Production of a thermophilic maltooligosyl-trehalose synthase in <Emphasis Type="Italic">Lactococcus lactis</Emphasis> |
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Authors: | Donatella Cimini Mario De Rosa Andrea Panariello Veronica Morelli Chiara Schiraldi |
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Institution: | Department of Experimental Medicine, Section of Biotechnology and Molecular Biology, Faculty of Medicine and Surgery, Second University of Naples, via De Crecchio 7, 80138, Naples, Italy. |
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Abstract: | The thermoacidophilic archaeon Sulfolobus solfataricus MT4 encodes a maltooligosyltrehalose synthase (MTS), that catalyzes an intramolecular transglycosylation process converting the glycosidic linkages at the reducing end of dextrins from alpha-1,4 into alpha-1,1. In this research the gene encoding MTS was cloned and expressed in Lactococcus lactis NZ9000 using the so-called NICE system. Growth conditions of the recombinant strain were optimized in flask experiments in relation to enzyme production. Batch experiments in 2 L-fermenters were performed on the best identified semidefined medium and 256 U L(-1) of recombinant MTS were produced. Purified recombinant MTS shows its optimal activity at 70 degrees C and pH 5.5, prefers maltoheptaose and maltohexaose as substrates, and demonstrates minimal side hydrolytic activity. |
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Keywords: | Intramolecular transglycosylation Nisin induction Recombinant Lactococcus lactis Thermozymes Trehalosyl-dextrins |
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