Recombinant thermophilic enzymes for trehalose and trehalosyl dextrins production |
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Authors: | Donatella de Pascale, Maria Paola Sasso, Isabella Di Lernia, Antonella Di Lazzaro, Adriana Furia, Maria Cartenì Farina, Mos Rossi,M. De Rosa |
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Affiliation: | a Dipartimento di Chimica Organica e Biologica, Università di Napoli, Via Mezzocannone 16, Naples 80134, Italy b Istituto di Farmacologia e Tossicologia, Facoltà di Medicina-CRISCEB, Seconda Università di Napoli, Via Costantinopoli 16, Naples 80134, Italy c Istituto di Biochimica delle Proteine ed Enzimologia C.N.R.-CRIB, Via Marconi 10, Naples 80127, Italy |
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Abstract: | Two thermophilic and thermostable enzymes, trehalosyl dextrins forming enzyme (TDFE) and trehalose forming enzyme (TFE), able to convert starch and dextrins to ,-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 -1,4 to -1,1. The second, hydrolyzes the -1,4 linkage adjacent to the -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/l and of 3630 U/l of medium, respectively. |
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Keywords: | Extremophiles Starch Trehalose Trehalose producing enzymes E. coli |
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