EzCatDB: S00265

DB codeS00265
RLCP classification1.30.300.2
CATH domainDomain 11.10.530.40Catalytic domain
E.C.3.2.1.132
CSA1chk

CATH domainRelated DB codes (homologues)
1.10.530.40S00021

Enzyme Name
Swiss-protKEGG

P33673P33665
Protein nameChitosanaseChitosanasechitosanase
SynonymsEC 3.2.1.132
EC 3.2.1.132

KEGG pathways
MAP codePathways
MAP00530Aminosugars metabolism

Swiss-prot:Accession NumberP33673P33665
Entry nameCHIS_BACCICHIS_STRSN
ActivityEndohydrolysis of beta-(1->4)-linkages between D-glucosamine residues in a partly acetylated chitosan.Endohydrolysis of beta-(1->4)-linkages between D-glucosamine residues in a partly acetylated chitosan.
Subunit

Subcellular locationSecreted.Secreted.
Cofactor



SubstratesProducts
KEGG-idC00734C00001C00734C06023
CompoundChitosanH2OChitosanD-Glucosaminide
Typeamine group,polysaccharideH2Oamine group,polysaccharideamine group,polysaccharide
1chkAUnbound
UnboundUnbound
1chkBUnbound
UnboundUnbound
1qgiAAnalogue:GCS-GCS-NAG
UnboundUnbound

Active-site residues
resource
Swiss-prot;P33665, P33673 & literature [4], [6]
pdbCatalytic residues
1chkAGLU 22;ASP 40
1chkBGLU 22;ASP 40
1qgiAGLU 37;ASP 55

References for Catalytic Mechanism
ReferencesSectionsNo. of steps in catalysis
[4]Fig.7, p.158-160
[6]Fig.2, p.690-691

