Crystal structure of a maltogenic amylase provides insights into a catalytic versatility

Jeong Sun Kim, Sun Shin Cha, Hyun Ju Kim, Tae Jip Kim, Nam Chul Ha, Sang Taek Oh, Hyun Soo Cho, Moon Ju Cho, Myo Jeong Kim, Hee Seob Lee, Jung Wan Kim, Kwan Yong Choi, Kwan Hwa Park, Byung Ha Oh

Research output: Contribution to journalArticle

143 Citations (Scopus)

Abstract

Amylases catalyze the hydrolysis of starch material and play central roles in carbohydrate metabolism. Compared with many different amylases that are able to hydrolyze only α-D-(1,4)-glycosidic bonds, maltogenic amylases exhibit catalytic versatility: hydrolysis of α-D-(1,4)-and α-D-(1,6)- glycosidic bonds and transglycosylation of oligosaccharides to C3, C4-, or C6-hydroxyl groups of various acceptor mono-or disaccharides. It has been speculated that the catalytic property of the enzymes is linked to the additional ~130 residues at the N terminus that are absent in other typical α-amylases. The crystal structure of a maltogenic amylase from a Thermus strain was determined at 2.8 Å. The structure, an analytical centrifugation, and a size exclusion column chromatography proved that the enzyme is a dimer in solution. The N-terminal segment of the enzyme folds into a distinct domain and comprises the enzyme active site together with the central (α/β)8 barrel of the adjacent subunit. The active site is a narrow and deep cleft suitable for binding cyclodextrins, which are the preferred substrates to other starch materials. At the bottom of the active site cleft, an extra space, absent in the other typical α-amylases, is present whose size is comparable with that of a disaccharide. The space is most likely to host an acceptor molecule for the transglycosylation and to allow binding of a branched oligosaccharide for hydrolysis of α-D-(1,4)-glycosidic or α-D- (1,6)-glycosidic bond. The (α/β)8 barrel of the enzyme is the preserved scaffold in all the known amylases. The structure represents a novel example of how an enzyme acquires a different substrate profile and a catalytic versatility from a common active site and represents a framework for explaining the catalytic activities of transglycosylation and hydrolysis of α-D-(1,6)-glycosidic bond.

Original languageEnglish
Pages (from-to)26279-26286
Number of pages8
JournalJournal of Biological Chemistry
Volume274
Issue number37
DOIs
Publication statusPublished - 1999 Sep 10

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glucan 1,4-alpha-maltohydrolase
Amylases
Crystal structure
Hydrolysis
Catalytic Domain
Enzymes
Disaccharides
Oligosaccharides
Starch
Thermus
Column chromatography
Size exclusion chromatography
Centrifugation
Carbohydrate Metabolism
Cyclodextrins
Substrates
Scaffolds
Hydroxyl Radical
Dimers
Gel Chromatography

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Kim, J. S., Cha, S. S., Kim, H. J., Kim, T. J., Ha, N. C., Oh, S. T., ... Oh, B. H. (1999). Crystal structure of a maltogenic amylase provides insights into a catalytic versatility. Journal of Biological Chemistry, 274(37), 26279-26286. https://doi.org/10.1074/jbc.274.37.26279
Kim, Jeong Sun ; Cha, Sun Shin ; Kim, Hyun Ju ; Kim, Tae Jip ; Ha, Nam Chul ; Oh, Sang Taek ; Cho, Hyun Soo ; Cho, Moon Ju ; Kim, Myo Jeong ; Lee, Hee Seob ; Kim, Jung Wan ; Choi, Kwan Yong ; Park, Kwan Hwa ; Oh, Byung Ha. / Crystal structure of a maltogenic amylase provides insights into a catalytic versatility. In: Journal of Biological Chemistry. 1999 ; Vol. 274, No. 37. pp. 26279-26286.
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abstract = "Amylases catalyze the hydrolysis of starch material and play central roles in carbohydrate metabolism. Compared with many different amylases that are able to hydrolyze only α-D-(1,4)-glycosidic bonds, maltogenic amylases exhibit catalytic versatility: hydrolysis of α-D-(1,4)-and α-D-(1,6)- glycosidic bonds and transglycosylation of oligosaccharides to C3, C4-, or C6-hydroxyl groups of various acceptor mono-or disaccharides. It has been speculated that the catalytic property of the enzymes is linked to the additional ~130 residues at the N terminus that are absent in other typical α-amylases. The crystal structure of a maltogenic amylase from a Thermus strain was determined at 2.8 {\AA}. The structure, an analytical centrifugation, and a size exclusion column chromatography proved that the enzyme is a dimer in solution. The N-terminal segment of the enzyme folds into a distinct domain and comprises the enzyme active site together with the central (α/β)8 barrel of the adjacent subunit. The active site is a narrow and deep cleft suitable for binding cyclodextrins, which are the preferred substrates to other starch materials. At the bottom of the active site cleft, an extra space, absent in the other typical α-amylases, is present whose size is comparable with that of a disaccharide. The space is most likely to host an acceptor molecule for the transglycosylation and to allow binding of a branched oligosaccharide for hydrolysis of α-D-(1,4)-glycosidic or α-D- (1,6)-glycosidic bond. The (α/β)8 barrel of the enzyme is the preserved scaffold in all the known amylases. The structure represents a novel example of how an enzyme acquires a different substrate profile and a catalytic versatility from a common active site and represents a framework for explaining the catalytic activities of transglycosylation and hydrolysis of α-D-(1,6)-glycosidic bond.",
author = "Kim, {Jeong Sun} and Cha, {Sun Shin} and Kim, {Hyun Ju} and Kim, {Tae Jip} and Ha, {Nam Chul} and Oh, {Sang Taek} and Cho, {Hyun Soo} and Cho, {Moon Ju} and Kim, {Myo Jeong} and Lee, {Hee Seob} and Kim, {Jung Wan} and Choi, {Kwan Yong} and Park, {Kwan Hwa} and Oh, {Byung Ha}",
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Kim, JS, Cha, SS, Kim, HJ, Kim, TJ, Ha, NC, Oh, ST, Cho, HS, Cho, MJ, Kim, MJ, Lee, HS, Kim, JW, Choi, KY, Park, KH & Oh, BH 1999, 'Crystal structure of a maltogenic amylase provides insights into a catalytic versatility', Journal of Biological Chemistry, vol. 274, no. 37, pp. 26279-26286. https://doi.org/10.1074/jbc.274.37.26279

