Functional and Structural Characterizations of Lytic Polysaccharide Monooxygenase, Which Cooperates Synergistically with Cellulases, from Ceriporiopsis subvermispora

2022 ◽  
Author(s):  
Huyen Nguyen ◽  
Keiko Kondo ◽  
Yusei Yagi ◽  
Yu Iseki ◽  
Nagi Okuoka ◽  
...  
Keyword(s):  
Lytic Polysaccharide Monooxygenase ◽  
Ceriporiopsis Subvermispora ◽  
Structural Characterizations ◽  
Polysaccharide Monooxygenase

scholarly journals Temporal Alterations in the Secretome of the Selective Ligninolytic Fungus Ceriporiopsis subvermispora during Growth on Aspen Wood Reveal This Organism's Strategy for Degrading Lignocellulose

2014 ◽  
Vol 80 (7) ◽  
pp. 2062-2070 ◽  
Author(s):  
Chiaki Hori ◽  
Jill Gaskell ◽  
Kiyohiko Igarashi ◽  
Phil Kersten ◽  
Michael Mozuch ◽  
...  
Keyword(s):  
Cellulose Degradation ◽  
Alcohol Oxidase ◽  
White Rot ◽  
Protein Abundance ◽  
Lytic Polysaccharide Monooxygenase ◽  
Content Type ◽  
Ceriporiopsis Subvermispora ◽  
Liquid Chromatography Tandem Mass ◽  
Nano Liquid Chromatography ◽  
Polysaccharide Monooxygenase

ABSTRACTThe white-rot basidiomycetes efficiently degrade all wood cell wall polymers. Generally, these fungi simultaneously degrade cellulose and lignin, but certain organisms, such asCeriporiopsis subvermispora, selectively remove lignin in advance of cellulose degradation. However, relatively little is known about the mechanism of selective ligninolysis. To address this issue,C. subvermisporawas grown in liquid medium containing ball-milled aspen, and nano-liquid chromatography-tandem mass spectrometry was used to identify and estimate extracellular protein abundance over time. Several manganese peroxidases and an aryl alcohol oxidase, both associated with lignin degradation, were identified after 3 days of incubation. A glycoside hydrolase (GH) family 51 arabinofuranosidase was also identified after 3 days but then successively decreased in later samples. Several enzymes related to cellulose and xylan degradation, such as GH10 endoxylanase, GH5_5 endoglucanase, and GH7 cellobiohydrolase, were detected after 5 days. Peptides corresponding to potential cellulose-degrading enzymes GH12, GH45, lytic polysaccharide monooxygenase, and cellobiose dehydrogenase were most abundant after 7 days. This sequential production of enzymes provides a mechanism consistent with selective ligninolysis byC. subvermispora.

scholarly journals A Lytic Polysaccharide Monooxygenase with Broad Xyloglucan Specificity from the Brown-Rot Fungus Gloeophyllum trabeum and Its Action on Cellulose-Xyloglucan Complexes

2016 ◽  
Vol 82 (22) ◽  
pp. 6557-6572 ◽  
Author(s):  
Yuka Kojima ◽  
Anikó Várnai ◽  
Takuya Ishida ◽  
Naoki Sunagawa ◽  
Dejan M. Petrovic ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Dynamic Viscosity ◽  
Brown Rot ◽  
Primary Cell Wall ◽  
Gloeophyllum Trabeum ◽  
Lytic Polysaccharide Monooxygenase ◽  
Content Type ◽  
Brown Rot Fungus ◽  
Sensitive Method ◽  
Polysaccharide Monooxygenase

ABSTRACTFungi secrete a set of glycoside hydrolases and lytic polysaccharide monooxygenases (LPMOs) to degrade plant polysaccharides. Brown-rot fungi, such asGloeophyllum trabeum, tend to have few LPMOs, and information on these enzymes is scarce. The genome ofG. trabeumencodes four auxiliary activity 9 (AA9) LPMOs (GtLPMO9s), whose coding sequences were amplified from cDNA. Due to alternative splicing, two variants ofGtLPMO9A seem to be produced, a single-domain variant,GtLPMO9A-1, and a longer variant,GtLPMO9A-2, which contains a C-terminal domain comprising approximately 55 residues without a predicted function. We have overexpressed the phylogenetically distinctGtLPMO9A-2 inPichia pastorisand investigated its properties. Standard analyses using high-performance anion-exchange chromatography–pulsed amperometric detection (HPAEC-PAD) and mass spectrometry (MS) showed thatGtLPMO9A-2 is active on cellulose, carboxymethyl cellulose, and xyloglucan. Importantly, compared to other known xyloglucan-active LPMOs,GtLPMO9A-2 has broad specificity, cleaving at any position along the β-glucan backbone of xyloglucan, regardless of substitutions. Using dynamic viscosity measurements to compare the hemicellulolytic action ofGtLPMO9A-2 to that of a well-characterized hemicellulolytic LPMO,NcLPMO9C fromNeurospora crassarevealed thatGtLPMO9A-2 is more efficient in depolymerizing xyloglucan. These measurements also revealed minor activity on glucomannan that could not be detected by the analysis of soluble products by HPAEC-PAD and MS and that was lower than the activity ofNcLPMO9C. Experiments with copolymeric substrates showed an inhibitory effect of hemicellulose coating on cellulolytic LPMO activity and did not reveal additional activities ofGtLPMO9A-2. These results provide insight into the LPMO potential ofG. trabeumand provide a novel sensitive method, a measurement of dynamic viscosity, for monitoring LPMO activity.IMPORTANCECurrently, there are only a few methods available to analyze end products of lytic polysaccharide monooxygenase (LPMO) activity, the most common ones being liquid chromatography and mass spectrometry. Here, we present an alternative and sensitive method based on measurement of dynamic viscosity for real-time continuous monitoring of LPMO activity in the presence of water-soluble hemicelluloses, such as xyloglucan. We have used both these novel and existing analytical methods to characterize a xyloglucan-active LPMO from a brown-rot fungus. This enzyme,GtLPMO9A-2, differs from previously characterized LPMOs in having broad substrate specificity, enabling almost random cleavage of the xyloglucan backbone.GtLPMO9A-2 acts preferentially on free xyloglucan, suggesting a preference for xyloglucan chains that tether cellulose fibers together. The xyloglucan-degrading potential ofGtLPMO9A-2 suggests a role in decreasing wood strength at the initial stage of brown rot through degradation of the primary cell wall.

