A NOVEL LIMONOID UDP-GLUCOSYLTRANSFERASE AND METHOD FOR PREPARING THE SAME

摘要

A novel limonoid UDP(uracil diphosphate)-glucosyltransferase is provided to remove bitter taste of limonoid while maintaining its physiological activity by converting limonoid into limonin glucoside, so that the enzyme is useful for preparing processed tangerine products having no bitter taste. A novel limonoid UDP-glucosyltransferase has the amino acid sequence of SEQ ID NO:1. A gene encoding the limonoid UDP-glucosyltransferase has the nucleotide sequence of SEQ ID NO:2. A method for producing the limonoid UDP-glucosyltransferase comprises the steps of: preparing a recombinant vector containing the limonoid UDP-glucosyltransferase gene; transforming a host cell selected from insect cell, bacteria, yeast and fungi with the recombinant vector to obtain a transformed microorganism; and culturing the transformed microorganism. A method for highly expressing the limonoid UDP-glucosyltransferase gene comprises the steps of: (a) transducing Escherichia coli with the recombinant vector containing the limonoid UDP-glucosyltransferase gene to produce a recombinant vector containing the limonoid UDP-glucosyltransferase gene; (b) transfecting the cloning vector to an insect cell such as Spodoptera frugiperda(Sf9) by using lipofectamine; and (c) culturing the transfected insect cell, and isolating and purifying the limonoid UDP-glucosyltransferase.