Applied Microbiology and Biotechnology

, Volume 98, Issue 5, pp 2121–2131 | Cite as

Maltose-forming α-amylase from the hyperthermophilic archaeon Pyrococcus sp. ST04

  • Jong-Hyun Jung
  • Dong-Ho Seo
  • James F. Holden
  • Cheon-Seok ParkEmail author
Biotechnologically relevant enzymes and proteins


The deduced amino acid sequence from a gene of the hyperthermophilic archaeon Pyrococcus sp. ST04 (Py04_0872) contained a conserved glycoside hydrolase family 57 (GH57) motif, but showed <13 % sequence identity with other known Pyrococcus GH57 enzymes, such as 4-α-glucanotransferase (EC, amylopullulanase (EC, and branching enzyme (EC This gene was cloned and expressed in Escherichia coli, and the recombinant product (P yrococcus sp. ST04 maltose-forming α-amylase, PSMA) was a novel 70-kDa maltose-forming α-amylase. PSMA only recognized maltose (G2) units with α-1,4 and α-1,6 linkages in polysaccharides (e.g., starch, amylopectin, and glycogen) and hydrolyzed pullulan very poorly. G2 was the primary end product of hydrolysis. Branched cyclodextrin (CD) was only hydrolyzed along its branched maltooligosaccharides. 6-O-glucosyl-β-cyclodextrin (G1-β-CD) and β-cyclodextrin (β-CD) were resistant to PSMA suggesting that PSMA is an exo-type glucan hydrolase with α-1,4- and α-1,6-glucan hydrolytic activities. The half-saturation value (K m) for the α-1,4 linkage of maltotriose (G3) was 8.4 mM while that of the α-1,6 linkage of 6-O-maltosyl-β-cyclodextrin (G2-β-CD) was 0.3 mM. The k cat values were 381.0 min−1 for G3 and 1,545.0 min−1 for G2-β-CD. The enzyme was inhibited competitively by the reaction product G2, and the K i constant was 0.7 mM. PSMA bridges the gap between amylases that hydrolyze larger maltodextrins and α-glucosidase that feeds G2 into glycolysis by hydrolyzing smaller glucans into G2 units.


Pyrococcus sp. ST04 Hyperthermophile Maltose-forming α-amylase Maltose inhibition 



This work was supported by a National Research Foundation of Korea (NRF) grant funded by the Korean government (MEST; no. 2012–0005289).

Supplementary material

253_2013_5068_MOESM1_ESM.docx (430 kb)
ESM 1 (DOCX 429 kb)


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Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Jong-Hyun Jung
    • 1
  • Dong-Ho Seo
    • 1
  • James F. Holden
    • 2
  • Cheon-Seok Park
    • 1
    • 3
    Email author
  1. 1.Graduate School of Biotechnology and Institute of Life Science and ResourcesKyung Hee UniversityYonginSouth Korea
  2. 2.Department of MicrobiologyUniversity of MassachusettsAmherstUSA
  3. 3.Department of Food Science and Biotechnology and Institute of Life Science and ResourcesKyung Hee UniversityYonginSouth Korea

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