scholarly journals Recent Advances in Screening Methods for the Functional Investigation of Lytic Polysaccharide Monooxygenases

2021 ◽  
Vol 9 ◽  
Author(s):  
Damao Wang ◽  
Yanping Li ◽  
Yuting Zheng ◽  
Yves S. Y. Hsieh
Keyword(s):  
Copper Ions ◽  
Biomass Conversion ◽  
Screening Methods ◽  
Lytic Polysaccharide Monooxygenase ◽  
Indirect Measurements ◽  
Glycosidic Bonds ◽  
Lytic Polysaccharide Monooxygenases ◽  
Polysaccharide Monooxygenases ◽  
Functional Investigation ◽  
Polysaccharide Monooxygenase

Lytic polysaccharide monooxygenase (LPMO) is a newly discovered and widely studied enzyme in recent years. These enzymes play a key role in the depolymerization of sugar-based biopolymers (including cellulose, hemicellulose, chitin and starch), and have a positive significance for biomass conversion. LPMO is a copper-dependent enzyme that can oxidize and cleave glycosidic bonds in cellulose and other polysaccharides. Their mechanism of action depends on the correct coordination of copper ions in the active site. There are still difficulties in the analysis of LPMO activity, which often requires multiple methods to be used in concert. In this review, we discussed various LPMO activity analysis methods reported so far, including mature mass spectrometry, chromatography, labeling, and indirect measurements, and summarized the advantages, disadvantages and applicability of different methods.

scholarly journals NEW ASPECTS OF THE ENZYMATIC BREAKDOWN OF CHITIN: A REVIEW

2021 ◽  
Vol 26 ◽  
pp. 5-10
Author(s):  
Urszula Jankiewicz ◽  
◽  
Arletta Kochańska-Jeziorska ◽  
Agnieszka Gałązka
Keyword(s):  
Crystalline Structure ◽  
Enzymatic Degradation ◽  
Soluble Sugars ◽  
Practical Importance ◽  
Oxidative Cleavage ◽  
Lytic Polysaccharide Monooxygenase ◽  
Great Practical Importance ◽  
Polysaccharide Monooxygenase

This review focuses on the enzymatic breakdown of chitin, taking into account the latest scientific reports on the activity of lytic polysaccharide monooxygenase (LPMO). Chitin is a natural, abundant polysaccharide of great practical importance in the environment. However, the insolubility in water and the tightly packed crystalline structure of chitin pose a serious obstacle to enzymatic degradation. This substrate can be converted into soluble sugars by the action of glycosidic hydrolases (GH), also known as chitinases. LPMO could prove to be helpful in enzymatic processes that increase the rate of chitin depolymerisation by improving the availability of substrates for chitinases. The unique action of LPMO is based on the ability to catalyse the oxidative cleavage of glycosidic chains present in complex, resistant crystal networks of chitin, and this cleavage facilitates the subsequent action of glycolytic hydrolases.

Production of functionalised chitins assisted by fungal lytic polysaccharide monooxygenase

2018 ◽  
Vol 20 (9) ◽  
pp. 2091-2100 ◽  
Author(s):  
Damao Wang ◽  
Jing Li ◽  
Germán Salazar-Alvarez ◽  
Lauren S. McKee ◽  
Vaibhav Srivastava ◽  
...  
Keyword(s):  
Fusarium Fujikuroi ◽  
Lytic Polysaccharide Monooxygenase ◽  
Polysaccharide Monooxygenase

The gene CCT67099 from Fusarium fujikuroi was shown to encode a novel enzyme from the Lytic Polysaccharide Monooxygenase (LPMO) Family AA11.

scholarly journals The H2O2-dependent activity of a fungal lytic polysaccharide monooxygenase investigated with a turbidimetric assay

2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Frantisek Filandr ◽  
Petr Man ◽  
Petr Halada ◽  
Hucheng Chang ◽  
Roland Ludwig ◽  
...  
Keyword(s):  
Lytic Polysaccharide Monooxygenase ◽  
Dependent Activity ◽  
Turbidimetric Assay ◽  
Polysaccharide Monooxygenase

Insecticidal fern protein Tma12 is possibly a lytic polysaccharide monooxygenase

Planta ◽  
2019 ◽  
Vol 249 (6) ◽  
pp. 1987-1996 ◽  
Author(s):  
Sunil K. Yadav ◽  
Archana ◽  
Rahul Singh ◽  
Pradhyumna Kumar Singh ◽  
Prema G. Vasudev
Keyword(s):  
Lytic Polysaccharide Monooxygenase ◽  
Polysaccharide Monooxygenase
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