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Applied and Environmental Microbiology, July 2004, p. 3933-3940, Vol. 70, No. 7
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.7.3933-3940.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Properties of a Novel Thermostable Glucoamylase from the Hyperthermophilic Archaeon Sulfolobus solfataricus in Relation to Starch Processing

Mi-Sun Kim,1,{dagger} Jong-Tae Park,1,{dagger} Young-Wan Kim,1 Hee-Seob Lee,1 Rose Nyawira,1 Hyoun-Seung Shin,2 Cheon-Seok Park,3 Sang-Ho Yoo,4 Yong-Ro Kim,1 Tae-Wha Moon,1 and Kwan-Hwa Park1*

National Research Laboratory for Functional Food Carbohydrate and Center for Agricultural Biomaterials, Seoul National University, Sillim-dong, Kwanak-gu, Seoul 151-742,1 Bolak Co., Ltd., Hwaseong 445-933,2 Department of Food Science and Technology, Kyunghee University, Youngin 449-701,3 Department of Food Science and Technology, Sejong University, Seoul 143-747, Korea4

Received 17 December 2003/ Accepted 1 April 2004

A gene (ssg) encoding a putative glucoamylase in a hyperthermophilic archaeon, Sulfolobus solfataricus, was cloned and expressed in Escherichia coli, and the properties of the recombinant protein were examined in relation to the glucose production process. The recombinant glucoamylase was extremely thermostable, with an optimal temperature at 90°C. The enzyme was most active in the pH range from 5.5 to 6.0. The enzyme liberated ß-D-glucose from the substrate maltotriose, and the substrate preference for maltotriose distinguished this enzyme from fungal glucoamylases. Gel permeation chromatography and sedimentation equilibrium analytical ultracentrifugation analysis revealed that the enzyme exists as a tetramer. The reverse reaction of the glucoamylase from S. solfataricus produced significantly less isomaltose than did that of industrial fungal glucoamylase. The glucoamylase from S. solfataricus has excellent potential for improving industrial starch processing by eliminating the need to adjust both pH and temperature.


* Corresponding author. Mailing address: National Research Laboratory for Functional Food Carbohydrate and Center for Agricultural Biomaterials, Seoul National University, Sillim-dong, Kwanak-gu, Seoul 151-742, Korea. Phone: 82-2-8804852. Fax: 82-2-8735095. E-mail: parkkh{at}plaza.snu.ac.kr.

{dagger} M.-S.K. and J.-T.P. contributed equally to this work.


Applied and Environmental Microbiology, July 2004, p. 3933-3940, Vol. 70, No. 7
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.7.3933-3940.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.