AGL31A_CANTS
  Name and origin · Biochemical properties · Enzyme annotation · Literature · Protein features · Sequences · Cross-refs · Entry history
Name and origin
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Gene Name
Name: agl31A
Other given name:
Protein names
Common name: alpha-glucosidase
EC systematic name: 4-alpha-D-glucan glucohydrolase
Other name:
Organism
Species: Candida tsukubaensis
Strain: CBS 6389
Taxonomic identifier: 5483
Taxonomic lineage: Eukaryota › Fungi › Dikarya › Basidiomycota › Ustilaginomycotina › Ustilaginomycetes › Ustilaginales › Ustilaginaceae › mitosporic Ustilaginaceae › Pseudozyma
Enzyme activity
Biochemical properties
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General properties
Expression host
Saccharomyces cerevisiae AH22
Substrate
maltose
Assay
Glucose oxidase-peroxidase assay.
Temperature Optimum(°C)
58
Temperature Stability(°C)
pH Optimum
4.2-4.6
pH Stability
3.9-4.9
Kinetic properties
Host Substrate Activity assay conditions Specific activity Relative activity(%) Assay Reference
Activity assay
Saccharomyces cerevisiae AH22 maltose citrate-phosphate buffer, pH 5.4, 50°C. active 100 Glucose oxidase-peroxidase assay.
Saccharomyces cerevisiae AH22 maltotriose citrate-phosphate buffer, pH 5.4, 50°C. active 59 Glucose oxidase-peroxidase assay.
Saccharomyces cerevisiae AH22 maltotetraose citrate-phosphate buffer, pH 5.4, 50°C. active 48 Glucose oxidase-peroxidase assay.
Saccharomyces cerevisiae AH22 sucrose citrate-phosphate buffer, pH 5.4, 50°C. active 55 Glucose oxidase-peroxidase assay.
Saccharomyces cerevisiae AH22 panose citrate-phosphate buffer, pH 5.4, 50°C. active 55 Glucose oxidase-peroxidase assay.
Saccharomyces cerevisiae AH22 isomaltose citrate-phosphate buffer, pH 5.4, 50°C. active 35 Glucose oxidase-peroxidase assay.
Saccharomyces cerevisiae AH22 phenyl alpha-D-glucose citrate-phosphate buffer, pH 5.4, 50°C. active 18 Glucose oxidase-peroxidase assay.
 
Host Substrate Activity assay conditions Product formed Product analysis Reference
Product analysis
 
Host Substrate Kinetic assay conditions Km Kcat(s-1) Vmax Reference
Kinetic assay
 
Other features
- Composed of two polypeptide chains.
- The recombinant enzyme is not active on potato starch, corn starch, amylopectin, pullulan, trehalose or alpha-cyclodextrin.
- Broad substrate specificity.
Enzyme annotation
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Enzyme
   Enzyme commission
3.2.1.20, 3.2.1.21; 4-alpha-D-glucan glucohydrolase
Gene Ontology
GO ID Term Evidence Reference
   Biological process
GO:0000025 maltose catabolic process IDA
GO:0005987 sucrose catabolic process IDA
   Molecular function
GO:0004558 alpha-glucosidase activity IDA
   Cellular component
GO:0005576 extracellular region IDA
Literature
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[1] Molecular cloning and characterization of a Candida tsukubaensis alpha-glucosidase gene in the yeast Saccharomyces cerevisiae.  
Author Kinsella BT, Larkin A, Bolton M and Cantwell BA
Journal Curr. Genet. 1991 Jul;20(1-2):45-52.
Address Guinness Brewing Worldwide Research Centre, St. James's Gate, Dublin, Ireland.
