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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 3′half 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  YlTEF1 promoter or with a disruption of YALI0D15598 encoding a putative trehalase. mRNA levels of  YlTPS1  were low and did not respond to thermal stresses, but that of  YlTPS2  (YALI0D14476) and  YlTPS3  (YALI0E31086) increased 4 and 6 times, repectively, by heat treatment. Disruption of  YlTPS1  drastically slowed growth at 35°C. Homozygous  Yltps1  diploids showed a decreased sporulation frequency that was ascribed to the low level of YALI0D20966 mRNA an homolog of the  S. cerevisiae MCK1  which encodes a protein kinase that activates early meiotic gene expression.

Disruption of Yarrowia lipolytica TPS1 Gene Encoding Trehalose-6-P Synthase Does Not Affect Growth in Glucose but Impairs Growth at High Temperature