GUN_CRYAT
ID GUN_CRYAT Reviewed; 225 AA.
AC D3GDK4;
DT 12-AUG-2020, integrated into UniProtKB/Swiss-Prot.
DT 23-MAR-2010, sequence version 1.
DT 25-MAY-2022, entry version 43.
DE RecName: Full=Endoglucanase {ECO:0000305};
DE EC=3.2.1.4 {ECO:0000255|PROSITE-ProRule:PRU10069, ECO:0000269|PubMed:22333528, ECO:0000269|PubMed:24848382, ECO:0000269|PubMed:28156112};
DE AltName: Full=CaCel {ECO:0000303|PubMed:22333528, ECO:0000303|PubMed:24848382, ECO:0000303|PubMed:28156112};
DE AltName: Full=Cellulase {ECO:0000255|PROSITE-ProRule:PRU10069, ECO:0000303|PubMed:22333528, ECO:0000303|PubMed:24848382, ECO:0000303|PubMed:28156112};
DE AltName: Full=Endo-beta-1,4-glucanase {ECO:0000303|PubMed:24848382, ECO:0000303|PubMed:28156112, ECO:0000312|EMBL:ACV50414.1};
DE Flags: Precursor;
OS Cryptopygus antarcticus (Antarctic springtail).
OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Collembola;
OC Entomobryomorpha; Isotomoidea; Isotomidae; Anurophorinae; Cryptopygus;
OC Cryptopygus antarcticus complex.
OX NCBI_TaxID=187623 {ECO:0000312|EMBL:ACV50414.1};
RN [1] {ECO:0000312|EMBL:ACV50414.1, ECO:0000312|EMBL:ACV50415.1}
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA / MRNA].
RA Song J.M., Lee Y.-H.;
RT "Molecular cloning and characterization of a novel endo-beta-1,4-glucanase
RT from the Antarctic springtail, Cryptopygus antarcticus.";
RL Submitted (JAN-2009) to the EMBL/GenBank/DDBJ databases.
RN [2]
RP PROTEIN SEQUENCE OF 16-25, FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL
RP PROPERTIES, SUBCELLULAR LOCATION, GLYCOSYLATION, AND BIOTECHNOLOGY.
RX PubMed=24848382; DOI=10.1007/s12033-014-9767-8;
RA Hong S.M., Sung H.S., Kang M.H., Kim C.G., Lee Y.H., Kim D.J., Lee J.M.,
RA Kusakabe T.;
RT "Characterization of Cryptopygus antarcticus endo-beta-1,4-glucanase from
RT Bombyx mori expression systems.";
RL Mol. Biotechnol. 56:878-889(2014).
RN [3]
RP CATALYTIC ACTIVITY.
RX PubMed=22333528; DOI=10.1016/j.pep.2012.01.020;
RA Song J.M., An Y.J., Kang M.H., Lee Y.H., Cha S.S.;
RT "Cultivation at 6-10 degrees C is an effective strategy to overcome the
RT insolubility of recombinant proteins in Escherichia coli.";
RL Protein Expr. Purif. 82:297-301(2012).
RN [4] {ECO:0007744|PDB:5H4U}
RP X-RAY CRYSTALLOGRAPHY (2.60 ANGSTROMS), FUNCTION, CATALYTIC ACTIVITY,
RP ACTIVITY REGULATION, BIOPHYSICOCHEMICAL PROPERTIES, SUBSTRATE SPECIFICITY,
RP BIOTECHNOLOGY, AND DISULFIDE BONDS.
RX PubMed=28156112; DOI=10.1021/acs.jafc.6b05037;
RA Song J.M., Hong S.K., An Y.J., Kang M.H., Hong K.H., Lee Y.H., Cha S.S.;
RT "Genetic and Structural Characterization of a Thermo-Tolerant, Cold-Active,
RT and Acidic Endo-beta-1,4-glucanase from Antarctic Springtail, Cryptopygus
RT antarcticus.";
RL J. Agric. Food Chem. 65:1630-1640(2017).
