Identification a novel Ganoderma FIP gene from Ganoderma capense and its functional expression in Pichia pastoris

In this study, a novel fungal immunomodulatory protein gene, named as FIP-gca, was cloned fromG. capense by homologous cloning. Sequencing analysis indicated that FIP-gca was composed of 336  bp, which encoded a polypeptide of 110 amino acids. Protein sequence blasting and phylogenetic analysis showed that FIP-gca shared homology with otherGanoderma FIPs.FIP-gca was effectively expressed inPichia pastoris GS115 at an expression level of 166.8  mg/L and purified using HisTrap™ fast-flow prepack columns. The immunomodulation capacity of rFIP-gca was demonstrated by that rFIP-gca could obviously stimulate cell proliferation and increase IL-2 secretion of murine spleen lymphocytes. Besides, antitumor activity of rFIP-gca towards human sto mach cancer AGS cell line was evaluated in vitro. Cell wound scratch assay proved that rFIP-gca could inhibit migration of AGS cells. And flow cytometry assay revealed that rFIP-gca could significantly induce apoptosis of AGS cells. rFIP-gca was able to induce 18.12% and 22.29% cell apoptosis at 0.3  μM and 0.6 μM, respectively. Conclusively, the novelFIP-gca gene fromG. capense has been functionally expressed inPichia and rFIP-gca exhibited ideal immunomodulation and anti-tumour activities, which implies its potential application and study in future.
Source: World Journal of Microbiology and Biotechnology - Category: Microbiology Source Type: research