For Publishers
DiscoveryMetadataPeer reviewHostingPublishing
For Researchers
JoinPublishReviewCollect
Register
Blog
About
Search
Advanced search
My ScienceOpen
Search
For Publishers
For Researchers
Blog
About
6
views
32
references
 Top references
cited by
8
    
0 reviews
 Review
0
comments
 Comment
0
recommends
+1 Recommend
0 collections
    0
    shares
      140
      similar
       All similar
      • Record: found
      • Abstract: found
      • Article: found
      Is Open Access

      Role of N-linked glycosylation in the enzymatic properties of a thermophilic GH 10 xylanase from Aspergillus fumigatus expressed in Pichia pastoris

      research-article

      Read this article at

      ScienceOpenPublisherPMC
      Bookmark
          There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.

          Abstract

          N-Glycosylation is a posttranslational modification commonly occurred in fungi and plays roles in a variety of enzyme functions. In this study, a xylanase ( Af-XYNA) of glycoside hydrolase (GH) family 10 from Aspergillus fumigatus harboring three potential N-glycosylation sites (N87, N124 and N335) was heterologously produced in Pichia pastoris. The N-glycosylated Af-XYNA (WT) exhibited favorable temperature and pH optima (75°C and pH 5.0) and good thermostability (maintaining stable at 60°C). To reveal the role of N-glycosylation on Af-XYNA, the enzyme was deglycosylated by endo-β-N-acetylglucosaminidase H (DE) or modified by site-directed mutagenesis at N124 (N124T). The deglycosylated DE and mutant N124T showed narrower pH adaptation range, lower specific activity, and worse pH and thermal stability. Further thermodynamic analysis revealed that the enzyme with higher N-glycosylation degree was more thermostable. This study demonstrated that the effects of glycosylation at different degrees and sites were diverse, in which the glycan linked to N124 played a key role in pH and thermal stability of Af-XYNA.

          Related collections

          Most cited references32

          • Record: found
          • Abstract: found
          • Article: not found

          Xylanases from fungi: properties and industrial applications.

          M L T M Polizeli, A C S Rizzatti, R Monti … (2005)
          Xylan is the principal type of hemicellulose. It is a linear polymer of beta-D-xylopyranosyl units linked by (1-4) glycosidic bonds. In nature, the polysaccharide backbone may be added to 4-O-methyl-alpha-D-glucuronopyranosyl units, acetyl groups, alpha-L-arabinofuranosyl, etc., in variable proportions. An enzymatic complex is responsible for the hydrolysis of xylan, but the main enzymes involved are endo-1,4-beta-xylanase and beta-xylosidase. These enzymes are produced by fungi, bacteria, yeast, marine algae, protozoans, snails, crustaceans, insect, seeds, etc., but the principal commercial source is filamentous fungi. Recently, there has been much industrial interest in xylan and its hydrolytic enzymatic complex, as a supplement in animal feed, for the manufacture of bread, food and drinks, textiles, bleaching of cellulose pulp, ethanol and xylitol production. This review describes some properties of xylan and its metabolism, as well as the biochemical properties of xylanases and their commercial applications.
          Article 0 comments Cited 211 times – based on 0 reviews     Review now
            Bookmark
            • Record: found
            • Abstract: found
            • Article: not found

            Microbial xylanases: engineering, production and industrial applications.