Abstract The molecular cloning of an alpha-glucosidase gene isolated from a Candida tsukubaensis (CBS 6389) genomic library in Saccharomyces cervisiae is reported. The cloned gene is contained within a 6.2 kb Sau3A DNA fragment and directs the synthesis and secretion of an amylolytic enzyme into the extracellular medium of the recombinant host, S. cerevisiae. The cloned enzyme was found to have an unusually broad substrate specificity and is capable of hydrolysing alpha-1,2, alpha-1,3, alpha-1,4 and alpha-1,6 linked, as well as aryl and alkyl, D-glucosides. On the basis of its substrate specificity profile, the cloned enzyme was classified as an alpha-glucosidase (E.C. 3.2.1.20). It has a pH optimum in the range 4.2-4.6, a temperature optimum of 58 degrees C and is readily inactivated at pasteurization temperature (60 degrees C). Southern blot analysis failed to reveal any homology between the cloned gene and genomic DNA isolated from other well characterized amylolytic yeasts. A rapid plate-assay, based on the utilization of a chromogenic substrate X-alpha-D-glucoside to detect the expression of the cloned alpha-glucosidase in S. cerevisiae transformants, was developed.
[2] Primary structure and processing of the Candida tsukubaensis alpha-glucosidase. Homology with the rabbit intestinal sucrase-isomaltase complex and human lysosomal alpha-glucosidase.  
Author Kinsella BT, Hogan S, Larkin A and Cantwell BA
Journal Eur. J. Biochem. 1991 Dec 5;202(2):657-64.
Address Guinness Brewing Worldwide Research Centre, St. James's Gate Brewery, Dublin, Ireland.
Abstract The nucleotide sequence of a 4.39-kb DNA fragment encoding the alpha-glucosidase gene of Candida tsukubaensis is reported. The cloned gene contains a major open reading frame (ORF 1) which encodes the alpha-glucosidase as a single precursor polypeptide of 1070 amino acids with a predicted molecular mass of 119 kDa. N-terminal amino acid sequence analysis of the individual subunits of the purified enzyme, expressed in the recombinant host Saccharomyces cerevisiae, confirmed that the alpha-glucosidase precursor is proteolytically processed by removal of an N-terminal signal peptide to yield the two peptide subunits 1 and 2, of molecular masses 63-65 kDa and 50-52 kDa, respectively. Both subunits are secreted by the heterologous host S. cerevisiae in a glycosylated form. Coincident with its efficient expression in the heterologous host, the C. tsukubaensis alpha-glucosidase gene contains