109957-89-3Relevant academic research and scientific papers
Depolymerization of β-chitin to mono- and disaccharides by the serum fraction from the para rubber tree, Hevea brasiliensis
Klaikherd, Akamol,Siripastr Jayanta,Boonjawat, Jariya,Aiba, Sei-Ichi,Sukwattanasinitt, Mongkol
, p. 2799 - 2804 (2004)
The serum fraction of latex from Hevea brasiliensis, the para rubber tree, is known to contain an endo-chitinolytic enzyme, hevamine. Herein the activity of the rubber serum towards β-chitin is investigated. The serum contained 6 mg/mL of protein and a chitinolytic activity of 18 mU per mg of protein. The optimum ratio of enzyme to chitin was 0.22 mU/mg, and the optimum substrate concentration was 60 mg/mL. The optimum pH range was pH 2-4, and the optimum temperature was 45°C. At these conditions both (GlcNAc)2 and GlcNAc were produced in a molar ratio of approximately 2:1. The hydrolysis of 300 mg of chitin with 64 mU of the rubber serum for 8 days under the optimum conditions gave 39 mg of GlcNAc and 108 mg of (GlcNAc)2 as determined by HPLC. Mixing the rubber serum preparation with an Aspergillus niger pectinase preparation containing β-N-acetylhexosaminidase can be used to produce almost exclusively the GlcNAc monomer in about 50% yield.
Enzymatic synthesis of 3-O-methylated chitin oligomers from new derivatives of a chitobiose oxazoline
Sakamoto, Junji,Kobayashi, Shiro
, p. 698 - 699 (2004)
Regiospecifically 3-O- and/or 3′-O-methylated derivatives of a chitobiose oxazoline have been synthesized as new substrate monomers and subjected to a chitinase catalysis, leading to the first synthesis of 3-O-methylated chitin oligomers via enzymatic oligomerization. (Graph Presented).
THE BEHAVIOR OF CHITIN TOWARDS ANHYDROUS HYDROGEN FLUORIDE. PREPARATION OF β(1-4)-LINKED 2-ACETAMIDO-2-DEOXY-D-GLUCOPYRANOSYL OLIGOSACCHARIDES
Bosso, Claude,Defaye, Jacques,Domard, Alain,Gadelle, Andree,Pedersen, Christian
, p. 57 - 68 (1986)
Fluorohydrolysis of chitin in anhydrous hydrogen fluoride led to β-(1-4)-linked 2-acetamido-2-deoxy-D-glucopyranosyl oligosaccharides in almost quantitative yield.The average d.p. depended on both reaction time and temperature, and was conveniently monito
Introduction of a tryptophan side chain into subsite +1 enhances transglycosylation activity of a GH-18 chitinase from Arabidopsis thaliana, AtChiC
Umemoto, Naoyuki,Ohnuma, Takayuki,Mizuhara, Mamiko,Sato, Hirokazu,Skriver, Karen,Fukamizo, Tamo
, p. 81 - 90 (2013)
A tryptophan side chain was introduced into subsite +1 of family GH-18 (class V) chitinases from Nicotiana tabacum and Arabidopsis thaliana (NtChiV and AtChiC, respectively) by the mutation of a glycine residue to tryptophan (G74W-NtChiV and G75W-AtChiC). The specific activity toward glycol chitin of the two mutant enzymes was 70-71% of that of the wild type. Using chitin oligosaccharides, (GlcNAc)n (n = 4, 5 and 6), as the substrates, we found the transglycosylation reaction to be significantly enhanced in G74W-NtChiV and G75W-AtChiC when compared with the corresponding wild-type enzymes. The introduced tryptophan side chain might protect the oxazolinium ion intermediate from attack by a nucleophilic water molecule. The enhancement of transglycosylation activity was much more distinct in G75W-AtChiC than in G74W-NtChiV. Nuclear magnetic resonance titration experiments using the inactive double mutants, E115Q/G74W-NtChiV and E116Q/G75W-AtChiC revealed that the association constant of (GlcNAc)5 was considerably larger for the latter. Amino acid substitutions at the acceptor binding site might have resulted in the larger association constant for G75W-AtChiC, giving rise to the higher transglycosylation activity of G75W-AtChiC. The Author 2012. Published by Oxford University Press. All rights reserved.
