Analysis of the Structure and Conformational States of DewA Gives Insight into the Assembly of the Fungal Hydrophobins

Morris, Vanessa K.; Kwan, Ann H.; Sunde, Margaret

doi:10.1016/j.jmb.2012.10.021

Cited by 52 publications

(66 citation statements)

References 30 publications

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“…The recombinant production of HFBs has been achieved in filamentous fungi (46,47), yeasts (Pichia pastoris [48]) and bacteria (E. coli [49]). The latter case often leads to HFB production in intracellular inclusion bodies, thus warranting additional unfolding/refolding steps.…”

Section: Discussionmentioning

confidence: 99%

Two Novel Class II Hydrophobins from Trichoderma spp. Stimulate Enzymatic Hydrolysis of Poly(Ethylene Terephthalate) when Expressed as Fusion Proteins

Espino-Rammer

Ribitsch

Przylucka

et al. 2013

Appl Environ Microbiol

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Poly(ethylene terephthalate) (PET) can be functionalized and/or recycled via hydrolysis by microbial cutinases. The rate of hydrolysis is however low. Here, we tested whether hydrophobins (HFBs), small secreted fungal proteins containing eight positionally conserved cysteine residues, are able to enhance the rate of enzymatic hydrolysis of PET. Species of the fungal genus Trichoderma have the most proliferated arsenal of class II hydrophobin-encoding genes among fungi. To this end, we studied two novel class II HFBs (HFB4 and HFB7) of Trichoderma. HFB4 and HFB7, produced in Escherichia coli as fusions to the C terminus of glutathione S-transferase, exhibited subtle structural differences reflected in hydrophobicity plots that correlated with unequal hydrophobicity and hydrophily, respectively, of particular amino acid residues. Both proteins exhibited a dosage-dependent stimulation effect on PET hydrolysis by cutinase from Humicola insolens, with HFB4 displaying an adsorption isotherm-like behavior, whereas HFB7 was active only at very low concentrations and was inhibitory at higher concentrations. We conclude that class II HFBs can stimulate the activity of cutinases on PET, but individual HFBs can display different properties. The present findings suggest that hydrophobins can be used in the enzymatic hydrolysis of aromatic-aliphatic polyesters such as PET. P oly(ethylene terephthalate) (PET) is a thermoplastic polyester with excellent tensile and impact strength, transparency, and appropriate thermal stability (1). Because of its high production (36 million tons per year [2]), PET constitutes a significant waste material, which, although not representing a direct hazard to the environment, is not readily decomposed in nature.Enzymatic recycling of polymers and particularly of PET basically would break down the polymer into its building blocks ethylene glycol (EG) and terephthalic acid (TA). These have a high value and can be reused in chemical synthesis, including the production of PET. This would avoid current limitations in PET recycling, which, e.g., requires pure PET fractions or has to fight with enrichment of contaminants (3). In addition, the surface modification of PET to increase its hydrophilicity is an essential step in processing for many applications ranging from textiles to medical and electronics. Synthetic polymers and particularly PET show excellent chemical resistance, but this feature makes them very difficult to be functionalized and is a great drawback for the processing steps. Current methods, including the use of harsh chemicals, concentrated acid or alkali, or different types of plasma approaches pursue to insert reactive moieties. Therefore, enzymatic strategies have recently received considerable attention (for a review, see reference 2) due to their environmentally friendly profile compared to currently used techniques such as those based on concentrated alkali (4) or the limitation of plasma methods (5) to planar surfaces.Compared to traditional approaches, enzymatic partial hydrol...

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Section: Discussionmentioning

confidence: 99%

Two Novel Class II Hydrophobins from Trichoderma spp. Stimulate Enzymatic Hydrolysis of Poly(Ethylene Terephthalate) when Expressed as Fusion Proteins

Espino-Rammer

Ribitsch

Przylucka

et al. 2013

Appl Environ Microbiol

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“…I-TASSER was used to predict a 3D structure for T . vaccinum hydrophobins [57] using DewA [58], Hfb2 [59], and EAS [60] as templates with HADDOCK driven prediction to model protein-protein interactions (F86 = active; S85-G89 = passive or F54 = active; V52-D55 = passive; solvent = water [23, 61]). Protein calculator (Innovagen AB, Sweden) was used for the prediction of pH net charge.…”