            Veeresh Juturu, Jin Chuan Wu (2012)
            Enzymatic depolymerization of hemicellulose to monomer sugars needs the synergistic action of multiple enzymes, among them endo-xylanases (EC 3.2.1.8) and β-xylosidases (EC 3.2.1.37) (collectively xylanases) play a vital role in depolymerizing xylan, the major component of hemicellulose. Recent developments in recombinant protein engineering have paved the way for engineering and expressing xylanases in both heterologous and homologous hosts. Functional expression of endo-xylanases has been successful in many hosts including bacteria, yeasts, fungi and plants with yeasts being the most promising expression systems. Functional expression of β-xylosidases is more challenging possibly due to their more complicated structures. The structures of endo-xylanases of glycoside hydrolase families 10 and 11 have been well elucidated. Family F/10 endo-xylanases are composed of a cellulose-binding domain and a catalytic domain connected by a linker peptide with a (β/α)8 fold TIM barrel. Family G/11 endo-xylanases have a β-jelly roll structure and are thought to be able to pass through the pores of hemicellulose network owing to their smaller molecular sizes. The structure of a β-D-xylosidase belonging to family 39 glycoside hydrolase has been elucidated as a tetramer with each monomer being composed of three distinct regions: a catalytic domain of the canonical (β/α)8--TIM barrel fold, a β-sandwich domain and a small α-helical domain with the enzyme active site that binds to D-xylooligomers being present on the upper side of the barrel. Glycosylation is generally considered as one of the most important post-translational modifications of xylanases, but a few examples showed functional expression of eukaryotic xylanases in bacteria. The optimal ratio of these synergistic enzymes is very important in improving hydrolysis efficiency and reducing enzyme dosage but has hardly been addressed in literature. Xylanases have been used in traditional fields such as food, feed and paper industries for a longer time but more and more attention has been paid to using them in producing sugars and other chemicals from lignocelluloses in recent years. Mining new genes from nature, rational engineering of known genes and directed evolution of these genes are required to get tailor-made xylanases for various industrial applications. Copyright © 2011 Elsevier Inc. All rights reserved.
            Article 0 comments Cited 77 times – based on 0 reviews     Review now
              Bookmark
              • Record: found
              • Abstract: found
              • Article: not found

              Thermostable enzymes in lignocellulose hydrolysis.

              Liisa Viikari, Marika Alapuranen, Terhi Puranen … (2007)
              Thermostable enzymes offer potential benefits in the hydrolysis of lignocellulosic substrates; higher specific activity decreasing the amount of enzymes, enhanced stability allowing improved hydrolysis performance and increased flexibility with respect to process configurations, all leading to improvement of the overall economy of the process. New thermostable cellulase mixtures were composed of cloned fungal enzymes for hydrolysis experiments. Three thermostable cellulases, identified as the most promising enzymes in their categories (cellobiohydrolase, endoglucanase and beta-glucosidase), were cloned and produced in Trichoderma reesei and mixed to compose a novel mixture of thermostable cellulases. Thermostable xylanase was added to enzyme preparations used on substrates containing residual hemicellulose. The new optimised thermostable enzyme mixtures were evaluated in high temperature hydrolysis experiments on technical steam pretreated raw materials: spruce and corn stover. The hydrolysis temperature could be increased by about 10-15 degrees C, as compared with present commercial Trichoderma enzymes. The same degree of hydrolysis, about 90% of theoretical, measured as individual sugars, could be obtained with the thermostable enzymes at 60 degrees C as with the commercial enzymes at 45 degrees C. Clearly more efficient hydrolysis per assayed FPU unit or per amount of cellobiohydrolase I protein used was obtained. The maximum FPU activity of the novel enzyme mixture was about 25% higher at the optimum temperature at 65 degrees C, as compared with the highest activity of the commercial reference enzyme at 60 degrees C. The results provide a promising basis to produce and formulate improved enzyme products. These products can have high temperature stability in process conditions in the range of 55-60 degrees C (with present industrial products at 45-50 degrees C) and clearly improved specific activity, essentially decreasing the protein dosage required for an efficient hydrolysis of lignocellulosic substrates. New types of process configurations based on thermostable enzymes are discussed.
              Article 0 comments Cited 67 times – based on 0 reviews     Review now
                Bookmark
                All references

                Author and article information

                Contributors
                Olaf Kniemeyer: Role: Editor
                Journal
                Journal ID (nlm-ta): PLoS One
                Journal ID (iso-abbrev): PLoS ONE
                Journal ID (publisher-id): plos
                Journal ID (pmc): plosone
                Title: PLoS ONE
                Publisher: Public Library of Science (San Francisco, CA USA )
                ISSN (Electronic): 1932-6203
                Publication date (Electronic): 10 February 2017
                Publication date Collection: 2017
                Volume: 12
                Issue: 2
                Electronic Location Identifier: e0171111
                Affiliations
                [1 ]College of Biological Sciences and Engineering, Jiangxi Agricultural University, Nanchang, People’s Republic of China
                [2 ]Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing, People’s Republic of China
                Leibniz-Institut fur Naturstoff-Forschung und Infektionsbiologie eV Hans-Knoll-Institut, GERMANY
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                • Conceptualization: PS BY.

