MANA_CRYAT
ID MANA_CRYAT Reviewed; 382 AA.
AC B4XC07;
DT 12-AUG-2020, integrated into UniProtKB/Swiss-Prot.
DT 23-SEP-2008, sequence version 1.
DT 03-AUG-2022, entry version 57.
DE RecName: Full=Mannan endo-1,4-beta-mannosidase {ECO:0000305};
DE EC=3.2.1.78 {ECO:0000269|PubMed:18579426, ECO:0000269|PubMed:22333528, ECO:0000269|PubMed:25082572, ECO:0000312|EMBL:ABV68808.1};
DE AltName: Full=Beta-mannanase {ECO:0000303|PubMed:18579426};
DE AltName: Full=CaMan {ECO:0000303|PubMed:18579426, ECO:0000303|PubMed:22333528, ECO:0000303|PubMed:23989150, ECO:0000303|PubMed:25082572};
DE AltName: Full=Endo-beta-1,4-D-mannanase {ECO:0000303|PubMed:18579426, ECO:0000303|PubMed:25082572};
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:ABV68808.1};
RN [1] {ECO:0000312|EMBL:ABV68808.1}
RP NUCLEOTIDE SEQUENCE [MRNA], FUNCTION, SUBSTRATE SPECIFICITY, CATALYTIC
RP ACTIVITY, ACTIVITY REGULATION, BIOPHYSICOCHEMICAL PROPERTIES, AND
RP BIOTECHNOLOGY.
RX PubMed=18579426; DOI=10.1016/j.cbpb.2008.05.005;
RA Song J.M., Nam K.-W., Kang S.G., Kim C.-G., Kwon S.-T., Lee Y.-H.;
RT "Molecular cloning and characterization of a novel cold-active beta-1,4-D-
RT mannanase from the Antarctic springtail, Cryptopygus antarcticus.";
RL Comp. Biochem. Physiol. 151B:32-40(2008).
RN [2]
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 [3]
RP CRYSTALLIZATION, AND SUBUNIT.
RX PubMed=23989150; DOI=10.1107/s1744309113020538;
RA Kim M.K., An Y.J., Jeong C.S., Song J.M., Kang M.H., Lee Y.H., Cha S.S.;
RT "Expression at 279 K, purification, crystallization and preliminary X-ray
RT crystallographic analysis of a novel cold-active beta-1,4-D-mannanase from
RT the Antarctic springtail Cryptopygus antarcticus.";
RL Acta Crystallogr. F 69:1007-1010(2013).
RN [4] {ECO:0007744|PDB:4OOU, ECO:0007744|PDB:4OOZ}
RP X-RAY CRYSTALLOGRAPHY (2.36 ANGSTROMS) AND IN COMPLEX WITH MANNOPENTAOSE,
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT,
RP DISULFIDE BONDS, AND MUTAGENESIS OF TRP-243; HIS-284; TRP-341 AND
RP 346-GLY--SER-350.
RX PubMed=25082572; DOI=10.1002/prot.24655;
RA Kim M.K., An Y.J., Song J.M., Jeong C.S., Kang M.H., Kwon K.K., Lee Y.H.,
RA Cha S.S.;
RT "Structure-based investigation into the functional roles of the extended
RT loop and substrate-recognition sites in an endo-beta-1,4-D-mannanase from
RT the Antarctic springtail, Cryptopygus antarcticus.";
RL Proteins 82:3217-3223(2014).
CC -!- FUNCTION: Hydrolyzes 1,4-beta linked polysaccharide backbones of
CC mannans. Has high activity toward locust bean gum (PubMed:18579426,
CC PubMed:25082572). Also active toward konjac and beta-1,4-mannan.
CC Hydrolyzes mannotetraose (M4) and mannopentaose (M5) to mannobiose (M2)
CC and mannotriose (M3) with a little production of mannose (M1).
