LGK_LIPST
ID LGK_LIPST Reviewed; 439 AA.
AC B3VI55;
DT 23-FEB-2022, integrated into UniProtKB/Swiss-Prot.
DT 02-SEP-2008, sequence version 1.
DT 03-AUG-2022, entry version 35.
DE RecName: Full=Levoglucosan kinase {ECO:0000303|Ref.1};
DE Short=LGK {ECO:0000303|Ref.1};
DE EC=2.7.1.232 {ECO:0000269|PubMed:21719279, ECO:0000269|PubMed:26354439, ECO:0000269|Ref.1};
OS Lipomyces starkeyi (Oleaginous yeast).
OC Eukaryota; Fungi; Dikarya; Ascomycota; Saccharomycotina; Saccharomycetes;
OC Saccharomycetales; Lipomycetaceae; Lipomyces.
OX NCBI_TaxID=29829;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], FUNCTION, CATALYTIC ACTIVITY, AND
RP BIOPHYSICOCHEMICAL PROPERTIES.
RC STRAIN=YZ-215;
RX AGRICOLA=IND44243598; DOI=10.1007/s11274-009-0048-9;
RA Dai J., Yu Z., He Y., Zhang L., Du Y., Bai Z., Dong Z., Zhang H.;
RT "Cloning of a novel levoglucosan kinase gene from Lipomyces starkeyi and
RT its expression in Escherichia coli.";
RL World J. Microbiol. Biotechnol. 25:1589-1595(2009).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, AND BIOTECHNOLOGY.
RX PubMed=21719279; DOI=10.1016/j.biortech.2011.06.011;
RA Layton D.S., Ajjarapu A., Choi D.W., Jarboe L.R.;
RT "Engineering ethanologenic Escherichia coli for levoglucosan utilization.";
RL Bioresour. Technol. 102:8318-8322(2011).
RN [3] {ECO:0007744|PDB:4YH5, ECO:0007744|PDB:4ZFV, ECO:0007744|PDB:4ZLU, ECO:0007744|PDB:5BSB, ECO:0007744|PDB:5BVC}
RP X-RAY CRYSTALLOGRAPHY (1.50 ANGSTROMS) IN COMPLEX WITH ADP; MAGNESIUM AND
RP LEVOGLUCOSAN, FUNCTION, COFACTOR, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL
RP PROPERTIES, SUBUNIT, AND ACTIVITY REGULATION.
RX PubMed=26354439; DOI=10.1074/jbc.m115.674614;
RA Bacik J.P., Klesmith J.R., Whitehead T.A., Jarboe L.R., Unkefer C.J.,
RA Mark B.L., Michalczyk R.;
RT "Producing glucose 6-phosphate from cellulosic biomass: structural insights
RT into levoglucosan bioconversion.";
RL J. Biol. Chem. 290:26638-26648(2015).
RN [4] {ECO:0007744|PDB:5TKR}
RP X-RAY CRYSTALLOGRAPHY (1.80 ANGSTROMS) IN COMPLEX WITH ADP AND MAGNESIUM,
RP COFACTOR, SUBUNIT, AND MUTAGENESIS OF CYS-194.
RX PubMed=28196882; DOI=10.1073/pnas.1614437114;
RA Klesmith J.R., Bacik J.P., Wrenbeck E.E., Michalczyk R., Whitehead T.A.;
RT "Trade-offs between enzyme fitness and solubility illuminated by deep
RT mutational scanning.";
RL Proc. Natl. Acad. Sci. U.S.A. 114:2265-2270(2017).
