The First Identification of Carbohydrate Binding Modules Specific to Chitosan

Shinya, Shoko; Ohnuma, Takayuki; Yamashiro, Reina; Kimoto, Hisashi; Kusaoke, Hideo; Anbazhagan, Padmanabhan; Juffer, André H.; Fukamizo, Tamo

doi:10.1074/jbc.m113.503243

Cited by 22 publications

(13 citation statements)

References 50 publications

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“…Substitution of Glu-38 by an aromatic amino acid Phe reduced the binding affinity drastically, suggesting the introduction of hydrophobic residues may alter the local conformation in the loop region affecting the chitosan-binding site. This finding was further supported by the observation that CBM32-2 of P. fukuinensis (16), which has a Tyr at this position, showed lower binding affinity toward chitosan oligomers.…”

Section: Discussionsupporting

confidence: 59%

“…Secondary structure for PeCBM32 is shown below the alignment. Binding Sites in PeCBM32-The affinity of CBM32 toward chitosan oligomers, (GlcN) 2 -(GlcN) 6 is reported using ITC (16). Here, we report the binding of GlcN with a stoichiometry of 2 and K b of 4 ϫ 10 3 M Ϫ1 , clearly indicating the accommodation of two GlcN residues by PeCBM32 (Table 3).…”

Section: Discussionmentioning

confidence: 54%

“…Specificity of PeCBM32-Many ligand molecules like galactose, lactose, sialic acid, LacNAc, chitosan oligomer, and mannan oligomer are recognized by CBM32 appended to different enzymes (7,16,26). We studied the influence of DA of chitosan polymers on the binding of PeCBM32.…”

Section: Discussionmentioning

confidence: 99%

“…To date, two tandemly occurring discoidin domains (DD1 and DD2) belonging to CBM32 from a chitosanase/glucanase of Paenibacillus fukuinensis are reported to have chitosan-binding ability (16). Binding experiments performed by ITC revealed that irrespective of high protein sequence identity (74%), DD1 and DD2 (hereafter denoted as CBM32-1 and CBM32-2) displayed different binding affinity and specificity toward chitosan oligosaccharides.…”

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confidence: 99%

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Amino Groups of Chitosan Are Crucial for Binding to a Family 32 Carbohydrate Binding Module of a Chitosanase from Paenibacillus elgii

Das¹,

Wagenknecht

Nareddy³

et al. 2016

Journal of Biological Chemistry

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We report here the role and mechanism of specificity of a family 32 carbohydrate binding module (CBM32) of a glycoside hydrolase family 8 chitosanase from Paenibacillus elgii (PeCsn). Both the activity and mode of action of PeCsn toward soluble chitosan polymers were not different with/without the CBM32 domain of P. elgii (PeCBM32). The decreased activity of PeCsn without PeCBM32 on chitosan powder suggested that PeCBM32 increases the relative concentration of enzyme on the substrate and thereby enhanced enzymatic activity. PeCBM32 specifically bound to polymeric and oligomeric chitosan and showed very weak binding to chitin and cellulose. In isothermal titration calorimetry, the binding stoichiometry of 2 and 1 for glucosamine monosaccharide (GlcN) and disaccharide (GlcN)2, respectively, was indicative of two binding sites in PeCBM32. A three-dimensional model-guided site-directed mutagenesis and the use of defined disaccharides varying in the pattern of acetylation suggested that the amino groups of chitosan and the polar residues Glu-16 and Glu-38 of PeCBM32 play a crucial role for the observed binding. The specificity of CBM32 has been further elucidated by a generated fusion protein PeCBM32-eGFP that binds to the chitosan exposing endophytic infection structures of Puccinia graminis f. sp. tritici Phylogenetic analysis showed that CBM32s appended to chitosanases are highly conserved across different chitosanase families suggesting their role in chitosan recognition and degradation. We have identified and characterized a chitosan-specific CBM32 useful for in situ staining of chitosans in the fungal cell wall during plant-fungus interaction.

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Section: Discussionsupporting

confidence: 59%

Section: Discussionmentioning

confidence: 54%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Amino Groups of Chitosan Are Crucial for Binding to a Family 32 Carbohydrate Binding Module of a Chitosanase from Paenibacillus elgii

Das¹,

Wagenknecht

Nareddy³

et al. 2016

Journal of Biological Chemistry

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“…However, new enzymes with exochitosanase activity have been reported, notably exo-β-D-glucosaminidase able to cleave CS from non-reducing termini, releasing GlcN residues [242,243]. Recently, the identification of a carbohydrate binding domain (CBM) for some chitosanases may suggest additional interaction with the CS polymer, involving a different mode of CS hydrolysis [244,245]. The chitosanases described are issued from many organisms, including bacteria, cyanobacteria, fungi, and plants [222].…”

Section: Biodegradability Of Chitosan Derivatives and Life Cycle Assementioning

confidence: 99%

Modification of Chitosan for the Generation of Functional Derivatives

et al. 2019

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Today, chitosan (CS) is probably considered as a biofunctional polysaccharide with the most notable growth and potential for applications in various fields. The progress in chitin chemistry and the need to replace additives and non-natural polymers with functional natural-based polymers have opened many new opportunities for CS and its derivatives. Thanks to the specific reactive groups of CS and easy chemical modifications, a wide range of physico-chemical and biological properties can be obtained from this ubiquitous polysaccharide that is composed of β-(1,4)-2-acetamido-2-deoxy-d-glucose repeating units. This review is presented to share insights into multiple native/modified CSs and chitooligosaccharides (COS) associated with their functional properties. An overview will be given on bioadhesive applications, antimicrobial activities, adsorption, and chelation in the wine industry, as well as developments in medical fields or biodegradability.

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Chitin/Chitosan-Active Enzymes Involved in Plant–Microbe Interactions

Fukamizo

Shinya

2019

Advances in Experimental Medicine and Biology

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The First Identification of Carbohydrate Binding Modules Specific to Chitosan

Cited by 22 publications

References 50 publications

Amino Groups of Chitosan Are Crucial for Binding to a Family 32 Carbohydrate Binding Module of a Chitosanase from Paenibacillus elgii

Amino Groups of Chitosan Are Crucial for Binding to a Family 32 Carbohydrate Binding Module of a Chitosanase from Paenibacillus elgii

Modification of Chitosan for the Generation of Functional Derivatives

Chitin/Chitosan-Active Enzymes Involved in Plant–Microbe Interactions

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