CC Hydrolyzes beta-1,4-mannan to M2, M3 and M4. Hardly hydrolyzes M2 and
CC M3. Does not hydrolyze p-nitrophenyl-beta-D-mannopyranoside, gua-gum,
CC carboxymethyl cellulose, soluble starch or laminarin (PubMed:18579426).
CC {ECO:0000269|PubMed:18579426, ECO:0000269|PubMed:25082572}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Random hydrolysis of (1->4)-beta-D-mannosidic linkages in
CC mannans, galactomannans and glucomannans.; EC=3.2.1.78;
CC Evidence={ECO:0000269|PubMed:18579426, ECO:0000269|PubMed:22333528,
CC ECO:0000269|PubMed:25082572};
CC -!- ACTIVITY REGULATION: Activated particularly by Ca(2+) and Zn(2+), and
CC to a lesser extent by Na(+), K(+), Mg(2+) and Cu(2+). Activation effect
CC of the divalent metal ions Ca(2+), Zn(2+), Mg(2+) and Cu(2+) is reduced
CC significantly by the addition of EDTA. Strongly inhibited by Mn(2+),
CC Hg(2+) and Ag(+). {ECO:0000269|PubMed:18579426}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=5.61 mg/ml for 0.5% locust bean gum (at pH 3.5 and 10 degrees
CC Celsius in the presence of 50mM sodium citrate)
CC {ECO:0000269|PubMed:25082572};
CC Vmax=2477.22 mol/min/mg enzyme with 0.5% locust bean gum as substrate
CC (at pH 3.5 and 10 degrees Celsius in the presence of 50mM sodium
CC citrate) {ECO:0000269|PubMed:25082572};
CC Note=kcat is 1659.90 sec(-1) for 0.5% locust bean gum (at pH 3.5 and
CC 10 degrees Celsius in the presence of 50mM sodium citrate).
CC {ECO:0000269|PubMed:25082572};
CC pH dependence:
CC Optimum pH is 3.5. Maintains more than 50% of the maximum activity at
CC pH 2.5-6.0. Not active at a pH above 7.0.
CC {ECO:0000269|PubMed:18579426};
CC Temperature dependence:
CC Optimum temperature is 30 degrees Celsius. Retains approximately 20-
CC 40% of its maximum activity even at 0-5 degrees Celsius. Drastic loss
CC of activity at temperatures above 45 degrees Celsius, at which the
CC half-life of the enzyme activity is less than 10 min.
CC {ECO:0000269|PubMed:18579426};
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:23989150,
CC ECO:0000269|PubMed:25082572}.
CC -!- SUBCELLULAR LOCATION: Secreted {ECO:0000305}.
CC -!- BIOTECHNOLOGY: Potential use in mannan-hydrolysis industries which need
CC especially low temperature and an acidic condition such as the food and
CC the animal feed industries. {ECO:0000305|PubMed:18579426}.
CC -!- SIMILARITY: Belongs to the glycosyl hydrolase 5 (cellulase A) family.
CC {ECO:0000305}.
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DR EMBL; EU021047; ABV68808.1; -; mRNA.
DR PDB; 4OOU; X-ray; 2.36 A; A/B=1-382.
DR PDB; 4OOZ; X-ray; 2.60 A; A/B=1-382.
DR PDBsum; 4OOU; -.
DR PDBsum; 4OOZ; -.
DR AlphaFoldDB; B4XC07; -.
DR SMR; B4XC07; -.
DR CAZy; GH5; Glycoside Hydrolase Family 5.
DR BRENDA; 3.2.1.78; 11159.
DR GO; GO:0005576; C:extracellular region; IC:UniProtKB.
DR GO; GO:0016985; F:mannan endo-1,4-beta-mannosidase activity; IDA:UniProtKB.
DR GO; GO:0070492; F:oligosaccharide binding; IDA:UniProtKB.
DR GO; GO:0030247; F:polysaccharide binding; IC:UniProtKB.
DR GO; GO:0046355; P:mannan catabolic process; IDA:UniProtKB.