many of the canonical features of highly expressed S. cerevisiae genes. There is considerable sequence similarity between C. tsukubaensis alpha-glucosidase, the rabbit sucrase-isomaltase complex (proSI) and human lysosomal acid alpha-glucosidase. The cloned DNA fragment from C. tsukubaensis contains a second open reading frame (ORF 2) which has the capacity to encode a polypeptide of 170 amino acids. The function and identity of the polypeptide encoded by ORF 2 is not known.
Protein features
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Signal Peptide (Predicted) SP(25aa)
N-Terminal (Experimental) 35aa
Structure PMID Reference
Sequence PMID Reference
CBM CBM
Glycosylation N-glycosylation
Other Domains
Domain Order
CAZy family GH31
Sequences
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Length 1070aa
Molecular Weight in kDa (experimental) 70 to 80 (recombinant enzyme; SDS-PAGE), 65 (recombinant enzyme subunit 1; deglycosylated with EndoH; SDS-PAGE), 50 to 52 (recombinant enzyme subunit 2; deglycosylated with EndoH; SDS-PAGE)
Molecular Weight in kDa (predicted) 116.07
Protein Sequence P29064
MRSIKAASLT PLLAALFTTL SSTLALPSSV WEHQLETNVL ALRDTNNNGS SSTISPSFDV TKCPGYKLVG QPQQSQHGFT AQLSLAGDAC NAYGVDIANL TLSVVYEKQH QLHVHIYDTA KQQYQLPNGL IFDRPGDNPA DIQNGSTADQ SDLVFHHTAE NGTQSGNGGW AFWIARKSSG DVIFDTRASN IPTYNDGLSS VSSNTKRNTT AMPAHEMVFE NQYLQISSAL PTGANIYGLG EYVTGSFRRN PDETLQPFFT LDAGTPVDSN MYGYHPIYTE ARRGSDGKLR THSVHLQNTA GMDVLLRRGV IQYRAIGGTL DFRFFSGDQP ASSSSSSSGN DKAVATVKNS PNTAIQQYVN FIGNPVIHPY WSYGFHLCRW GYNNVSETQA VIDAMRQNNI PLEVQWNDID YLQEFRDFTT DPQRFPQKEF AAMIAKLKDN HQHYIPIIDM AIPKAPTNDT DVYYPGTRGD ELDVFIKNRN GSQYIGEVWP GYTNFVDQQA ENAGKWWTEA IRNFSEIVDF SGIWLDMNEP SSFVIGNAAG PETNLSNTPA YTAATSVAGW PQGYNNLTWG TSGNITVNGS YTYQQGPVQN NDGSKQRRSL LLSRDEDVLV QRDINVNGGN GDKFGPEDPN YQYANSSQRY LSNPPYAIHN GIHISETPLN VNLDKKTVAM EAVGVDGQRA FYDVHNLDGT LEEQHFYNAL RDIRPQERPF LISRSTYPGA GKFTGHWLGD NYALWTILPG EEAYKAGAGM AQSIDGVLQF QIFGIHLIGA DICGFNRNSD EELCNRWMML GAFLPFMRNH NTIGAIAQEP FRWDSVANAS RIAINKRYEI LPSLYSHMAQ SAESGEPAVR ALWYEFDEVF EQTKDYAHQF LFGDDLLVSP VLEPNVTQIK ALFPNAGGKW RNVFSYEALD VEYNKNVTVD AALSTINVHL RPGKVLLTHS KPAYTVYETA QSPYGLIVNL NDQGEAKQTF YLDDGMTPAP TPNSTLTVSA GNNSVNGSIE GEYKAQQNLT YVVVLDVKQK PTQVMMGGNK TEFSWDQQKT LLNVTGLNAD LNGSGRFRGL RLELSLLCED
DNA Sequence X56024
