Design and cloning of tanatin peptide in pichia.pastoris and evaluation of its antifungal activity

Abbas Tanhaeian,1,*

Abstract


Introduction

Nowadays, control of fungal infections is one of the most important goals in the field of health. therefore, developing new and effective methods for inhibiting fungal pathogens is one of the main goals. tanatin is a potent antimicrobial peptide in inhibiting some of the fungal species, the natural source of which is a hunter insect that is used in the united states in its biologic struggle. in the present study, p.pastoris yeast was used as a production platform for the production of tannin peptide due to recombinant synthesis of yeast with secretory signal, the subsequent synthesis of tanatin by induction of expression with ethanol, secretion of it in the culture medium, and from a culture medium containing tanatin antifungal evaluation was used and the results showed that effective control of candida .albicans and candida. parapsilosis and mushroom mucus, isolated from the patient in qa'im hospital, were isolated.

Methods

The resulting nucleotide sequence was codonized optimization and then ppiczα a vector shear sites added to it and, after ensuring that it was within the framework of the reading, proceeded to synthesize the nucleotide sequencing, and after digestion and extraction of the gel and cutting vector of the expression vector with the same enzymes a ligation reaction was conducted for recombinant linearity for electroporation and induction of methanol expression every 6 hours.

Results

The recombinant vector was approved by a specific primer and the media was used to evaluate the accuracy of the expression and secretion and disk diffusion test to evaluate the antifungal function

Conclusion

The results of the evaluation of candida albicans and candida parapsylosis indicated that by increasing the recombinant tanatine, the diameter of the halo also, in the case of mucurium fungus in comparison with the non-tanatin medium as a control and a recombinant tanatin culture medium, the presence of tanatin inhibitor on these fungi is considered.

Keywords

Tanatin,recombinant,secretion,antifungal,pichia.pastoris