CC -!- FUNCTION: Levoglucosan kinase that catalyzes the transfer of a
CC phosphate group from ATP to levoglucosan (1,6-anhydro-beta-D-
CC glucopyranose, LG) to yield glucose 6-phosphate in the presence of
CC magnesium ion and ATP (Ref.1, PubMed:21719279, PubMed:26354439). In
CC addition to the canonical kinase phosphotransfer reaction, the
CC conversion requires cleavage of the 1,6-anhydro ring to allow ATP-
CC dependent phosphorylation of the sugar O-6 atom (Ref.1,
CC PubMed:21719279, PubMed:26354439). {ECO:0000269|PubMed:21719279,
CC ECO:0000269|PubMed:26354439, ECO:0000269|Ref.1}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=ATP + H2O + levoglucosan = ADP + D-glucose 6-phosphate + H(+);
CC Xref=Rhea:RHEA:63428, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:30616, ChEBI:CHEBI:30997, ChEBI:CHEBI:61548,
CC ChEBI:CHEBI:456216; EC=2.7.1.232;
CC Evidence={ECO:0000269|PubMed:21719279, ECO:0000269|PubMed:26354439,
CC ECO:0000269|Ref.1};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:63429;
CC Evidence={ECO:0000269|PubMed:21719279, ECO:0000269|PubMed:26354439,
CC ECO:0000269|Ref.1};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:26354439, ECO:0000269|PubMed:28196882};
CC Note=Binds 2 Mg(2+) ions per subunit. {ECO:0000269|PubMed:26354439,
CC ECO:0000269|PubMed:28196882};
CC -!- ACTIVITY REGULATION: Tris(hydroxymethyl)aminomethane (Tris) is a
CC competitive inhibitor of the reaction. {ECO:0000269|PubMed:26354439}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=105.3 mM for levoglucosan {ECO:0000269|PubMed:26354439,
CC ECO:0000269|Ref.1};
CC KM=0.20 mM for ATP {ECO:0000269|Ref.1};
CC pH dependence:
CC Optimum pH is 9.0. {ECO:0000269|Ref.1};
CC Temperature dependence:
CC Optimum temperature is 30 degrees Celsius. {ECO:0000269|Ref.1};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:26354439,
CC ECO:0000269|PubMed:28196882}.
CC -!- BIOTECHNOLOGY: Levoglucosan is the major anhydrosugar compound
CC resulting from the degradation of cellulose during the fast pyrolysis
CC process of biomass and thus the most attractive fermentation substrate
CC in the bio-oil (PubMed:21719279). Engineered E.coli can use
CC levoglucosan as sole carbon source, but it also ferments levoglucosan
CC to ethanol (PubMed:21719279). {ECO:0000269|PubMed:21719279}.
CC -!- SIMILARITY: Belongs to the anhydro-N-acetylmuramic acid kinase family.
CC {ECO:0000305}.
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DR EMBL; EU751287; ACE79748.1; -; mRNA.
DR PDB; 4YH5; X-ray; 1.90 A; A/B=1-439.
DR PDB; 4ZFV; X-ray; 1.50 A; A/B=1-439.
DR PDB; 4ZLU; X-ray; 1.80 A; A/B=1-439.
DR PDB; 5BSB; X-ray; 1.85 A; A=1-439.
DR PDB; 5BVC; X-ray; 2.00 A; A=1-439.
DR PDB; 5TKR; X-ray; 1.80 A; A=1-439.
DR PDBsum; 4YH5; -.
DR PDBsum; 4ZFV; -.
DR PDBsum; 4ZLU; -.
DR PDBsum; 5BSB; -.
DR PDBsum; 5BVC; -.
DR PDBsum; 5TKR; -.
DR SMR; B3VI55; -.
DR KEGG; ag:ACE79748; -.
DR BioCyc; MetaCyc:MON-21029; -.
DR BRENDA; 2.7.1.232; 3042.
DR GO; GO:0005524; F:ATP binding; IEA:InterPro.
DR GO; GO:0016301; F:kinase activity; IEA:UniProtKB-KW.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0016773; F:phosphotransferase activity, alcohol group as acceptor; IEA:InterPro.
DR GO; GO:0006040; P:amino sugar metabolic process; IEA:InterPro.
DR GO; GO:0009254; P:peptidoglycan turnover; IEA:InterPro.
DR GO; GO:0016310; P:phosphorylation; IEA:UniProtKB-KW.
DR InterPro; IPR005338; Anhydro_N_Ac-Mur_kinase.
DR InterPro; IPR043129; ATPase_NBD.
DR PANTHER; PTHR30605; PTHR30605; 1.
DR Pfam; PF03702; AnmK; 1.
DR SUPFAM; SSF53067; SSF53067; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Kinase; Magnesium; Metal-binding; Nucleotide-binding;
KW Transferase.