Section: Methodsmentioning

confidence: 99%

Hydrophobins in the Life Cycle of the Ectomycorrhizal Basidiomycete Tricholoma vaccinum

et al. 2016

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Hydrophobins—secreted small cysteine-rich, amphipathic proteins—foster interactions of fungal hyphae with hydrophobic surfaces, and are involved in the formation of aerial hyphae. Phylogenetic analyses of Tricholoma vaccinum hydrophobins showed a grouping with hydrophobins of other ectomycorrhizal fungi, which might be a result of co-evolution. Further analyses indicate angiosperms as likely host trees for the last common ancestor of the genus Tricholoma. The nine hydrophobin genes in the T. vaccinum genome were investigated to infer their individual roles in different stages of the life cycle, host interaction, asexual and sexual development, and with respect to different stresses. In aerial mycelium, hyd8 was up-regulated. In silico analysis predicted three packing arrangements, i.e., ring-like, plus-like and sheet-like structure for Hyd8; the first two may assemble to rodlets of hydrophobin covering aerial hyphae, whereas the third is expected to be involved in forming a two-dimensional network of hydrophobins. Metal stress induced hydrophobin gene hyd5. In early steps of mycorrhization, induction of hyd4 and hyd5 by plant root exudates and root volatiles could be shown, followed by hyd5 up-regulation during formation of mantle, Hartig’ net, and rhizomorphs with concomitant repression of hyd8 and hyd9. During fruiting body formation, mainly hyd3, but also hyd8 were induced. Host preference between the compatible host Picea abies and the low compatibility host Pinus sylvestris could be linked to a stronger induction of hyd4 and hyd5 by the preferred host and a stronger repression of hyd8, whereas the repression of hyd9 was comparable between the two hosts.

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“…While hydrophobins from both phyla have been characterized, high-resolution structures exist only for those of ascomycota origin: the Class I EAS, DewA, MPG1, and Class II NC2, HFBI, and HFBII81920212223. To address whether basidiomycota hydrophobins share structural and functional features with these proteins, we preliminarily characterized SC16 ( hyd1 ) from Schizophyllum commune and determined its structure.…”

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confidence: 99%

Characterization of a Basidiomycota hydrophobin reveals the structural basis for a high-similarity Class I subdivision

Gandier

Langelaan

Won

et al. 2017

Sci Rep

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Class I hydrophobins are functional amyloids secreted by fungi. They self-assemble into organized films at interfaces producing structures that include cellular adhesion points and hydrophobic coatings. Here, we present the first structure and solution properties of a unique Class I protein sequence of Basidiomycota origin: the Schizophyllum commune hydrophobin SC16 (hyd1). While the core β-barrel structure and disulphide bridging characteristic of the hydrophobin family are conserved, its surface properties and secondary structure elements are reminiscent of both Class I and II hydrophobins. Sequence analyses of hydrophobins from 215 fungal species suggest this structure is largely applicable to a high-identity Basidiomycota Class I subdivision (IB). To validate this prediction, structural analysis of a comparatively distinct Class IB sequence from a different fungal order, namely the Phanerochaete carnosa PcaHyd1, indicates secondary structure properties similar to that of SC16. Together, these results form an experimental basis for a high-identity Class I subdivision and contribute to our understanding of functional amyloid formation.

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Analysis of the Structure and Conformational States of DewA Gives Insight into the Assembly of the Fungal Hydrophobins

Cited by 52 publications

References 30 publications

Two Novel Class II Hydrophobins from Trichoderma spp. Stimulate Enzymatic Hydrolysis of Poly(Ethylene Terephthalate) when Expressed as Fusion Proteins

Two Novel Class II Hydrophobins from Trichoderma spp. Stimulate Enzymatic Hydrolysis of Poly(Ethylene Terephthalate) when Expressed as Fusion Proteins

Hydrophobins in the Life Cycle of the Ectomycorrhizal Basidiomycete Tricholoma vaccinum

Characterization of a Basidiomycota hydrophobin reveals the structural basis for a high-similarity Class I subdivision

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