Functional evaluation of a recombinant fungal immunomodulatory protein from L. rhinocerus produced in P. pastoris and E. coli host expression systems

Characterization, Recombinant Production, and Bioactivity of a Novel Immunomodulatory Protein from Hypsizygus marmoreus

“…At present, genome mining has become a new tool for searching for potential FIPs. Ten FIPs, such as FIP-lrh [ 22 ], FIP-mco [ 35 ], and FIP-sch2 [ 36 ], have been directly cloned through genome mining. The low yield and high cost limit the extraction and application of natural FIPs.…”

“…The E. coli system is considered the host of choice for recombinant protein production due to fast growth speed, easy operation, and low cost, whose associated parameters (such as plasmids and incubation conditions) could be specifically optimized to improve the production capacity of the target protein [ 37 ]. Previously, several recombinant fungal immunomodulatory proteins, such as FIP-Lrh from L. rhinoceros [ 22 ], FIP-gat from Ganoderma atrum [ 38 ], and FIP-sch2 from Stachybotrys chlorohalonata [ 36 ] were successfully produced in E. coli and later showed efficient bioactivities. In Kong’s study [ 21 ], the production of bioactive FIP-fve in E. coli was six times higher than extracted from nature and has competent effects in promoting IFN-γ production in vivo.…”

“…It has been reported that FIPs derived from different fungi affect various cytokine profiles, where the results are also related to the dosage, molecular modification, or structure applied. For instance, rFIP-Lrh at lower concentrations (4–8 μg/mL) induced significantly higher production of Th1 (IFN-γ, TNF-α) and Th2 (IL-6, IL-4, IL-5, IL-13) cytokines in mice splenocytes, whereas 16 μg/mL rFIP-Lrh induced significantly higher pro-inflammatory cytokines (TNF-α, IL-6, IL-10) [ 22 ]. The findings on rFIP-hma favor that it triggered an acute immune response and activation of the central immunoregulatory networking, which were similar to the observed effects of several reported FIPs such as FIP-gsi (isolated from G. sinensis ) [ 13 ] and ACA (isolated from A. camphorate) [ 26 ].…”

“…FIP-fve was successfully expressed in E. coli , whose yield reached 29.1 mg/L with 97.1% purity; rFIP-fve regulated IL-2 and IFN-γ in mice, showing potential for industrial applications [ 21 ]. A recombinant fungal immunomodulatory protein from Lignosus rhinoceros was produced in P. pastoris and E. coli host expression systems, which (at 4–8 μg/mL doses) induced expression of Th1 (IFN-γ, TNF-α) and Th2 (IL-6, IL-4, IL-5, IL-13) cytokines in mice splenocytes [ 22 ]. A recombinant fungal immunomodulatory protein, GMI, was cloned from G. microsporum and purified, which inhibited migration and invasion of lung cancer cells, the mechanism was related to restraining activation of the EGFR and Akt pathway [ 23 ].…”