Saturday, 26 November 2016

Marine fungi as a source of new hydrophobins?

Fungi are a monophyletic group that has been found across all parts of the world. These fungi are very diverse, some being filamentous and containing structures known as hyphae, whilst other fungi are single celled with no hyphae. Many of these filamentous fungi produce hydrophobins - small proteins of about 100 amino acids that are the most powerful surface-active proteins to our current knowledge. Hydrophobins have an amphiphilic structure, with their hydrophobic aliphatic side chains able to create a coating on the surface of an object. These properties have been shown to have uses in coating of spores, which allows the fungus to move about and to attach to different surfaces.

Cicatiello et al (2016) aimed to identify marine fungi as a source of hydrophobins. Hydrophobins were split into Class I and Class II, depending on their properties. Class I form highly stable, highly insoluble aggregates and have distinct rodlets, whereas Class II are less stable and lack the ability to form rodlets.

Marine fungi was isolated from the seagrass Posidonia oceanica, the green alga Flabellia petiolata and the brown alga Padina pavonica near Elba island in the Mediterranean sea. Each marine fungal strain was maintained on an agar plate at 20OC. To extract Class I and Class II hydrophobins from the culture broth, proteins were aggregated by bubbling air using a Waring blend, and the foam was collected and treated with 20% trichloroacetic acid.

To extract Class II hydrophobins from the mycelium, mycelia were washed with water and proteins were extracted using 60% ethanol in a bath sonicator. Class I hydrophobins were extracted using 2% sodium dodecyl sulphate, water, 60% ethanol and trifluoroacetic acid.

The results showed that 23 out of the 100 strains of marine fungi were chosen due to their foam producing capabilities in shaken cultures, thus showing the production of biosurfactants. The isolation of Class I and Class II hydrophobins from the culture broth and the mycelium allowed the identification of 6 new putative hydrophobins that could be used in biotechnological instances in the future.

This study provided evidence for marine fungi as sources of hydrophobins, and seemed to go beyond the aims they set by trying to look at the functions of the identified hydrophobins. Whilst this is a good thing, it did seem to get confusing at times as to what they were trying to achieve. The way it was written was also confusing at times as the parts of the methods seemed to appear in the results and discussion sections. This paper could be more clear and concise on just what it was trying to show, and could be written better, but I think the overall conclusion and results seemed to show that the study was successful in reaching its aim.


Reviewed paper: Cicatiello, Paola., Gravagnuolo, Alfredo. Maria., Gnavi, Giorgio., Varese, Giovanna. Cristina., and Giardina, Paola. (2016). Marine fungi as a source of new hydrophobins. International journal of biological macromolecules. 92: 1229-1233. http://www.sciencedirect.com/science/article/pii/S0141813016311928  

4 comments:

  1. Hi Amy,
    Great post, you mentioned that the authors aimed to look at the function of the hydrophobins, did they conclusively determine any of the functions?
    Do you think that the marine fungal hydrophobins seen in this study are widespread in marine fungi as they are in terrestrial species?
    Thanks
    Natasha

    ReplyDelete
  2. Hi Natasha,

    Thank you for your questions. They showed the ability for the putative hydrophobins to self-assemble into a stable amphiphilic layer, and to functionalize a solid surface. This helped them to determine which hydrophobins were in Class I and in Class II. The emulsification capacity was also tested in the 6 new putative hydrophobins, and all 6 were able to produce oil/water emulsions.
    I think that it is highly possible that these hydrophobins are widespread in marine fungi, as being able to create surfactants seems to be incredibly useful, especially in the marine environment where the fungi needs to move through the liquid medium.

    I hope this answers your questions.

    Thanks,
    Amy

    ReplyDelete
  3. Hi Amy,

    Love this sort of paper, looking into possible biotechnological uses. I was wondering though, is the Class I and Class II system universal or just used by the authors?

    Along with possible biotech uses, could these Hydrophobins be possible used for bioremediation purposes as I have seen it mentioned that many biosurfactants also enhanced emulsification with biodegradation potential of hydrocarbon pollutants.

    Yadav, A., Manna, S., Pandiyan, K., Singh, A., Kumar, M., Chakdar, H., Kashyap, P. and Srivastava, A. (2016). Isolation and characterization of biosurfactant producing Bacillus sp. from diesel fuel-contaminated site. Microbiology, 85(1), pp.56-62.

    ReplyDelete
    Replies
    1. Hi Stefan,

      Thank you for your question. It looks to me as though this system is universal as it seems to be mentioned in other papers (e.g. Askolin et al, 2006).

      You've raised an interesting point that I hadn't actually considered. In a paper by Hobley et al, 2013, it's shown that biofilms have critical roles in bioremediation, and assembles with the help of a small secreted protein known as BslA. BslA is a structurally defined hydrophobin, so I suppose you could say that hydrophobins are used in bioremediation as this is evidence, and I expect there will be more evidence if there isn't already.

      I hope this answers your question.

      Thanks,
      Amy

      Askolin, S., Linder, M., Scholtmeijer, K., Tenkanen, M., Penttila, M., de Vocht, M. L., and Wosten, H. A. (2006). Interaction and comparison of a class I hydrophobin from Schizophyllum commune and class II hydrophobins from Trichoderma reesei. Biomacromolecules. 7(4): 1295-1301. https://www.ncbi.nlm.nih.gov/pubmed/16602752

      Hobley, L., Ostrowski, A., Rao, F. V., Bromley, K. M., Porter, M., Prescott, A. R., MacPhee, C. E., van Aalten, D. M. F. and Stanley-Wall, N. R. (2013). BslA is a self-assembling bacterial hydrophobin that coats the Bacillus subtilis biofilm. PNAS. 110(33): 13600-13605. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3746881/

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