CC -!- FUNCTION: Hydrolyzes carboxymethylcellulose (CMC) (PubMed:24848382,
CC PubMed:28156112). Hydrolyzes also lichenan and barley beta-1,4-D-
CC glucan. CMC is hydrolyzed majorily to cellobiose (G2), cellotriose (G3)
CC and cellotetraose (G4). Cellohexaose (G6) is hydrolyzed to G4 and G2
CC with traces of G3. Cellopentaose (G5) is completely hydrolyzed to G2
CC and G3, and G4 is partially hydrolyzed to G2. Does not hydrolyze G2 or
CC G3. Does not hydrolyze crystalline cellulose, soluble starch, xylan,
CC mannan or laminarin (PubMed:28156112). {ECO:0000269|PubMed:24848382,
CC ECO:0000269|PubMed:28156112}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Endohydrolysis of (1->4)-beta-D-glucosidic linkages in
CC cellulose, lichenin and cereal beta-D-glucans.; EC=3.2.1.4;
CC Evidence={ECO:0000255|PROSITE-ProRule:PRU10069,
CC ECO:0000269|PubMed:22333528, ECO:0000269|PubMed:24848382,
CC ECO:0000269|PubMed:28156112};
CC -!- ACTIVITY REGULATION: Activity is not affected by metal ions except
CC Mn(2+), which reduces the activity by 40-50%. However, no significant
CC change in activity in response to 1 mM EDTA.
CC {ECO:0000269|PubMed:28156112}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC pH dependence:
CC Optimum pH is 3.5 (PubMed:24848382, PubMed:28156112). More than 90%
CC of the maximal activity is maintained between pH range 3-5
CC (PubMed:24848382). More than 60% of the maximal activity is
CC maintained between pH range 2-8 (PubMed:24848382, PubMed:28156112).
CC About 80% of the maximal activity is maintained even at pH 2.5
CC (PubMed:28156112). {ECO:0000269|PubMed:24848382,
CC ECO:0000269|PubMed:28156112};
CC Temperature dependence:
CC Optimum temperature is around 40-50 degrees Celsius (PubMed:24848382,
CC PubMed:28156112). Has more than 50% of the maximal activity between
CC 10-70 degrees Celsius, and more than 30% of activity at 0 degrees
CC Celsius. Stable at 50 degrees Celsius for 30 min (PubMed:24848382).
CC 60-80% of the maximal activity is retained between 0-10 degrees
CC Celsius. Displays 40% of its maximal activity at as high as 80
CC degrees Celsius. Remains active at 60 degrees Celsius for over 8
CC hours. Approximately 50% of the activity is still maintained after 4-
CC hour incubation at 70 degrees Celsius (PubMed:28156112).
CC {ECO:0000269|PubMed:24848382, ECO:0000269|PubMed:28156112};
CC -!- SUBCELLULAR LOCATION: Secreted {ECO:0000269|PubMed:24848382}.
CC -!- PTM: N- and O-glycosylated. Contains hybrid- and complex-type N-
CC glycans. {ECO:0000269|PubMed:24848382}.
CC -!- BIOTECHNOLOGY: Potential use as a cellulase in industry due to its high
CC catalytic activity at low temperature as well as its thermotolerance
CC (PubMed:24848382). Digests green algae (Ulva pertusa) under acidic
CC conditions at 50 degrees Celsius, which suggests that it could be used
CC for biofuel production from seaweeds (PubMed:28156112).
CC {ECO:0000305|PubMed:24848382, ECO:0000305|PubMed:28156112}.
CC -!- SIMILARITY: Belongs to the glycosyl hydrolase 45 (cellulase K) family.
CC {ECO:0000305}.
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DR EMBL; FJ648735; ACV50414.1; -; mRNA.
DR EMBL; FJ648736; ACV50415.1; -; Genomic_DNA.
DR PDB; 5H4U; X-ray; 2.60 A; A/B/C=1-225.
DR PDBsum; 5H4U; -.
DR AlphaFoldDB; D3GDK4; -.
DR SMR; D3GDK4; -.
DR CAZy; GH45; Glycoside Hydrolase Family 45.
DR BRENDA; 3.2.1.4; 11159.
DR GO; GO:0005576; C:extracellular region; IDA:UniProtKB.
DR GO; GO:0008810; F:cellulase activity; IDA:UniProtKB.
DR GO; GO:0030245; P:cellulose catabolic process; IDA:UniProtKB.
DR Gene3D; 2.40.40.10; -; 1.
DR InterPro; IPR000334; Glyco_hydro_45.
DR InterPro; IPR036908; RlpA-like_sf.
DR Pfam; PF02015; Glyco_hydro_45; 1.
DR SUPFAM; SSF50685; SSF50685; 1.
DR PROSITE; PS01140; GLYCOSYL_HYDROL_F45; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Carbohydrate metabolism; Cellulose degradation;
KW Direct protein sequencing; Disulfide bond; Glycoprotein; Glycosidase;
KW Hydrolase; Polysaccharide degradation; Secreted; Signal.
