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Production of a thermophilic maltooligosyl-trehalose synthase in <Emphasis Type="Italic">Lactococcus lactis</Emphasis>
Authors:Donatella Cimini  Mario De Rosa  Andrea Panariello  Veronica Morelli  Chiara Schiraldi
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.
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.
Keywords:Intramolecular transglycosylation  Nisin induction  Recombinant Lactococcus lactis            Thermozymes  Trehalosyl-dextrins
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