DR InterPro; IPR017853; Glycoside_hydrolase_SF.
DR SUPFAM; SSF51445; SSF51445; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Carbohydrate metabolism; Disulfide bond; Glycosidase;
KW Hydrolase; Polysaccharide degradation; Secreted; Signal.
FT SIGNAL 1..19
FT /evidence="ECO:0000255"
FT CHAIN 20..382
FT /note="Mannan endo-1,4-beta-mannosidase"
FT /evidence="ECO:0000255"
FT /id="PRO_5002828363"
FT REGION 346..350
FT /note="Involved in stabilization of the transition state"
FT /evidence="ECO:0000305|PubMed:25082572"
FT ACT_SITE 181
FT /note="Proton donor/acceptor"
FT /evidence="ECO:0000250|UniProtKB:Q99036"
FT ACT_SITE 312
FT /note="Nucleophile"
FT /evidence="ECO:0000250|UniProtKB:Q99036"
FT BINDING 83
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:25082572,
FT ECO:0007744|PDB:4OOZ"
FT BINDING 144
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:25082572,
FT ECO:0007744|PDB:4OOZ"
FT BINDING 147..151
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:25082572,
FT ECO:0007744|PDB:4OOZ"
FT BINDING 180
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:25082572,
FT ECO:0007744|PDB:4OOZ"
FT BINDING 187
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:25082572,
FT ECO:0007744|PDB:4OOZ"
FT BINDING 204
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:25082572,
FT ECO:0007744|PDB:4OOZ"
FT BINDING 208
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:25082572,
FT ECO:0007744|PDB:4OOZ"
FT BINDING 243
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:25082572,
FT ECO:0007744|PDB:4OOZ"
FT BINDING 282
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:25082572,
FT ECO:0007744|PDB:4OOZ"
FT BINDING 284
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:25082572,
FT ECO:0007744|PDB:4OOZ"
FT BINDING 341
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:25082572,
FT ECO:0007744|PDB:4OOZ"
FT BINDING 348
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:25082572,
FT ECO:0007744|PDB:4OOZ"
FT DISULFID 195..262
FT /evidence="ECO:0000269|PubMed:25082572,
FT ECO:0007744|PDB:4OOU, ECO:0007744|PDB:4OOZ"
FT DISULFID 317..349
FT /evidence="ECO:0000269|PubMed:25082572,
FT ECO:0007744|PDB:4OOU, ECO:0007744|PDB:4OOZ"
FT MUTAGEN 243
FT /note="W->F: 2.5-fold reduction in substrate affinity and
FT minor reduction in substrate turnover compared to the wild-
FT type."
FT /evidence="ECO:0000269|PubMed:25082572"
FT MUTAGEN 284
FT /note="H->A: Slight reduction in substrate affinity and
FT minor reduction in substrate turnover compared to the wild-
FT type."
FT /evidence="ECO:0000269|PubMed:25082572"
FT MUTAGEN 284
FT /note="H->W: About 1.4-fold increase in substrate affinity,
FT but 1.5-fold reduction in substrate turnover compared to
FT the wild-type."
FT /evidence="ECO:0000269|PubMed:25082572"
FT MUTAGEN 341
FT /note="W->F: 2-fold reduction in substrate affinity and 10-
FT fold reduction in substrate turnover compared to the wild-
FT type."
FT /evidence="ECO:0000269|PubMed:25082572"
FT MUTAGEN 346..350
FT /note="Missing: Optimum temperature is increased by 5
FT degrees Celsius compared to the wild-type. Exhibits
FT approximately half of its Vmax at 0-30 degrees Celsius. No
FT significant difference in substrate affinity, but nearly 2-
FT fold reduction in substrate turnover compared to the wild-
FT type. Requires 1.8 kcalmol(-1) higher activation enthalpy
FT than the wild-type."
