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Applied and Environmental Microbiology, August 2000, p. 3350-3356, Vol. 66, No. 8
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Comparative Characterization of Complete and Truncated Forms of Lactobacillus amylovorus alpha -Amylase and Role of the C-Terminal Direct Repeats in Raw-Starch Binding

R. Rodriguez Sanoja,1 J. Morlon-Guyot,1,* J. Jore,2 J. Pintado,1 N. Juge,3 and J. P. Guyot1

Laboratoire de Biotechnologie Microbienne Tropicale, Institut de Recherche pour le Développement, 34032 Montpellier cedex 1, France1; TNO VOEDING, Dept. AMGT, 3700 AJ Zeist, The Netherlands2; and Institute of Food Research, Norwich Research Park, Obrey, Norwich NR4 7UA, United Kingdom3

Received 6 March 2000/Accepted 3 May 2000

Two constructs derived from the alpha -amylase gene (amyA) of Lactobacillus amylovorus were expressed in Lactobacillus plantarum, and their expression products were purified, characterized, and compared. These products correspond to the complete (AmyA) and truncated (AmyADelta ) forms of alpha -amylase; AmyADelta lacks the 66-kDa carboxyl-terminal direct-repeating-unit region. AmyA and AmyADelta exhibit similar amylase activities towards a range of soluble substrates (amylose, amylopectin and alpha -cyclodextrin, and soluble starch). The specific activities of the enzymes towards soluble starch are similar, but the KM and Vmax values of AmyADelta were slightly higher than those of AmyA, whereas the thermal stability of AmyADelta was lower than that of AmyA. In contrast to AmyA, AmyADelta is unable to bind to beta -cyclodextrin and is only weakly active towards glycogen. More striking is the fact that AmyADelta cannot bind or hydrolyze raw starch, demonstrating that the carboxyl-terminal repeating-unit domain of AmyA is required for raw-starch binding activity.

* Corresponding author. Mailing address: I.R.D. (ORSTOM), Laboratoire de Biotechnologie Microbienne Tropicale (LBMT), BP 5045, 34032 Montpellier cedex 1, France. Phone: 33 4 67 41 62 78. Fax: 33 4 67 54 78 00. E-mail: Juliette.Morlon-Guyot{at}mpl.ird.fr.

Applied and Environmental Microbiology, August 2000, p. 3350-3356, Vol. 66, No. 8
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Guillen, D., Santiago, M., Linares, L., Perez, R., Morlon, J., Ruiz, B., Sanchez, S., Rodriguez-Sanoja, R. (2007). Alpha-Amylase Starch Binding Domains: Cooperative Effects of Binding to Starch Granules of Multiple Tandemly Arranged Domains. Appl. Environ. Microbiol. 73: 3833-3837 [Abstract] [Full Text]  
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