Enzymic synthesis of useful chito-oligosaccharides utilizing transglycosylation by chitinolytic enzymes in a buffer containing ammonium sulfate
Usui, Taichi,Matsui, Hidenori,Kiyoshi, Isobe
, p. 65 - 77 (1990)
A chitinase purified from culture filtrates of Trichoderma reesei KDR-11 efficiently catalyzed a transglycosylation reaction on tetra-N-acetylchitotetraoside in a buffer medium containing ammonium sulfate, converting the tetrasaccharide into hexa-N-acetylchitohexaose (39.6percent) and di-N-acetylchitobiose (55.7percent) as major products.Sugar-chain elongation from di-N-acetylchitobiose as the initial substrate to hexa-N-acetyl-chitohexaose and hepta-N-acetylchitoheptaose was also efficiently induced through lysozyme catalysis in the presence of ammonium sulfate at high (30percent) concentration.In this case, the addition of ammonium sulfate to the reaction system resulted in a remarkable increase of the hexamer and heptamer productions, which are desirable as biologically active oligosaccharides.
Expression and characterization of endochitinase C from Serratia marcescens BJL200 and its purification by a one-step general chitinase purification method
Synstad, Bjornar,Vaaje-Kolstad, Gustav,Cederkvist, F. Henning,Saua, Silje F.,Horn, Svein J.,Eijsink, Vincent G. H.,Sorlie, Morten
, p. 715 - 723 (2008)
In this study we cloned, expressed, purified, and charaterized chitinase C1 from Serratia marcescens strain BJL200. As expected, the BJL200-ChiC1 amino acid sequence of this strain was highly similar to sequences of ChiC1 identified in two other strains of S. marcescens. BJL200-ChiC1 was overproduced in E. coli by the T7 expression system, and purified by a one-step hydrophobic interaction chromatography (HIC) with phenyl-sepharose. BJL200-ChiA and BJL200-ChiB had an approximately 30-fold higher kcat and 15 fold-lower Km than BJL200-ChiC1 for the oligomeric substrate 4-methylumbelliferyl-β-D-N- N′-N″-triacetylchitotrioside, while BJL200-ChiC1 was 10-15 times faster than BJL200-ChiB and BJL200-ChiA in degrading the polymeric substrate CM-chitin-RBV. BJL200-ChiC1 degradation of β-chitin resulted in a range of different chito-oligosaccharides (GlcNAc)2 (main product), GlcNAc, (GlcNAc)3, (GlcNAc)4, and (GlcNAc)5, indicating endo activity. The purification method used for BJL200-ChiC1 in this study is generally applicable to family 18 chitinases and their mutants, including inactive mutants, some of which tend to bind almost irreversibly to chitin columns. The high specificity of the interaction with the (non-chitinous) column material is mediated by aromatic residues that occur in the substrate-binding clefts and surfaces of the enzymes.
Synthesis of derivatized chitooligomers using transglycosidases engineered from the fungal GH20 β-N-acetylhexosaminidase
Slámová, Kristyna,Krejzová, Jana,Marhol, Petr,Kalachova, Lubica,Kulik, Natallia,Pelantová, Helena,Cva?ka, Josef,K?en, Vladimír
, p. 1941 - 1950 (2015/06/02)
Abstract The synthesis of oligosaccharides using mutant glycosidases has been dynamically developing due to the need for novel carbohydrate-based materials. Chitooligomers (β-1→4-linked oligomers of N-acetylglucosamine) are bioactive compounds applicable in many industrial and pharmacological areas; however, their accessibility is still rather low. In this work, GH20 β-N-acetylhexosaminidase from the fungus Talaromyces flavus was engineered by site-directed mutagenesis to obtain three efficiently transglycosylating variants with ca. 200-times suppressed hydrolytic activity. Thus, we have prepared the first GH20 transglycosidases. In the reactions catalyzed by these mutant β-N-acetylhexosaminidases we were able to easily prepare and isolate both natural and modified chitooligomers in sufficient amounts for their complete spectral characterization and possible further application. The presented method for the synthesis of chitooligomers with aglycones suitable for linking to other biological structures is simple and robust enough to be easily scaled up.
