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Expression of recombinant human interleukin-32 in Pleurotus eryngii

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Abstract

Interleukin-32 is a novel human cytokine implicated in several inflammatory and autoimmune diseases. Recombinant interleukin -32 can be produced and found to be useful in many fields. Until now, there has been no report on recombinant hIL-32 expressed in basidomycete fungus, Pleurotus eryngii. In this study, we examined whether the popular edible mushroom Pleurotus eryngii could support the expression of novel protein hIL-32. A binary vector pCAMBIA1304 containing the hIL-32 gene was constructed and introduced into Pleurotus eryngii via Agrobacterium tumefaciens-mediated transformation. The expression of hIL-32 was confirmed by PCR, Southern blot and western blot analysis. The recombinant hIL-32 reached a maximum expression level of 1.9 % of total soluble protein in transgenic mycelia. These results suggest that Pleurotus eryngii expression system can be effective for the production of rhIL-32 at an economically relevant level.

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References

  • Berends E, Scholtmeijer K, Wosten HA, Bosch D, Lugones LG (2009) The use of mushroom-forming fungi for the production of N-glycosylated therapeutic proteins. Trends Microbiol 17:439–443

    Article  PubMed  CAS  Google Scholar 

  • Bundock P, Hooykaas PJJ (1996) Integration of Agrobacterium tumefaciens T-DNA in the Saccharomyces cerevisiae genome by illegitimate recombination. Proc Natl Acad Sci USA 93:15272–15275

    Article  PubMed  CAS  Google Scholar 

  • Bundock PA, Den Dulk-Ras A, Beijersbergen A, Hooykaas PJJ (1995) Trans-kingdom T-DNA transfer from Agrobacterium tumefaciens to Saccharomyces cerevisiae. EMBO J 14:3206–3214

    PubMed  CAS  Google Scholar 

  • Chen X, Stone M, Schlagnhaufer C, Romaine CP (2000) A fruiting body tissue method for efficient Agrobacterium-mediated transformation of Agaricus bisporus. Appl Environ Microbiol 66:4510–4513

    Article  PubMed  CAS  Google Scholar 

  • Chen Q, Carroll HP, Gadina M (2006) The newest interleukins: recent additions to the ever-growing cytokine family. Vitam Horm 74:207–228

    Article  PubMed  CAS  Google Scholar 

  • DeGroot MJA, Bundock P, Hooykaas PJJ, Beijersbergen AGM (1998) Agrobacterium tumefaciens-mediated transformation of filamentous fungi. Nat Biotechnol 16:839–842

    Article  CAS  Google Scholar 

  • Demain AL, Vaishnav P (2009) Production of recombinant proteins by microbes and higher organisms. Biotechnol Adv 27:297–306

    Article  PubMed  CAS  Google Scholar 

  • Fernandez JM, Hoeffler JP (1999) Gene expression systems. Academic, San Diego

    Google Scholar 

  • Goda C, Kanaji T, Kanaji S, Tanaka G, Arima K, Ohno S, Izuhara K (2006) Involvement of IL-32 in activation-induced cell death in T cells. Int Immunol 18:233–240

    Article  PubMed  CAS  Google Scholar 

  • Holsters M, De Waele D, Depicker A, Messens E, van Montague, Schell J (1978) Transfection and transformation of Agrobacterium tumefaciens. Mol Gen Genet 163:168–181

    Google Scholar 

  • Joosten LA, Netea MG, Kim SH, Yoon DY, Oppers-Walgreen B, Radstake TR, Barrera P, van de Loo FA, Dinarello CA, van den Berg WB (2006) IL-32, a proinflammatory cytokine in rheumatoid arthritis. Proc Natl Acad Sci USA 103:3298–3303

    Article  PubMed  CAS  Google Scholar 

  • Kim SH, Han SY, Azam T, Yoon DY, Dinarello CA (2005) Interleukin-32: a cytokine and inducer of TNF alpha. Immunity 22:131–142

    PubMed  CAS  Google Scholar 

  • Kim KH, Shim JH, Seo EH, Cho MC, Kang JW, Kim SH, Yu DY, Song EY, Lee HG, Sohn JH, Kim J, Dinarello CA, Yoon DY (2008) Interleukin-32 monoclonal antibodies for immunohistochemistry, Western blotting, and ELISA. J Immunol Meth 333:38–50

    Article  CAS  Google Scholar 

  • Kim S, Sapkota K, Choi BS, Kim SJ (2010) Expression of human growth hormone gene in Pleurotus eryngii. Cent Eur J Biol (In press) doi:10.2478/s11535-010-0071-8 Online First™

  • Kobayashi H, Lin PC (2009) Molecular characterization of IL-32 in human endothelial cells. Cytokine 46:351–358

    Article  PubMed  CAS  Google Scholar 

  • Kobayashi H, Huang J, Ye F, Shyr Y, Blackwell TS, Lin PC (2010) Interleukin-32beta propagates vascular inflammation and exacerbates sepsis in a mouse model. PLoS One 5:e9458