GGATCCCCTG CTTAGCTTCT CTCCGCAAAG CCCCTGCTCA GCCTAGCCGT ACAAGTACCT TTCCATAAAT AGACGGCCAC TCTTCCTTCG TCAAACAATC TTTTCTTTCC ACATCAACAT CAACACAAGG TCTTCTTTCG CAAGCTCAGA AAGGCTTCCT TCCCCCAAAA TGCGTTCCAT CAAGGCGGCT TCATTGACGC CACTCTTGGC CGCTCTCTTC ACCACTCTCT CGTCTACTCT CGCCCTCCCT TCATCCGTCT GGGAACACCA GCTCGAGACC AACGTTCTCG CCCTACGAGA CACCAACAAC AATGGCTCCT CGTCCACCAT CTCCCCTTCC TTCGACGTGA CCAAGTGTCC AGGCTACAAG CTCGTCGGCC AACCTCAGCA ATCTCAGCAT GGCTTCACCG CCCAGCTCTC CCTCGCTGGT GACGCTTGCA ACGCTTATGG TGTCGACATC GCCAACTTGA CCCTCTCGGT CGTTTATGAG AAGCAGCATC AGCTTCATGT CCACATCTAC GACACCGCCA AGCAGCAATA CCAGCTCCCC AACGGTCTCA TCTTTGATCG TCCCGGCGAC AACCCTGCTG ACATTCAAAA TGGTTCTACT GCTGACCAGA GCGATCTCGT CTTCCACCAC ACTGCCGAGA ACGGTACACA GTCCGGCAAC GGCGGATGGG CGTTCTGGAT CGCACGCAAG TCGTCGGGCG ATGTGATCTT TGACACTCGC GCGAGCAACA TCCCCACCTA CAACGACGGA CTTAGCAGTG TCTCGTCCAA CACAAAACGC AACACCACGG CCATGCCGGC TCACGAAATG GTCTTTGAGA ACCAGTACCT TCAGATCTCC TCTGCCTTGC CAACGGGTGC CAACATTTAC GGTCTCGGAG AGTACGTGAC CGGTAGCTTC AGGAGGAACC CGGATGAAAC GCTGCAGCCG TTCTTCACTC TCGATGCTGG TACACCTGTT GATTCCAACA TGTACGGCTA CCACCCAATC TACACCGAAG CTAGAAGAGG TAGCGATGGT AAGCTTAGGA CGCACTCTGT TCACCTGCAA AACACGGCTG GTATGGATGT GCTTCTGCGT AGGGGTGTCA TTCAGTACAG GGCGATTGGC GGTACGCTCG ACTTCCGATT CTTTTCTGGT GACCAGCCTG CTTCCTCTTC TTCGTCTTCG AGCGGAAACG ATAAGGCAGT TGCAACCGTA AAGAACAGCC CGAACACTGC CATTCAGCAG TACGTCAACT TCATTGGCAA CCCCGTCATT CACCCTTACT GGTCGTATGG CTTCCACCTC TGCCGATGGG GCTACAACAA CGTTTCCGAG ACCCAAGCCG TCATCGACGC CATGCGACAG AACAACATCC CCCTCGAAGT GCAATGGAAC GATATCGACT ACTTGCAAGA GTTCCGTGAC TTTACCACGG ACCCGCAACG ATTCCCTCAG AAAGAGTTTG CTGCTATGAT TGCCAAGCTC AAGGATAACC ACCAGCACTA CATTCCTATC ATCGACATGG CTATTCCTAA GGCTCCGACG AACGACACCG ACGTCTACTA TCCTGGTACT CGTGGCGATG AGCTCGACGT CTTCATCAAG AATCGCAACG GCTCGCAATA CATCGGTGAA GTTTGGCCCG GTTACACCAA CTTTGTCGAC CAACAGGCCG AGAATGCGGG CAAGTGGTGG ACCGAGGCGA TCCGCAACTT TAGCGAGATT GTCGACTTCT CAGGTATCTG GCTGGATATG AACGAGCCTT CGAGTTTCGT TATCGGTAAT GCTGCTGGGC CGGAGACGAA TTTGTCGAAC ACGCCAGCCT ATACTGCGGC GACGAGCGTT GCTGGATGGC CGCAGGGTTA CAACAACCTC ACTTGGGGCA CTTCGGGTAA TATCACCGTC AACGGCAGTT ACACTTACCA GCAAGGACCT GTTCAGAACA ACGACGGCTC GAAGCAGCGT CGTTCGCTTC TGCTTTCGCG CGACGAAGAT GTTCTCGTCC AGCGTGACAT CAACGTCAAC GGTGGCAATG GCGACAAGTT TGGCCCCGAA GATCCCAACT ATCAATACGC TAACTCTTCG CAGCGATACC TCTCCAACCC TCCCTACGCC ATCCACAACG GTATCCACAT CAGCGAAACC CCCCTTAACG TCAACTTGGA TAAGAAGACC GTTGCTATGG AAGCTGTTGG TGTCGACGGT CAACGCGCTT TCTACGACGT TCACAATCTT GATGGCACCC TGGAGGAGCA ACACTTCTAC AACGCTCTTC GCGATATTCG CCCGCAAGAA CGACCGTTCC TCATCTCACG TTCTACCTAC CCTGGTGCAG GAAAGTTCAC CGGTCATTGG TTGGGTGACA ACTATGCGCT TTGGACCATC TTGCCCGGCG AAGAGGCGTA