references
[1]
CommentsCHARACTERIZATION.
Medline ID96033029
PubMed ID7487871
JournalBiochem J
Year1995
Volume311
Pages377-83
AuthorsFukamizo T, Honda Y, Goto S, Boucher I, Brzezinski R
TitleReaction mechanism of chitosanase from Streptomyces sp. N174.
Related Swiss-protP33665
[2]
CommentsMUTAGENESIS.
Medline ID96125086
PubMed ID8537367
JournalJ Biol Chem
Year1995
Volume270
Pages31077-82
AuthorsBoucher I, Fukamizo T, Honda Y, Willick GE, Neugebauer WA, Brzezinski R
TitleSite-directed mutagenesis of evolutionary conserved carboxylic amino acids in the chitosanase from Streptomyces sp. N174 reveals two residues essential for catalysis.
Related Swiss-protP33665
[3]
PubMed ID8535779
JournalStructure
Year1995
Volume3
Pages853-9
AuthorsDavies G, Henrissat B
TitleStructures and mechanisms of glycosyl hydrolases.
[4]
CommentsX-RAY CRYSTALLOGRAPHY (2.4 ANGSTROMS).
Medline ID96163435
PubMed ID8564542
JournalNat Struct Biol
Year1996
Volume3
Pages155-62
AuthorsMarcotte EM, Monzingo AF, Ernst SR, Brzezinski R, Robertus JD
TitleX-ray structure of an anti-fungal chitosanase from streptomyces N174.
Related PDB1chk
Related Swiss-protP33665
[5]
PubMed ID8564539
JournalNat Struct Biol
Year1996
Volume3
Pages133-40
AuthorsMonzingo AF, Marcotte EM, Hart PJ, Robertus JD
TitleChitinases, chitosanases, and lysozymes can be divided into procaryotic and eucaryotic families sharing a conserved core.
[6]
PubMed ID9599657
JournalBiochem Cell Biol
Year1997
Volume75
Pages687-96
AuthorsFukamizo T, Brzezinski R
TitleChitosanase from Streptomyces sp. strain N174: a comparative review of its structure and function.
[7]
PubMed ID9989221
JournalBiochim Biophys Acta
Year1999
Volume1429
Pages365-76
AuthorsHonda Y, Fukamizo T, Okajima T, Goto S, Boucher I, Brzezinski R
TitleThermal unfolding of chitosanase from Streptomyces sp. N174: role of tryptophan residues in the protein structure stabilization.
[8]
CommentsSTRAIN=MH-K1;
Medline ID99452978
PubMed ID10521473
JournalJ Biol Chem
Year1999
Volume274
Pages30818-25
AuthorsSaito J, Kita A, Higuchi Y, Nagata Y, Ando A, Miki K
TitleCrystal structure of chitosanase from Bacillus circulans MH-K1 at 1.6-A resolution and its substrate recognition mechanism.
Related PDB1qgi
Related Swiss-protP33673
[9]
PubMed ID12369923
JournalCurr Protein Pept Sci
Year2000
Volume1
Pages105-24
AuthorsFukamizo T
TitleChitinolytic enzymes: catalysis, substrate binding, and their application.
[10]
PubMed ID10829022
JournalJ Biol Chem
Year2000
Volume275
Pages25633-40
AuthorsFukamizo T, Juffer AH, Vogel HJ, Honda Y, Tremblay H, Boucher I, Neugebauer WA, Brzezinski R
TitleTheoretical calculation of pKa reveals an important role of Arg205 in the activity and stability of Streptomyces sp. N174 chitosanase.
[11]
PubMed ID11499927
JournalAppl Microbiol Biotechnol
Year2001
Volume56
Pages173-80
AuthorsYoon HG, Kim HY, Lim YH, Kim HK, Shin DH, Hong BS, Cho HY
TitleIdentification of essential amino acid residues for catalytic activity and thermostability of novel chitosanase by site-directed mutagenesis.
[12]
PubMed ID12092850
JournalBiosci Biotechnol Biochem
Year2002
Volume66
Pages986-95
AuthorsYoon HG, Lee KH, Kim HY, Kim HK, Shin DH, Hong BS, Cho HY
TitleGene cloning and biochemical analysis of thermostable chitosanase (TCH-2) from Bacillus coagulans CK108.
[13]
PubMed ID11754739
JournalJ Biochem (Tokyo)
Year2002
Volume131
Pages87-96
AuthorsShimono K, Shigeru K, Tsuchiya A, Itou N, Ohta Y, Tanaka K, Nakagawa T, Matsuda H, Kawamukai M
TitleTwo glutamic acids in chitosanase A from Matsuebacter chitosanotabidus 3001 are the catalytically important residues.

comments
This enzyme belongs to the glycosidase family-46, with an inverting mechanism.
According to the literature [4] & [6], the catalytic reaction proceeds by an SN2 mechanism, as follows:
(1) Asp40 (of PDB;1chk) acts as a general base, which activates a water molecule to attack on the C1 atom of the glycoside bond to be cleaved. Here, the sugar can distort to a half-chair configuration for the nucleophilic attack.
(2) At the same time, Glu22 acts as a general acid to protonate the leaving group, which facilitate the nucleophilic attack by the water.

createdupdated
2005-03-232009-02-26
Copyright: Nozomi Nagano, JST & CBRC-AIST
Funded by PRESTO/Japan Science and Technology Corporation (JST) (December 2001 - November 2004)
Funded by Grant-in-Aid for Publication of Scientific Research Results/Japan Society for the Promotion of Science (JSPS) (April 2005 - March 2006)
Funded by Grant-in-Aid for Scientific Research (B)/Japan Society for the Promotion of Science (JSPS) (April 2005 - March 2008)
Funded by BIRD/Japan Science and Technology Corporation (JST) (September 2005 - September 2010)
Funded by BIRD/Japan Science and Technology Corporation (JST) (October 2007 - September 2010)
Funded by Grant-in-Aid for Publication of Scientific Research Results/Japan Society for the Promotion of Science (JSPS) (April 2011 - March 2012)

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