Evidence map›Paper›PMID 39184091›Full record

ArticleResearch square2024

SANS investigation of fungal loosenins reveal substrate dependent impacts of protein 1 action on inter-fibril distance and packing order of cellulosic substrates.

Deepika Dahiya, Zsuzsanna Péter-Szabó, Manjula Senanayake, Sai Venkatesh Pingali, Wellington C Leite, James Byrnes, Garry W Buchko, Pramod Sivan, Francisco Vilaplana, Emma Master and 1 more

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Article in Research square, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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1 · What the graph read from it

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4 · The record

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5 · Who and what money

Authors and funding

11 authors.

Deepika Dahiya
Zsuzsanna Péter-Szabó
Manjula Senanayake
Sai Venkatesh Pingali
Wellington C Leite
James Byrnes
Garry W Buchko
Pramod Sivan
Francisco Vilaplana
Emma Master
Hugh O'Neill

Funding

X-ray Scattering Technology CoreP30GM133893 · NIGMS · BROOKHAVEN SCIENCE ASSOC-BROOKHAVEN LAB · PI Sean McSweeney · 2019 to 2026
$38.6M
A Simultaneous SAXS/WAXS Detector System for Solving Biological StructuresS10OD012331 · OD · STATE UNIVERSITY NEW YORK STONY BROOK · PI ALLAIRE, MARC · 2012 to 2012
$1.1M
NIGMS NIH HHS P30 GM133893NIH HHS S10 OD012331
6 · The paper itself

Abstract

backgroundMicrobial expansin-related proteins include fungal loosenins, which have been previously shown to disrupt cellulose networks and enhance the enzymatic conversion of cellulosic substrates. Despite showing beneficial impacts to cellulose processing, detailed characterization of cellulosic materials after loosenin treatment is lacking. In this study, small-angle neutron scattering (SANS) was used to investigate the effects of three recombinantly produced loosenins that originate from Phanerochaete carnosa, PcaLOOL7, PcaLOOL9, and PcaLOOL12, on the organization of holocellulose preparations from Eucalyptus and Spruce wood samples.

resultsWhereas the SANS analysis of Spruce holocellulose revealed an increase in interfibril spacing of neighboring cellulose microfibrils following treatment with PcaLOOL12 and to a lesser extent PcaLOOL7, the analysis of Eucalyptus holocellulose revealed a reduction in packing number following treatment with PcaLOOL12 and to a lesser extent PcaLOOL9. Parallel SEC-SAXS characterization of PcaLOOL7, PcaLOOL9, and PcaLOOL12 indicated the proteins likely function as monomers; moreover, all appear to retain a flexible disordered N-terminus and folded C-terminal region. The comparatively high impact of PcaLOOL12 motivated its NMR structural characterization, revealing a double-psi b-barrel (DPBB) domain surrounded by three alpha-helices - the largest nestled against the DPBB core and the other two part of loops extending from the core.

conclusionsThe SANS analysis of PcaLOOL action on holocellulose samples confirms their ability to disrupt cellulose fiber networks and suggests a progression from reducing microfibril packing to increasing interfibril distance. The most impactful PcaLOOL, PcaLOOL12, was previously observed to be the most highly expressed loosenin in P. carnosa. Its structural characterization herein reveals its stabilization through two disulfide linkages, and an extended N-terminal region distal to a negatively charged and surface accessible polysaccharide binding groove.

Identifiers

PMID39184091
PMCPMC11343303

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