Identification and Heterologous Expression of the Biosynthetic Gene Cluster Encoding the Lasso Peptide Humidimycin, a
Marina Sánchez-Hidalgo1, Jesús Martín1, Olga Genilloud1
1Fundación MEDINA, Avenida del Conocimiento 34, 18016 Granada, Spain.
Abstract:
Humidimycin (MDN-0010) is a ribosomally synthesized and post-translationally modified peptide (RiPP) belonging to class I lasso peptides, and is structurally related to siamycins, which have been shown to have strong antimicrobial activities against Gram-positive bacteria and to possess anti-HIV activity. Humidimycin was isolated from the strain Streptomyces humidus CA-100629, and was shown to synergize the activity of the fungal cell wall inhibitor caspofungin. In this work, the biosynthetic gene cluster of humidimycin was identified by genome mining of S. humidus CA-100629, cloned by Gibson assembly, and heterologously expressed.
Insights
Humidimycin, a lasso peptide antibiotic, shows potential antimicrobial and anti-HIV activities. Researchers identified and cloned its biosynthetic gene cluster from Streptomyces humidus for further study.
Area of Science:
- Microbiology
- Molecular Biology
- Biochemistry
Background:
- Humidimycin (MDN-0010) is a class I lasso peptide with structural similarity to siamycins.
- Siamycins exhibit potent antimicrobial activity against Gram-positive bacteria and anti-HIV properties.
- Humidimycin synergizes with caspofungin, a fungal cell wall inhibitor.
Purpose of the Study:
- To identify the biosynthetic gene cluster of humidimycin.
- To clone the identified gene cluster for heterologous expression.
- To enable further investigation into humidimycin's biosynthesis and potential applications.
Main Methods:
- Genome mining of *Streptomyces humidus* CA-100629 to locate the humidimycin gene cluster.
- Cloning of the gene cluster using Gibson assembly.
- Heterologous expression of the cloned gene cluster.
Main Results:
- The biosynthetic gene cluster responsible for humidimycin production was successfully identified in *S. humidus*.
- The gene cluster was cloned and prepared for heterologous expression.
- This work lays the foundation for producing humidimycin in a heterologous host.
Conclusions:
- The identification and cloning of the humidimycin biosynthetic gene cluster represent a significant step towards understanding and producing this promising RiPP.
- Heterologous expression will facilitate detailed mechanistic studies and potential engineering for novel antimicrobial agents.
- This research opens avenues for developing new therapeutic strategies leveraging humidimycin's unique properties.