    Article  PubMed  Google Scholar 

  • Li W, Liu Y, Mukhtar MM, Gong R, Pan Y, Rasool ST, Gao Y, Kang L, Hao Q, Peng G, Chen X, Wu J, Zhu Y (2008) Activation of interleukin-32 pro-inflammatory pathway in response to influenza A virus infection. PLoS ONE 3:e1985

    Article  PubMed  Google Scholar 

  • Netea MG, Azam T, Ferwerda G, Girardin SE, Walsh M, Park JS, Abraham E, Kim JM, Yoon DY, Dinarello CA, Kim SH (2005) IL-32 synergizes with nucleotide oligomerization domain (NOD) 1 and NOD2 ligands for IL- 1beta and IL-6 production through a caspase 1-dependent mechanism. Proc Natl Acad Sci USA 102:16309–16314

    Article  PubMed  CAS  Google Scholar 

  • Netea MG, Lewis EC, Azam T, Joosten LA, Jaekal J, Bae SY, Dinarello CA, Kim SH (2008) Interleukin-32 induces the differentiation of monocytes into macrophage-like cells. Proc Natl Acad Sci USA 9:3515–3520

    Article  Google Scholar 

  • Nevalainen KM, Teo VS, Bergquist PL (2005) Heterologous protein expression in filamentous fungi. Trends Biotechnol 23:468–474

    Article  PubMed  CAS  Google Scholar 

  • Nold MF, Nold-Petry CA, Pott GB, Zepp JA, Saavedra MT, Kim SH, Dinarello CA (2008) Endogenous IL-32 controls cytokine and HIV-1 production. J Immunol 181:557–565

    PubMed  CAS  Google Scholar 

  • Punt PJ, van Biezen N, Conesa A, Albers A, Mangnus J, van den Hondel C (2002) Filamentous fungi as cell factories for heterologous protein production. Trends Biotechnol 5:200–206

    Article  Google Scholar 

  • Rodriguez Estrada AE, Royse DJ (2007) Yield, size and bacterial blotch resistance of Pleurotus eryngii grown on cottonseed hulls/oak sawdust supplemented with manganese, copper and whole ground soybean. Bioresour Technol 98:1898–1906

    Article  PubMed  CAS  Google Scholar 

  • Royse DJ (1999) Yield stimulation of king oyster mushroom, Pleurotus eryngii, by brewer’s grain and SpawnMate IISE®supplementation of cottonseed hull and woodchip substrate. Mushroom News 47(2):4–8

    Google Scholar 

  • Scott OR, Bendich AJ (1994) Extraction of total cellular DNA from plant, algae and fungi. In: Gelvin SB, Schilperoort RA (eds) Plant molecular biology manual, D1. Kluwer, Dordercht, pp 1–8

    Google Scholar 

  • Shioya M, Nishida A, Yagi Y, Ogawa A, Tsujikawa T, Kim-Mitsuyama S, Takayanagi A, Shimizu N, Fujiyama Y, Andoh A (2007) Epithelial overexpression of interleukin- 32alpha in inflammatory bowel disease. Clin Exp Immunol 149:480–486

    Article  PubMed  CAS  Google Scholar 

  • Shoda H, Fujio K, Yamaguchi Y, Okamoto A, Sawada T, Kochi Y, Yamamoto K (2006) Interactions between IL-32 and tumor necrosis factor alpha contribute to the exacerbation of immune-inflammatory diseases. Arthritis Res Ther 8:R166

    Article  PubMed  Google Scholar 

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Acknowledgement

This study was supported by Technology Development Program of the Ministry of Agriculture and Forestry (ARPC), Republic of Korea, and was conducted with research funds from Gwangju University in 2010.

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Authors and Affiliations

  1. Department of Biotechnology, Chosun University, 375 Seosuk-dong, Dong-gu, Gwangju, 501-759, Republic of Korea

    Sang Jun Chung, Kumar Sapkota, Bong-Suk Choi & Sung-Jun Kim

  2. Department of Alternative Medicine, Gwangju University, Gwangju, 503-703, Republic of Korea

    Seung Kim

  3. Department of Sport and Leisure Studies, Gwangju University, Gwangju, 503-703, Republic of Korea

    Changho Shin

  4. Central Department of Zoology, Tribhuvan University, Kirtipur, Kathmandu, Nepal

    Kumar Sapkota

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  1. Sang Jun Chung
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  2. Seung Kim
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  3. Kumar Sapkota
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  4. Bong-Suk Choi
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  5. Changho Shin
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  6. Sung-Jun Kim
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Correspondence to Sung-Jun Kim.

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Sang Jun Chung and Seung Kim contributed equally to this study

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Chung, S.J., Kim, S., Sapkota, K. et al. Expression of recombinant human interleukin-32 in Pleurotus eryngii . Ann Microbiol 61, 331–338 (2011). https://doi.org/10.1007/s13213-010-0146-9

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  • DOI: https://doi.org/10.1007/s13213-010-0146-9

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