FT CHAIN 1..439
FT /note="Levoglucosan kinase"
FT /id="PRO_0000455010"
FT BINDING 23..26
FT /ligand="ADP"
FT /ligand_id="ChEBI:CHEBI:456216"
FT /evidence="ECO:0000269|PubMed:26354439,
FT ECO:0000269|PubMed:28196882, ECO:0007744|PDB:4YH5,
FT ECO:0007744|PDB:4ZFV, ECO:0007744|PDB:4ZLU,
FT ECO:0007744|PDB:5BVC, ECO:0007744|PDB:5TKR"
FT BINDING 26
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:26354439,
FT ECO:0000269|PubMed:28196882, ECO:0007744|PDB:4YH5,
FT ECO:0007744|PDB:4ZFV, ECO:0007744|PDB:4ZLU,
FT ECO:0007744|PDB:5BVC, ECO:0007744|PDB:5TKR"
FT BINDING 189
FT /ligand="ADP"
FT /ligand_id="ChEBI:CHEBI:456216"
FT /evidence="ECO:0000269|PubMed:26354439,
FT ECO:0000269|PubMed:28196882, ECO:0007744|PDB:4YH5,
FT ECO:0007744|PDB:4ZFV, ECO:0007744|PDB:4ZLU,
FT ECO:0007744|PDB:5BVC, ECO:0007744|PDB:5TKR"
FT BINDING 192
FT /ligand="levoglucosan"
FT /ligand_id="ChEBI:CHEBI:30997"
FT /evidence="ECO:0000269|PubMed:26354439,
FT ECO:0007744|PDB:4ZLU, ECO:0007744|PDB:5BVC"
FT BINDING 212
FT /ligand="levoglucosan"
FT /ligand_id="ChEBI:CHEBI:30997"
FT /evidence="ECO:0000269|PubMed:26354439,
FT ECO:0007744|PDB:4ZLU, ECO:0007744|PDB:5BSB,
FT ECO:0007744|PDB:5BVC"
FT BINDING 217
FT /ligand="ADP"
FT /ligand_id="ChEBI:CHEBI:456216"
FT /evidence="ECO:0007744|PDB:4ZLU"
FT BINDING 221
FT /ligand="ADP"
FT /ligand_id="ChEBI:CHEBI:456216"
FT /evidence="ECO:0000269|PubMed:26354439,
FT ECO:0000269|PubMed:28196882, ECO:0007744|PDB:4YH5,
FT ECO:0007744|PDB:4ZFV, ECO:0007744|PDB:4ZLU,
FT ECO:0007744|PDB:5BVC, ECO:0007744|PDB:5TKR"
FT BINDING 234..237
FT /ligand="ADP"
FT /ligand_id="ChEBI:CHEBI:456216"
FT /evidence="ECO:0000269|PubMed:26354439,
FT ECO:0000269|PubMed:28196882, ECO:0007744|PDB:4YH5,
FT ECO:0007744|PDB:4ZFV, ECO:0007744|PDB:4ZLU,
FT ECO:0007744|PDB:5BVC, ECO:0007744|PDB:5TKR"
FT BINDING 328
FT /ligand="ADP"
FT /ligand_id="ChEBI:CHEBI:456216"
FT /evidence="ECO:0000269|PubMed:26354439,
FT ECO:0000269|PubMed:28196882, ECO:0007744|PDB:4YH5,
FT ECO:0007744|PDB:4ZFV, ECO:0007744|PDB:4ZLU,
FT ECO:0007744|PDB:5BVC, ECO:0007744|PDB:5TKR"
FT BINDING 362
FT /ligand="ADP"
FT /ligand_id="ChEBI:CHEBI:456216"
FT /evidence="ECO:0000269|PubMed:26354439,
FT ECO:0000269|PubMed:28196882, ECO:0007744|PDB:4YH5,
FT ECO:0007744|PDB:4ZFV, ECO:0007744|PDB:4ZLU,
FT ECO:0007744|PDB:5BVC, ECO:0007744|PDB:5TKR"
FT BINDING 362
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:26354439,
FT ECO:0000269|PubMed:28196882, ECO:0007744|PDB:4YH5,
FT ECO:0007744|PDB:4ZFV, ECO:0007744|PDB:4ZLU,
FT ECO:0007744|PDB:5BVC, ECO:0007744|PDB:5TKR"
FT MUTAGEN 194
FT /note="C->T: Leads to low solubility, probably by
FT introducing a potential glycosylation site near the active
FT site that results in a misfolded protein that would be
FT retained in the endoplasmic reticulum."