Catalytic Depolymerization of Chitin with Retention of N-Acetyl Group
Yabushita, Mizuho,Kobayashi, Hirokazu,Kuroki, Kyoichi,Ito, Shogo,Fukuoka, Atsushi
, p. 3760 - 3763 (2015/12/08)
Chitin, a polymer of N-acetylglucosamine units with β-1,4-glycosidic linkages, is the most abundant marine biomass. Chitin monomers containing N-acetyl groups are useful precursors to various fine chemicals and medicines. However, the selective conversion of robust chitin to N-acetylated monomers currently requires a large excess of acid or a long reaction time, which limits its application. We demonstrate a fast catalytic transformation of chitin to monomers with retention of N-acetyl groups by combining mechanochemistry and homogeneous catalysis. Mechanical-force-assisted depolymerization of chitin with a catalytic amount of H2SO4 gave soluble short-chain oligomers. Subsequent hydrolysis of the ball-milled sample provided N-acetylglucosamine in 53 % yield, and methanolysis afforded 1-O-methyl-N-acetylglucosamine in yields of up to 70 %. Our process can greatly reduce the use of acid compared to the conventional process.
An ammonium sulfate sensitive chitinase from Streptomyces sp. CS501
Rahman, Md. Arifur,Choi, Yun Hee,Pradeep,Yoo, Jin Cheol
, p. 1522 - 1529 (2015/03/30)
A chitinase from Streptomyces sp. CS501 was isolated from the Korean soil sample, purified by single-step chromatography, and biochemically characterized. The extracellular chitinase (Ch501) was purified to 4.60 fold with yield of 28.74 % using Sepharose Cl-6B column. The molecular mass of Ch501 was approximately 43 kDa as estimated by SDS-PAGE and zymography. The enzyme (Ch501) was found to be stable over a broad pH range (5.0-10.0) and temperature (up to 50 °C), and have an optimum temperature of 60 °C. N-terminal sequence of Ch501 was AAYDDAAAAA. Intriguingly, Ch501 was highly sensitive to ammonium sulfate but it's completely suppressed activity was recovered after desalting out. TLC analysis of Ch501 showed the production of N-acetyl d-glucosamine (GlcNAc) and Diacetylchitobiose (GlcNAc)2, as a principal hydrolyzed product. Ch501 shows antifungal activity against Fusarium solani and Aspergillus brasiliensis, which can be used for the biological control of fungus. As has been simple in purification, stable in a broad range of pH, ability to produce oligosaccharides, and antifungal activity showed that Ch501 has potential applications in industries as for chitooligosaccharides production used as prebiotics and/or for the biological control of plant pathogens in agriculture.
Development of innovative technologies to decrease the environmental burdens associated with using chitin as a biomass resource: Mechanochemical grinding and enzymatic degradation
Nakagawa, Yuko S.,Oyama, Yasuhiro,Kon, Nobuko,Nikaido, Mitsuru,Tanno, Koichi,Kogawa, Jun,Inomata, Shoji,Masui, Ayano,Yamamura, Akihiro,Kawaguchi, Mitsuaki,Matahira, Yoshiharu,Totani, Kazuhide
experimental part, p. 1843 - 1849 (2011/09/14)
The production of N-acetylglucosamine (GlcNAc, chitin monosaccharide) from crab or shrimp shells generally requires numerous steps and the use of deleterious substances. One goal for researchers is the direct production of GlcNAc and NN′-diacetylchitobiose [(GlcNAc)2, chitin structural dimeric unit] from both chitin and crab shells. The present study reports the development of an intensive ball "converge" mill for the rapid mechanochemical conversion of chitin or crab shells into amorphous, chitinase-sensitive microparticles. Optimal crab shell grinding parameters were determined, and close to 100% direct degradation of chitin from crab shell to GlcNAc was achieved. This is the first report of using a mechanochemical process with enzymatic degradation to decrease the environmental burdens associated with GlcNAc production from chitin.