CAAAGCTGGC GCTGGTATGG CGCAGTCGAT CGACGGTGTA CTCCAGTTCC AGATCTTTGG CATCCACCTT ATTGGAGCCG ACATTTGCGG ATTTAACCGT AACAGTGACG AAGAGCTGTG CAATCGTTGG ATGATGCTTG GTGCATTCTT GCCGTTCATG CGCAACCACA ATACGATTGG TGCGATTGCT CAGGAGCCTT TCCGATGGGA CAGTGTTGCC AACGCGAGCA GGATCGCGAT CAACAAGAGG TACGAGATCT TGCCTTCGCT GTATTCGCAT ATGGCTCAGA GCGCCGAATC GGGCGAACCA GCGGTGAGGG CGCTGTGGTA CGAGTTCGAT GAGGTTTTCG AACAGACCAA GGATTACGCT CATCAGTTCT TGTTCGGAGA CGACTTGTTG GTCAGTCCAG TGTTGGAGCC CAATGTGACG CAGATCAAGG CGTTGTTCCC TAACGCCGGC GGAAAGTGGA GGAATGTTTT CAGCTACGAA GCGCTCGACG TTGAATACAA TAAGAACGTA ACCGTTGATG CGGCGCTTTC GACGATCAAC GTCCACCTTC GACCAGGAAA GGTTCTCCTC ACCCACTCCA AACCCGCGTA CACGGTCTAC GAAACCGCTC AAAGCCCCTA CGGCCTCATT GTCAACTTGA ACGACCAAGG AGAGGCAAAG CAAACCTTCT ACCTCGATGA CGGTATGACA CCCGCCCCCA CCCCTAACTC GACACTTACC GTAAGCGCAG GTAACAACTC GGTCAACGGA AGCATCGAAG GAGAGTACAA GGCGCAGCAA AACTTGACCT ACGTTGTCGT GCTCGATGTC AAGCAGAAGC CAACCCAGGT CATGATGGGT GGTAACAAGA CCGAGTTCAG TTGGGATCAG CAGAAGACGT TGCTCAACGT GACCGGCTTG AACGCTGATT TGAACGGAAG TGGTCGCTTT CGTGGTCTTA GACTCGAATT ATCCCTTCTT TGCGAGGATT GAAATGCAAA CTTTGGTTGC TTTCCTGAGT TCGACGGAGT GTGATATTGC GCCGTGTGAT AGCGGAGTGT GTGATTATAG AGTTGCAACG CGTGGTGATG AGGTGAAAGA CACACCTCAT CCTCTGAGGC TTCCGAATCG ACGCGGACAT CTGCTCAGCG AGAGTGAGTG AGCTCAGTCC ACTTGCCCAC TAGCCCAAAC CTTGTTTGGT GATGAGATCC TGATAGGACG GAGACAAGCC GCGAGTGGCG ATAAGGTGCG GAAAGAACGC ATTGCTTCTC TATCCAGCGC GCTGATGAGT CTCCTGACAG GACGCCTTTT GCCATGATCC CTTGTACATA TAAAGATAGG CAAAGAGACG TTCGCCTCCT TTCCATCTTC GTACCATCAT CCTTTCTCCC ATCTCCCCTC ATCATCATCC TTCCTCCCTT CCCTCTCACG GCACTCTCCT CGGCCACTCC AAGACTATCT CCTCACCATG TCGGCTCCTC AGACCACCGG CGACCGTGCT GTCATCAAGC TTCTCAGCGA TGCACTCAAA GCTCTCAACC TTCAGAATCT AGCCCAGACA CAGTTTCAGG AGGAGGTCAT GCCAGCCACC GCTCCCTCAA CTGACCCTGC TGTCCCCAAA GACGCACAGG AGGCTGACGA ACCCTCGATC GATGCGCTCA AGAGCAAGAT TGCAGGTTAC GAACGCATGT TCGCCTTCAA CTGCCAATTG CTTGGCTATT TGATGGAGGA AATCGACGGA GGCTCCCCTG ATTGGATGAA GGTTCCTTGC TACCTTTTCT TGATCATCCA ATCTTCGACT CTCACTCCTA TTGCAACAGG TGCGATCCCC AAAGGAATGG TCAAGGATGT CATTAGCCGC TTGCCTCCTA AGTATAAGGA ACGTTGCCAT GCAGCCGACA AGAATGAAGA TTGGCAGGCC TTGATGTTGG GTATCACAGA TCTCATGGAT GTTGTAAGTC TTTCTCTGCC ACCATCTTGA AATCCCTTTC TGGAGAGCGG ATCC
Cross-references
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GenBank X56024
UniProt P29064
Protein CAA39501
RefSeq Protein
Broad
PDB
Entry history
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Entry Name AGL31A_CANTS
Previous Entry Names
Last Modification Date 2014-10-12