disruption of yarrowia lipolytica tps1 gene encoding trehalose-6-p synthase does not affect growth in glucose but impairs growth at high temperature中断yarrowia lipolytica tps1基因编码trehalose-6-p合酶葡萄糖不影响增长但损害增长在高温下.pdfVIP

Disruption of Yarrowia lipolytica TPS1 Gene Encoding Trehalose-6-P Synthase Does Not Affect Growth in Glucose but Impairs Growth at High Temperature 1 1 2 Carmen-Lisset Flores *, Carlos Gancedo , Thomas Petit ´ ´ ´ 1 Department of Metabolism and Cell Signalling, Instituto de Investigaciones Biomedicas ‘‘Alberto Sols’’ CSIC-UAM, Madrid, Spain, 2 Universite de La Reunion, Laboratoire ´ ´ de Chimie des Substances Naturelles et des Sciences des Aliments, Institut Universitaire de Technologie, Departement Genie Biologique, Saint Pierre, France Abstract We have cloned the Yarrowia lipolytica TPS1 gene encoding trehalose-6-P synthase by complementation of the lack of growth in glucose of a Saccharomyces cerevisiae tps1 mutant. Disruption of YlTPS1 could only be achieved with a cassette placed in the 39half of its coding region due to the overlap of its sequence with the promoter of the essential gene YlTFC1. The Yltps1 mutant grew in glucose although the Y. lipolytica hexokinase is extremely sensitive to inhibition by trehalose-6-P. The presence of a glucokinase, insensitive to trehalose-6-P, that constitutes about 80% of the glucose phosphorylating capacity during growth in glucose may account for the growth phenotype. Trehalose content was below 1 nmol/mg dry weight in Y. lipolytica, but it increased in strains expressing YlTPS1 under the control of the YlTEF1promoter or with a disruption of YALI0D15598 encoding a putative trehalase. mRNA levels of YlTPS1 were low and did not respond to thermal stress