title: Extracellular expression and efficient purification of a functional recombinant Volvariella volvacea immunomodulatory protein (FIP-vvo) using Pichia pastoris system.
abstract: The fungal immunomodulatory proteins (FIPs) are a new protein family identified from several edible and medical mushrooms and play an important role in antitumor, anti-allergy and immunomodulating activities. A gene encoding the FIP-vvo was cloned from the mycelia of Volvariella volvacea and recombinant expressed in the Pichia pastoris expression system. SDS-PAGE, amino acid composition and circular dichroism analyses of the recombinant FIP-vvo (reFIP-vvo) indicated that the gene was correctly and successfully expressed. In vitro assays of biological activities revealed that the reFIP-vvo exhibited similar immunomodulating capacities as native form. The reFIP-vvo significantly stimulated the proliferation of mouse spleen lymphocytes and apparently enhanced the expression level of IFN-γ released from the mouse splenocytes. Taken together, the FIP-vvo gene from V. volvacea has been integrated into the yeast genome and expressed effectively at a high level (about 410mg/L), it was capable of agglutinating sheep and rat red blood cells. The reFIP-vvo possessed very similar biological activities to native FIPs, suggesting its potential application as a food supplement or immunomodulating agent in pharmaceuticals and even medical studies.
authors.affiliation: Institute of Frontier Medical Science of Jilin University, Changchun City 130021, China.
authors.forename: Xilin
authors.initials: X
authors.lastname: Sun
authors.affiliation: Institute of Frontier Medical Science of Jilin University, Changchun City 130021, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Huang
authors.affiliation: Institute of Frontier Medical Science of Jilin University, Changchun City 130021, China.
authors.forename: Sijia
authors.initials: S
authors.lastname: Xiao
authors.affiliation: Institute of Frontier Medical Science of Jilin University, Changchun City 130021, China.
authors.forename: Chongyang
authors.initials: C
authors.lastname: Liang
authors.affiliation: Institute of Frontier Medical Science of Jilin University, Changchun City 130021, China.
authors.forename: Shuqin
authors.initials: S
authors.lastname: Zhang
authors.affiliation: Institute of Frontier Medical Science of Jilin University, Changchun City 130021, China.
authors.forename: Zhiyi
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Institute of Frontier Medical Science of Jilin University, Changchun City 130021, China. Electronic address: s.fei1958@gmail.com.
authors.forename: Fei
authors.initials: F
authors.lastname: Sun
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D049109:Cell Proliferation / Q000187:drug effects; D002942:Circular Dichroism; D005656:Fungal Proteins / Q000096:biosynthesis* / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology; D015966:Gene Expression Regulation, Fungal; D056747:Immunomodulation / Q000187:drug effects / Q000235:genetics*; D008214:Lymphocytes / Q000166:cytology / Q000187:drug effects; D051379:Mice; D010843:Pichia; D013154:Spleen / Q000166:cytology / Q000187:drug effects; D055367:Volvariella / Q000737:chemistry / Q000235:genetics*
chemical_list: D005656:Fungal Proteins
keywords: Immunomodulatory; Pichia pastoris; Volvariella volvacea; reFIP-vvo