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Background  More than a dozen of fungal immunomodulatory proteins (FIPs) have been identified to date, most of which are from  Ganoderma  species. However, little is known about the similarities and differences between different  Ganoderma  FIPs’ bioactivities. In the current study, two FIP genes termed  FIP-gap1  and  FIP-gap2  from  G. applanatum , along with  LZ-8  and  FIP-gsi , another two representative  Ganoderma  FIP genes from  G. lucidum  and  G. sinense  were functionally expressed in  Pichia . Subsequently, bioactivities of four recombinant  Ganoderma  FIPs were demonstrated and compared.    Results  All the four  Ganoderma  FIP genes could be effectively expressed in  P. pastoris  GS115 at expression levels ranging from 197.5 to 264.3 mg L − 1  and simply purified by one step chromatography using HisTrap™ FF prepack columns. Amino acid sequence analysis showed that they all possessed the FIP conserved fragments. The homologies of different  Ganoderma  FIPs were from 72.6 to 86.4%. In vitro haemagglutination exhibited that FIP-gap1, FIP-gsi and LZ-8 could agglutinate human, sheep and mouse red blood cells but FIP-gap2 agglutinated none. Besides, the immunomodulation activities of these  Ganoderma  FIPs were as: rFIP-gap2 > rFIP-gap1 > rLZ-8 and rFIP-gsi in terms of proliferation stimulation and cytokine induction on murine splenocytes. Additionally, the cytotoxic activity of different FIPs was: rFIP-gap1 > rLZ-8 > rFIP-gsi > rFIP-gap2, examined by their inhibition of three human carcinomas A549, Hela and MCF-7.    Conclusions  Taken together, four typical  Ganoderma  FIP genes could be functionally expressed in  P. pastoris , which might supply as feasible efficient resources for further study and application. Both similarities and differences were indeed observed between  Ganoderma  FIPs in their amino acid sequences and bioactivities. Comprehensively, rFIP-gaps from  G. applanatum  proved to be more effective in immunomodulation and cytotoxic assays in vitro than rLZ-8 ( G. lucidum ) and rFIP-gsi ( G. sinense ).    Electronic supplementary material  The online version of this article (10.1186/s12896-018-0488-0) contains supplementary material, which is available to authorized users.

Recombinant Expression and Bioactivity Comparison of Four Typical Fungal Immunomodulatory Proteins from Three Main Ganoderma Species