FT SIGNAL 1..15
FT /evidence="ECO:0000269|PubMed:24848382"
FT CHAIN 16..225
FT /note="Endoglucanase"
FT /evidence="ECO:0000305|PubMed:24848382"
FT /id="PRO_5010494529"
FT ACT_SITE 29
FT /note="Nucleophile"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU10069,
FT ECO:0000305|PubMed:28156112"
FT ACT_SITE 138
FT /note="Proton donor"
FT /evidence="ECO:0000305|PubMed:28156112"
FT CARBOHYD 55
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00498"
FT DISULFID 30..152
FT /evidence="ECO:0000269|PubMed:28156112,
FT ECO:0007744|PDB:5H4U"
FT DISULFID 31..66
FT /evidence="ECO:0000269|PubMed:28156112,
FT ECO:0007744|PDB:5H4U"
FT DISULFID 35..103
FT /evidence="ECO:0000269|PubMed:28156112,
FT ECO:0007744|PDB:5H4U"
FT DISULFID 50..74
FT /evidence="ECO:0000269|PubMed:28156112,
FT ECO:0007744|PDB:5H4U"
FT DISULFID 104..219
FT /evidence="ECO:0000269|PubMed:28156112,
FT ECO:0007744|PDB:5H4U"
FT DISULFID 106..209
FT /evidence="ECO:0000269|PubMed:28156112,
FT ECO:0007744|PDB:5H4U"
FT DISULFID 176..187
FT /evidence="ECO:0000269|PubMed:28156112,
FT ECO:0007744|PDB:5H4U"
FT CONFLICT 16..17
FT /note="GL -> MV (in Ref. 2; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 16
FT /note="G -> I (in Ref. 2; AA sequence)"
FT /evidence="ECO:0000305"
FT STRAND 19..27
FT /evidence="ECO:0007829|PDB:5H4U"
FT HELIX 34..36
FT /evidence="ECO:0007829|PDB:5H4U"
FT STRAND 40..45
FT /evidence="ECO:0007829|PDB:5H4U"
FT STRAND 54..57
FT /evidence="ECO:0007829|PDB:5H4U"
FT HELIX 65..67
FT /evidence="ECO:0007829|PDB:5H4U"
FT STRAND 69..72
FT /evidence="ECO:0007829|PDB:5H4U"
FT HELIX 75..77
FT /evidence="ECO:0007829|PDB:5H4U"
FT STRAND 80..82
FT /evidence="ECO:0007829|PDB:5H4U"
FT STRAND 85..96
FT /evidence="ECO:0007829|PDB:5H4U"
FT HELIX 99..102
FT /evidence="ECO:0007829|PDB:5H4U"
FT STRAND 106..111
FT /evidence="ECO:0007829|PDB:5H4U"
FT TURN 115..118
FT /evidence="ECO:0007829|PDB:5H4U"
FT STRAND 120..128
FT /evidence="ECO:0007829|PDB:5H4U"
FT STRAND 133..140
FT /evidence="ECO:0007829|PDB:5H4U"
FT HELIX 152..156
FT /evidence="ECO:0007829|PDB:5H4U"
FT STRAND 163..165
FT /evidence="ECO:0007829|PDB:5H4U"
FT TURN 166..168
FT /evidence="ECO:0007829|PDB:5H4U"
FT HELIX 173..178
FT /evidence="ECO:0007829|PDB:5H4U"
FT HELIX 181..183
FT /evidence="ECO:0007829|PDB:5H4U"
FT HELIX 184..191
FT /evidence="ECO:0007829|PDB:5H4U"
FT STRAND 201..207
FT /evidence="ECO:0007829|PDB:5H4U"
FT HELIX 211..217
FT /evidence="ECO:0007829|PDB:5H4U"
SQ SEQUENCE 225 AA; 23795 MW; BF9402ED1CAFCAE2 CRC64;
MKVFVVLAAI VAIANGLTSG SGVTTRYWDC CKPSCSWGGK ASVTKPVRTC KANGNTTIDS
NTQSGCNGGS SYVCNDQQPF TQGNVGYGFA AASISGQPES QTCCACYEMT FTNTAISGQK
MIVQVTNTGS DLNGNHFDLM IPGGGVGIFN GCQSQWGAPS NGWGQRYGGI SSQSECNQLP
TSLRAGCNWR FGWFKNADNP SMKFTQVRCP TILTQKSQCV RTPGP