                • Data curation: PS RM.

                • Formal analysis: XC PS.

                • Funding acquisition: BX PS BY.

                • Investigation: PS BY.

                • Methodology: XC HL.

                • Project administration: PS.

                • Resources: XC PS BX.

                • Software: HL YB.

                • Supervision: PS YB.

                • Validation: BY.

                • Visualization: PS.

                • Writing – original draft: XC PS BX.

                • Writing – review & editing: RM BY.

                Article
                Publisher ID: PONE-D-16-27678
                DOI: 10.1371/journal.pone.0171111
                PMC ID: 5302446
                PubMed ID: 28187141
                SO-VID: b5370d50-bbff-4049-8039-ae0ff8c46365
                Copyright © © 2017 Chang et al
                License:

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                Date received : 15 July 2016
                Date accepted : 15 January 2017
                Page count
                Figures: 4, Tables: 1, Pages: 13
                Funding
                Funded by: the National Natural Science Foundation of China
                Award ID: 31372345
                Award Recipient :
                Funded by: the National Natural Science Foundation of China
                Award ID: 31360372
                Award Recipient :
                Funded by: the National Science Fund for Distinguished Young Scholars
                Award ID: 31225026
                Award Recipient :
                Funded by: the China Modern Agriculture Research System
                Award ID: CARS-42
                Award Recipient :
                Financial support includes the National Natural Science Foundation of China (grant numbers 31372345 (PS) and 31360372 (BX); [ http://www.nsfc.gov.cn/]), the National Science Fund for Distinguished Young Scholars (grant number 31225026 (BY); [ http://www.nsfc.gov.cn/]), and the China Modern Agriculture Research System (grant number CARS-42 (BY); [ http://www.zgrj.org/]). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Subject: Research Article
                Subject: Biology and Life Sciences
                Subject: Biochemistry
                Subject: Glycobiology
                Subject: Glycosylation
                Subject: Biology and Life Sciences
                Subject: Biochemistry
                Subject: Proteins
                Subject: Post-Translational Modification
                Subject: Glycosylation
                Subject: Biology and Life Sciences
                Subject: Agriculture
                Subject: Crop Science
                Subject: Crops
                Subject: Cereal Crops
                Subject: Wheat
                Subject: Biology and Life Sciences
                Subject: Organisms
                Subject: Plants
                Subject: Grasses
                Subject: Wheat
                Subject: Computer and Information Sciences
                Subject: Computer Software
                Subject: Biology and Life Sciences
                Subject: Organisms
                Subject: Fungi
                Subject: Yeast
                Subject: Pichia Pastoris
                Subject: Biology and Life Sciences
                Subject: Organisms
                Subject: Fungi
                Subject: Physical Sciences
                Subject: Chemistry
                Subject: Chemical Reactions
                Subject: Hydrolysis
                Subject: Biology and Life Sciences
                Subject: Biochemistry
                Subject: Proteins
                Subject: Post-Translational Modification
                Subject: Signal Peptides
                Subject: Biology and Life Sciences
                Subject: Organisms
                Subject: Fungi
                Subject: Molds (Fungi)
                Subject: Aspergillus
                Subject: Aspergillus Fumigatus
                Subject: Biology and Life Sciences
                Subject: Microbiology
                Subject: Medical Microbiology
                Subject: Microbial Pathogens
                Subject: Fungal Pathogens
                Subject: Aspergillus Fumigatus
                Subject: Medicine and Health Sciences
                Subject: Pathology and Laboratory Medicine
                Subject: Pathogens
                Subject: Microbial Pathogens
                Subject: Fungal Pathogens
                Subject: Aspergillus Fumigatus
                Subject: Biology and Life Sciences
                Subject: Mycology
                Subject: Fungal Pathogens
                Subject: Aspergillus Fumigatus
                Custom metadata
                Data Availability All relevant data are within the paper.

                ScienceOpen disciplines: Uncategorized
                Data availability:
                ScienceOpen disciplines: Uncategorized

                Comments

                Comment on this article

                scite_

                Similar content140

                See all similar

                Cited by8

                See all cited by

                Most referenced authors640

                See all reference authors