Two thermophilic and thermostable enzymes, trehalosyl dextrins forming enzyme (TDFE) and trehalose forming enzyme (TFE), able to convert starch and dextrins to alpha,alpha -trehalose were recently purified and characterized from Sulfolobales [I. Di Lernia, A. Morana, A. Ottombrino, S. Fusco, M. Rossi, M. De Rosa, Extremophiles, 2 (1998) 409; T. Nakada, S. Ikegami, H. Chaen, M. Kubota, S. Fukuda, T. Sugimoto, M. Kurimoto, Y. Tsujisaka, Biosci., Biotechnol., Biochem., 60 (1996) 267; T. Nakada, S. Ikegami, H. Chaen, M. Kubota, S. Fukuda, T. Sugimoto, M. Kurimoto, Y. Tsujisaka, Biosci., Biotechnol., Biochem., 60 (1996) 263; M. Kato, Y. Miura, M. Kettoku, K. Shindo, A. Iwamatsu, K. Kobayashi, Biosci., Biotechnol., Biochem., 60 (1996) 921; M. Kato, Y. Miura, M. Kettoku, K. Shindo, A. Iwamatsu, K. Kobayashi, Biosci., Biotechnol., Biochem., 60 (1996) 925]. The first enzyme transforms starch and dextrins to the corresponding trehalosyl derivatives, with an intramolecular transglycosylation process, which converts the glucosidic linkage at the reducing end from alpha -1,4 to alpha -1,1. The second, hydrolyzes the alpha -1,4 Linkage adjacent to the alpha -1,1 bond of trehalosyl dextrins, forming trehalose and lower molecular weight dextrins. Herein, we report the cloning and high level expression of the two enzymes of Sulfolobus solfataricus;strain MT4 in Escherichia coli using pTrc expression vector. The yield of TDFE and TFE obtained in this expression system was of 180 U/1 and of 3630 U/1 of medium, respectively. (C) 2001 Elsevier Science B.V. All rights reserved.
Recombinant thermophilic enzymes for trehalose and trehalosyl dextrins production
de Pascale D;
2001
Abstract
Two thermophilic and thermostable enzymes, trehalosyl dextrins forming enzyme (TDFE) and trehalose forming enzyme (TFE), able to convert starch and dextrins to alpha,alpha -trehalose were recently purified and characterized from Sulfolobales [I. Di Lernia, A. Morana, A. Ottombrino, S. Fusco, M. Rossi, M. De Rosa, Extremophiles, 2 (1998) 409; T. Nakada, S. Ikegami, H. Chaen, M. Kubota, S. Fukuda, T. Sugimoto, M. Kurimoto, Y. Tsujisaka, Biosci., Biotechnol., Biochem., 60 (1996) 267; T. Nakada, S. Ikegami, H. Chaen, M. Kubota, S. Fukuda, T. Sugimoto, M. Kurimoto, Y. Tsujisaka, Biosci., Biotechnol., Biochem., 60 (1996) 263; M. Kato, Y. Miura, M. Kettoku, K. Shindo, A. Iwamatsu, K. Kobayashi, Biosci., Biotechnol., Biochem., 60 (1996) 921; M. Kato, Y. Miura, M. Kettoku, K. Shindo, A. Iwamatsu, K. Kobayashi, Biosci., Biotechnol., Biochem., 60 (1996) 925]. The first enzyme transforms starch and dextrins to the corresponding trehalosyl derivatives, with an intramolecular transglycosylation process, which converts the glucosidic linkage at the reducing end from alpha -1,4 to alpha -1,1. The second, hydrolyzes the alpha -1,4 Linkage adjacent to the alpha -1,1 bond of trehalosyl dextrins, forming trehalose and lower molecular weight dextrins. Herein, we report the cloning and high level expression of the two enzymes of Sulfolobus solfataricus;strain MT4 in Escherichia coli using pTrc expression vector. The yield of TDFE and TFE obtained in this expression system was of 180 U/1 and of 3630 U/1 of medium, respectively. (C) 2001 Elsevier Science B.V. All rights reserved.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
