Yes, there are at least tree reviews published about proteomic approaches to fungi. In spite of the used protocol depends of your own decision, I had used TCA/phenol with nice results.

Besides TCA/phenol, you may need to add some glass beads too.

Hi, you can check our video on fusarium at

http://www.jove.com/video/1690/toxin-induction-protein-extraction-from-fusarium-spp-cultures-for

or read a modified protocol at

http://onlinelibrary.wiley.com/doi/10.1002/elps.201200256/abstract.

good luck

Especially for surface-associated extracellular proteins you may wish to look at the protocols descibed in:

1. Comparative and functional genomics provide insights into the pathogenicity of dermatophytic fungi

Author(s): Burmester, A (Burmester, Anke)1,2; Shelest, E (Shelest, Ekaterina)3; Glockner, G (Gloeckner, Gernot)4; Heddergott, C (Heddergott, Christoph)1,2; Schindler, S (Schindler, Susann)5; Staib, P (Staib, Peter)6; Heidel, A (Heidel, Andrew)4; Felder, M (Felder, Marius)4,7; Petzold, A (Petzold, Andreas)4; Szafranski, K (Szafranski, Karol)4; Feuermann, M (Feuermann, Marc)8; Pedruzzi, I (Pedruzzi, Ivo)8; Priebe, S (Priebe, Steffen)3; Groth, M (Groth, Marco)4; Winkler, R (Winkler, Robert)9; Li, WJ (Li, Wenjun)10; Kniemeyer, O (Kniemeyer, Olaf)1; Schroeckh, V (Schroeckh, Volker)1; Hertweck, C (Hertweck, Christian)9; Hube, B (Hube, Bernhard)11; White, TC (White, Theodore C.)12; Platzer, M (Platzer, Matthias)4; Guthke, R (Guthke, Reinhard)3; Heitman, J (Heitman, Joseph)10; Wostemeyer, J (Woestemeyer, Johannes)2; Zipfel, PF (Zipfel, Peter F.)5; Monod, M (Monod, Michel)13; Brakhage, AA (Brakhage, Axel A.)1,2

Source: GENOME BIOLOGY Volume: 12 Issue: 1 Article Number: R7 DOI: 10.1186/gb-2011-12-1-r7 Published: 2011

2. ISOLATION AND ELECTROPHORETIC ANALYSIS OF SURFACE-PROTEINS OF THE ZYGOMYCETE ABSIDIA-GLAUCA

Author(s): TEEPE, H (TEEPE, H); BOTTGE, JA (BOTTGE, JA); WOSTEMEYER, J (WOSTEMEYER, J)

The protocol works nicely for several fungu from ascomycetes and zygomycetes.

Good luck, nice project! - Johannes W.

Just some comments on your question. I would suggest before you even rush onto any method, know the physiology of the fungi you are handling first. Then, it will be nice to determine whether the fungi excrete the protein of you interest in the cultivation media in high amount of not. You can check this by merely determining the protein concentration using the crude extract first. Once you have a hint on the excretion levels and the nature of your protean, you can then optimize the appropiete methods available to maximize the yields. If you want your protein in pure form and active, there are several kits for purification which you can use. I hope this can guide you anyway.

Extracellular proteins you can get if you grow the fungy in liquid synthetic medium (for example Chapek medium). In this case you can delete cells by centrifugation and use the cultural liquid - lyophilize it and use further. Extraction of total proteins you can do from the pellet of cells by 1% SDS at 40-50 degrees during 10-20 min.

As already mentioned, TCA/phenol with glass beads method will solve your problem.

Some of these answers are all over the place. The video linked by Matias is quite good. Very detailed and similar to things that I have done as well. The link from Brakahage from Johannes is likely good as well. It is simply not true that you can't break fungal cells with glass beads. I've done that quite a bit. Also, most of the methods here include SDS and TCA precipitation. Good methods, but not if you want to recover native proteins or retain activity. It may be difficult to obtain Postia that is 'growing on wood' as you state. The method used for the Postia genome paper came from the Forest Products Lab, published here:

http://www.fpl.fs.fed.us/documnts/pdf2006/fpl_2006_wymelenberg002.pdf

I used phenol chloroform protocol modified to break the wall in the demateaceous fungus check http://www.ncbi.nlm.nih.gov/pubmed/22734968

Thank you so much everyone for all you comments!

It was a very useful questionaire and answers. I too have worked on fungal proteins but exclusively on extracellular. Extracellular proteins from filamentous fungi were easily precipitated from the production medium by 80% acetone in our case.

You can look up several papers published from our for fungal protein (both mycelial and secreted into media) extraction and electrophoresis in this regard.

Yajima and Kav 2006. Proteomics 6: 5995

Liang et al. 2010 Fungal Biology 114:619

Liang et al. 2010. Journal of Proteome Res 9:3290

If you can not access these publications, I can send you the pdf files.

Hope this helps.

Thank you to all friends