- Gene engineering offers a viable strategy for high-level, cost-effective production of PGLa-AM1.
Purpose of the Study:
- To develop a method for high-level expression of the antimicrobial peptide PGLa-AM1.
- To evaluate the anti-H. pylori activity of the recombinant PGLa-AM1.
- To explore PGLa-AM1 as a potential therapeutic agent against antibiotic-resistant H. pylori.
Main Methods:
- Engineered a fusion gene encoding PGLa-AM1 with a tobacco etch virus (TEV) protease cleavage site and a 6xHis tag, fused to the Bombyx mori polyhedrin gene.
- Optimized the fusion antibacterial peptide (FAMP) gene codon and expressed it in Escherichia coli (E. coli).
- Purified the expressed FAMP, cleaved PGLa-AM1 using TEV protease, and assessed its activity in vitro and in vivo.
Main Results:
- Successfully expressed the FAMP in E. coli with a molecular weight of approximately 34 kDa and an expression level of about 30 mg/L.
- Purified FAMP and obtained recombinant PGLa-AM1 after TEV protease digestion.
- Demonstrated strong in vitro and in vivo anti-H. pylori activity of the recombinant PGLa-AM1.
Conclusions:
- High-level expression and purification of PGLa-AM1 were achieved using a gene engineering approach.
- Recombinant PGLa-AM1 exhibits significant antibacterial activity against H. pylori.
- This strategy provides a promising avenue for developing novel anti-H. pylori drugs to combat antibiotic resistance.
More Related Videos
Related Concept Videos
01:16Treating Helicobacter pylori in Peptic Ulcers: Antimicrobial Therapy
01:28Peptic Ulcer Disease III: Clinical Manifestations and Diagnostic Studies
Few clinical manifestations differentiate gastric ulcers from duodenal ulcers. Distinctions in the location, timing, and pain relief are crucial for healthcare providers in differentiating between gastric and duodenal ulcers during clinical assessments.