FT /evidence="ECO:0000269|PubMed:28196882"
FT STRAND 12..21
FT /evidence="ECO:0007829|PDB:4ZFV"
FT STRAND 26..39
FT /evidence="ECO:0007829|PDB:4ZFV"
FT STRAND 44..55
FT /evidence="ECO:0007829|PDB:4ZFV"
FT HELIX 58..70
FT /evidence="ECO:0007829|PDB:4ZFV"
FT HELIX 75..99
FT /evidence="ECO:0007829|PDB:4ZFV"
FT HELIX 104..106
FT /evidence="ECO:0007829|PDB:4ZFV"
FT STRAND 109..120
FT /evidence="ECO:0007829|PDB:4ZFV"
FT STRAND 130..135
FT /evidence="ECO:0007829|PDB:4ZFV"
FT HELIX 137..144
FT /evidence="ECO:0007829|PDB:4ZFV"
FT STRAND 148..150
FT /evidence="ECO:0007829|PDB:4ZFV"
FT HELIX 152..158
FT /evidence="ECO:0007829|PDB:4ZFV"
FT HELIX 168..175
FT /evidence="ECO:0007829|PDB:4ZFV"
FT STRAND 182..196
FT /evidence="ECO:0007829|PDB:4ZFV"
FT STRAND 209..217
FT /evidence="ECO:0007829|PDB:4ZFV"
FT HELIX 218..227
FT /evidence="ECO:0007829|PDB:4ZFV"
FT TURN 228..230
FT /evidence="ECO:0007829|PDB:4ZFV"
FT HELIX 236..238
FT /evidence="ECO:0007829|PDB:4ZFV"
FT HELIX 239..243
FT /evidence="ECO:0007829|PDB:4ZFV"
FT HELIX 248..254
FT /evidence="ECO:0007829|PDB:4ZFV"
FT HELIX 258..261
FT /evidence="ECO:0007829|PDB:4ZFV"
FT TURN 270..272
FT /evidence="ECO:0007829|PDB:4ZFV"
FT HELIX 275..288
FT /evidence="ECO:0007829|PDB:4ZFV"
FT HELIX 292..314
FT /evidence="ECO:0007829|PDB:4ZFV"
FT STRAND 322..327
FT /evidence="ECO:0007829|PDB:4ZFV"
FT HELIX 328..331
FT /evidence="ECO:0007829|PDB:4ZFV"
FT HELIX 333..342
FT /evidence="ECO:0007829|PDB:4ZFV"
FT STRAND 346..350
FT /evidence="ECO:0007829|PDB:4ZFV"
FT HELIX 351..354
FT /evidence="ECO:0007829|PDB:4ZFV"
FT HELIX 358..360
FT /evidence="ECO:0007829|PDB:4ZFV"
FT HELIX 361..375
FT /evidence="ECO:0007829|PDB:4ZFV"
FT STRAND 383..386
FT /evidence="ECO:0007829|PDB:4ZFV"
FT STRAND 394..397
FT /evidence="ECO:0007829|PDB:4ZFV"
FT HELIX 402..411
FT /evidence="ECO:0007829|PDB:4ZFV"
FT TURN 412..415
FT /evidence="ECO:0007829|PDB:4ZFV"
FT STRAND 425..429
FT /evidence="ECO:0007829|PDB:4ZFV"
FT STRAND 432..434
FT /evidence="ECO:0007829|PDB:4ZFV"
SQ SEQUENCE 439 AA; 48372 MW; 4D34653E7E026CB3 CRC64;
MPIATSTGDN VLDFTVLGLN SGTSMDGIDC ALCHFYQKTP DAPMEFELLE YGEVPLAQPI
KQRVMRMILE DTTSPSELSE VNVILGEHFA DAVRQFAAER NVDLSTIDAI ASHGQTIWLL
SMPEEGQVKS ALTMAEGAIL ASRTGITSIT DFRISDQAAG RQGAPLIAFF DALLLHHPTK
LRACQNIGGI ANVCFIPPDV DGRRTDEYYD FDTGPGNVFI DAVVRHFTNG EQEYDKDGAM
GKRGKVDQEL VDDFLKMPYF QLDPPKTTGR EVFRDTLAHD LIRRAEAKGL SPDDIVATTT
RITAQAIVDH YRRYAPSQEI DEIFMCGGGA YNPNIVEFIQ QSYPNTKIMM LDEAGVPAGA
KEAITFAWQG MEALVGRSIP VPTRVETRQH YVLGKVSPGL NYRSVMKKGM AFGGDAQQLP
WVSEMIVKKK GKVITNNWA
