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Updated: Mar 3, 2026
Expression of Recombinant Proteins in the Methylotrophic Yeast Pichia pastoris
Published on: February 25, 2010
Heterologous expression of abaecin peptide from Apis mellifera in Pichia pastoris
Denis Prudencio Luiz1, Juliana Franco Almeida2, Luiz Ricardo Goulart2
1Genetics Laboratory, Institute of Genetics and Biochemistry, Federal University of Uberlândia, 1720 Pará, Uberlândia, MG, 38400902, Brazil. prudenis@yahoo.com.br.
Background:
Antimicrobial peptides (AMPs) are the first line of host immune defense against pathogens. Among AMPs from the honeybee Apis mellifera, abaecin is a major broad-spectrum antibacterial proline-enriched cationic peptide.
Results:
For heterologous expression of abaecin in Pichia pastoris, we designed an ORF with HisTag, and the codon usage was optimized. The gene was chemically synthetized and cloned in the pUC57 vector. The new ORF was sub-cloned in the pPIC9 expression vector and transformed into P. pastoris. After selection of positive clones, the expression was induced by methanol. The supernatant was analyzed at different times to determine the optimal time for the recombinant peptide expression. As a proof-of-concept, Escherichia coli was co-incubated with the recombinant peptide to verify its antimicrobial potential.
Discussion:
Briefly, the recombinant Abaecin (rAbaecin) has efficiently decreased E. coli growth (P < 0.05) through an in vitro assay, and may be considered as a novel therapeutic agent that may complement other conventional antibiotic therapies.
Insights
Honeybee abaecin, a broad-spectrum antimicrobial peptide, was successfully expressed in Pichia pastoris. The recombinant abaecin (rAbaecin) demonstrated significant antibacterial activity against E. coli in vitro.
Area of Science:
- Biochemistry
- Molecular Biology
- Microbiology
Background:
- Antimicrobial peptides (AMPs) are crucial for innate immunity against pathogens.
- Abaecin, a proline-rich cationic peptide from the honeybee Apis mellifera, exhibits broad-spectrum antibacterial properties.
Purpose of the Study:
- To achieve heterologous expression of abaecin in Pichia pastoris.
- To evaluate the antimicrobial potential of recombinant abaecin (rAbaecin).
Main Methods:
- Designed and synthesized an optimized codon usage Open Reading Frame (ORF) for abaecin with a HisTag.
- Cloned the ORF into pUC57 and subsequently into the pPIC9 expression vector for transformation into P. pastoris.
- Induced recombinant protein expression using methanol and analyzed supernatant for rAbaecin production.
- Verified antimicrobial activity through in vitro co-incubation assays with Escherichia coli.
Main Results:
- Successfully expressed and purified recombinant abaecin (rAbaecin) in Pichia pastoris.
- Demonstrated that rAbaecin significantly reduced Escherichia coli growth in vitro (P < 0.05).
Conclusions:
- Recombinant abaecin shows potent antimicrobial activity against E. coli.
- rAbaecin holds promise as a novel therapeutic agent to supplement existing antibiotic treatments.