Crystal structure of a maltogenic amylase provides insights into a catalytic versatility. / Kim, Jeong Sun; Cha, Sun Shin; Kim, Hyun Ju; Kim, Tae Jip; Ha, Nam Chul; Oh, Sang Taek; Cho, Hyun Soo; Cho, Moon Ju; Kim, Myo Jeong; Lee, Hee Seob; Kim, Jung Wan; Choi, Kwan Yong; Park, Kwan Hwa; Oh, Byung Ha.

In: Journal of Biological Chemistry, Vol. 274, No. 37, 10.09.1999, p. 26279-26286.

Research output: Contribution to journalArticle

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T1 - Crystal structure of a maltogenic amylase provides insights into a catalytic versatility

AU - Kim, Jeong Sun

AU - Cha, Sun Shin

AU - Kim, Hyun Ju

AU - Kim, Tae Jip

AU - Ha, Nam Chul

AU - Oh, Sang Taek

AU - Cho, Hyun Soo

AU - Cho, Moon Ju

AU - Kim, Myo Jeong

AU - Lee, Hee Seob

AU - Kim, Jung Wan

AU - Choi, Kwan Yong

AU - Park, Kwan Hwa

AU - Oh, Byung Ha

PY - 1999/9/10

Y1 - 1999/9/10

N2 - Amylases catalyze the hydrolysis of starch material and play central roles in carbohydrate metabolism. Compared with many different amylases that are able to hydrolyze only α-D-(1,4)-glycosidic bonds, maltogenic amylases exhibit catalytic versatility: hydrolysis of α-D-(1,4)-and α-D-(1,6)- glycosidic bonds and transglycosylation of oligosaccharides to C3, C4-, or C6-hydroxyl groups of various acceptor mono-or disaccharides. It has been speculated that the catalytic property of the enzymes is linked to the additional ~130 residues at the N terminus that are absent in other typical α-amylases. The crystal structure of a maltogenic amylase from a Thermus strain was determined at 2.8 Å. The structure, an analytical centrifugation, and a size exclusion column chromatography proved that the enzyme is a dimer in solution. The N-terminal segment of the enzyme folds into a distinct domain and comprises the enzyme active site together with the central (α/β)8 barrel of the adjacent subunit. The active site is a narrow and deep cleft suitable for binding cyclodextrins, which are the preferred substrates to other starch materials. At the bottom of the active site cleft, an extra space, absent in the other typical α-amylases, is present whose size is comparable with that of a disaccharide. The space is most likely to host an acceptor molecule for the transglycosylation and to allow binding of a branched oligosaccharide for hydrolysis of α-D-(1,4)-glycosidic or α-D- (1,6)-glycosidic bond. The (α/β)8 barrel of the enzyme is the preserved scaffold in all the known amylases. The structure represents a novel example of how an enzyme acquires a different substrate profile and a catalytic versatility from a common active site and represents a framework for explaining the catalytic activities of transglycosylation and hydrolysis of α-D-(1,6)-glycosidic bond.

AB - Amylases catalyze the hydrolysis of starch material and play central roles in carbohydrate metabolism. Compared with many different amylases that are able to hydrolyze only α-D-(1,4)-glycosidic bonds, maltogenic amylases exhibit catalytic versatility: hydrolysis of α-D-(1,4)-and α-D-(1,6)- glycosidic bonds and transglycosylation of oligosaccharides to C3, C4-, or C6-hydroxyl groups of various acceptor mono-or disaccharides. It has been speculated that the catalytic property of the enzymes is linked to the additional ~130 residues at the N terminus that are absent in other typical α-amylases. The crystal structure of a maltogenic amylase from a Thermus strain was determined at 2.8 Å. The structure, an analytical centrifugation, and a size exclusion column chromatography proved that the enzyme is a dimer in solution. The N-terminal segment of the enzyme folds into a distinct domain and comprises the enzyme active site together with the central (α/β)8 barrel of the adjacent subunit. The active site is a narrow and deep cleft suitable for binding cyclodextrins, which are the preferred substrates to other starch materials. At the bottom of the active site cleft, an extra space, absent in the other typical α-amylases, is present whose size is comparable with that of a disaccharide. The space is most likely to host an acceptor molecule for the transglycosylation and to allow binding of a branched oligosaccharide for hydrolysis of α-D-(1,4)-glycosidic or α-D- (1,6)-glycosidic bond. The (α/β)8 barrel of the enzyme is the preserved scaffold in all the known amylases. The structure represents a novel example of how an enzyme acquires a different substrate profile and a catalytic versatility from a common active site and represents a framework for explaining the catalytic activities of transglycosylation and hydrolysis of α-D-(1,6)-glycosidic bond.

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