Heterologous expression of the immunomodulatory protein gene from Ganoderma sinense in the basidiomycete Coprinopsis cinerea

Abstract

Aims: FIP‐gsi, a fungal immunomodulatory protein found in Ganoderma sinense, has antitumour, anti‐allergy and immunomodulatory activities and is regulated by the fip‐gsi gene. In this study, we aimed to express the fip‐gsi gene from G. sinense in Coprinopsis cinerea to increase yield of FIPs‐gsi. Methods and Results: A fungal expression vector pBfip‐gsi containing the gpd promoter from Agaricus bisporus and the fip‐gsi gene from the G. sinense was constructed and transformed into C. cinerea. PCR and Southern blotting analysis verified the successful integration of the exogenous gene fip‐gsi into the genome of C. cinerea. RT‐PCR and Northern blotting analysis confirmed that the fip‐gsi gene was transcribed in C. cinerea. The yield of the FIP‐gsi protein reached 314 mg kg−1 fresh mycelia. The molecular weight of the FIP‐gsi was 13 kDa, and the FIP‐gsi was capable of hemagglutinating mouse red blood cells, but no such activity was observed towards human red blood cells in vitro. Conclusions: The fip‐gsi from G. sinense has been successfully translated in C. cinerea, and the yield of bioactive FIP‐gsi protein was high. Significance and Impact of the Study: This is the first report using the C. cinerea for the heterologous expression of FIP‐gsi protein and it might supply a basis for large‐scale production of the protein.

Identification and functional characterization of a novel fungal immunomodulatory protein from Postia placenta

“…The agglutinating activities of different FIPs toward different types of red blood cells changed to varying degrees (Bastiaan-Net et al, 2013;Li et al, 2012;Xue et al, 2008). However, none of the reported FIPs except FIPfve showed any hemagglutinating activity toward human red blood cells (Han et al, 2010;Ko et al, 1995;Li et al, 2012;Xue et al, 2008). In this study, rFIP-ppl presented moderate hemagglutinating activity on rabbit red blood cells but no agglutination in human red blood cells.…”

An aryl-alcohol oxidase of Pleurotus sapidus: heterologous expression, characterization, and application in a 2-enzyme system

“…In other studies, where C. cinerea was used as a host for recombinant production of foreign enzymes, no protein yields are given (Kikuchi et al 2004;Ogawa et al 1998;Muraguchi et al 2011). The expression of an immunomodulatory protein from Ganoderma sinense in C. cinerea reached 314 mg kg −1 of fresh mycelium (Han et al 2010). Expression yields of homologous laccases in C. cinereas can reach up to 115 mg L −1 (Rühl 2009).…”

“…A suitable basidiomycete which has been transformed for the first time almost 30 years ago (Binninger et al 1987) is the model organism Coprinopsis cinerea. Successful transformations of C. cinerea for heterologous expression of different proteins have been reported, e.g., for the green fluorescent protein GFP (Burns et al 2005), the immunomodulatory protein FIP-gsi (Han et al 2010), and different hydrolases (Kikuchi et al 2004;Cheng et al 2009). Heterologous production of oxidases in C. cinerea has only been described for two enzymes: for a peroxidase and a laccase (Ogawa et al 1998;Muraguchi et al 2011).…”

Fungal immunomodulatory proteins in the context of biomedicine

“…The FIPs can be also useful products for the treatment or prevention of autoimmune diseases and cancer, among others. Also to relieve pain or side effects caused by drugs in cancer patients (60)(61)70). For these reasons, the FIPs should be consider good candidates for new developments in the field of functional foods as well as in the pharmaceutical industry.…”