FT /evidence="ECO:0000269|PubMed:25082572"
FT STRAND 24..26
FT /evidence="ECO:0007829|PDB:4OOU"
FT STRAND 29..32
FT /evidence="ECO:0007829|PDB:4OOU"
FT STRAND 35..37
FT /evidence="ECO:0007829|PDB:4OOU"
FT STRAND 39..43
FT /evidence="ECO:0007829|PDB:4OOU"
FT STRAND 52..54
FT /evidence="ECO:0007829|PDB:4OOU"
FT HELIX 58..74
FT /evidence="ECO:0007829|PDB:4OOU"
FT STRAND 79..86
FT /evidence="ECO:0007829|PDB:4OOU"
FT STRAND 89..94
FT /evidence="ECO:0007829|PDB:4OOU"
FT STRAND 100..103
FT /evidence="ECO:0007829|PDB:4OOU"
FT HELIX 109..122
FT /evidence="ECO:0007829|PDB:4OOU"
FT STRAND 126..135
FT /evidence="ECO:0007829|PDB:4OOU"
FT HELIX 140..145
FT /evidence="ECO:0007829|PDB:4OOU"
FT HELIX 149..158
FT /evidence="ECO:0007829|PDB:4OOU"
FT HELIX 160..167
FT /evidence="ECO:0007829|PDB:4OOU"
FT STRAND 173..180
FT /evidence="ECO:0007829|PDB:4OOU"
FT HELIX 182..184
FT /evidence="ECO:0007829|PDB:4OOU"
FT TURN 194..196
FT /evidence="ECO:0007829|PDB:4OOU"
FT TURN 199..209
FT /evidence="ECO:0007829|PDB:4OOU"
FT HELIX 215..232
FT /evidence="ECO:0007829|PDB:4OOU"
FT STRAND 238..244
FT /evidence="ECO:0007829|PDB:4OOU"
FT HELIX 245..247
FT /evidence="ECO:0007829|PDB:4OOU"
FT HELIX 251..253
FT /evidence="ECO:0007829|PDB:4OOU"
FT HELIX 260..267
FT /evidence="ECO:0007829|PDB:4OOU"
FT STRAND 275..281
FT /evidence="ECO:0007829|PDB:4OOU"
FT HELIX 293..295
FT /evidence="ECO:0007829|PDB:4OOU"
FT HELIX 299..301
FT /evidence="ECO:0007829|PDB:4OOU"
FT STRAND 308..314
FT /evidence="ECO:0007829|PDB:4OOU"
FT STRAND 317..320
FT /evidence="ECO:0007829|PDB:4OOU"
FT HELIX 323..332
FT /evidence="ECO:0007829|PDB:4OOU"
FT STRAND 336..341
FT /evidence="ECO:0007829|PDB:4OOU"
FT STRAND 347..349
FT /evidence="ECO:0007829|PDB:4OOZ"
FT HELIX 353..361
FT /evidence="ECO:0007829|PDB:4OOU"
FT TURN 362..365
FT /evidence="ECO:0007829|PDB:4OOU"
SQ SEQUENCE 382 AA; 41833 MW; 9521BA15DED63968 CRC64;
MVKLFSFLLL VWVASPAFSS EFLKASGSNF YYGGQKVFLS GVNFAWRSYG SDFGNGQYAS
NGPALKDWIN KVKASGGNTA RVWVHVEGQV SPAFDSHGFV TSTDSKKTLI NDLSDLLDYA
NGQNVFLILV LFNGALQNNS NVQNLFWDES KLNSYINNAL TPMVNALKSK PSLAAWEVLN
EPEGTLQPGS DQNSCYDTST LAAQGAGWGG KKFPMKQILK TINWISSAIH NADSKALVTV
GSWSELTQTD SFGYRNHYKD SCLTGAGGKS NGIINFYQMH TYSHSGKWNQ NAPFKVNRWA
YNVNDKPLLI GEFASVCSQN EGIQNLYKYA YNNGYNGALT WQFNSGGDCS DTYSNQMYGM
QALKGQNDQS GGKGGMVSVN IN
