Aggregation dependent enhancement of trehalose-6-phosphate synthase activity in Saccharomyces cerevisiae.
Trehalose-6-phosphate synthase (TPS) is one of the key subunits of the trehalose synthase complex, responsible for synthesis of trehalose in Saccharomyces cerevisiae. Different laboratories have tried to purify TPS, but have been unable to separate it from the complex. During the present study, active TPS has been isolated from the trehalose synthase complex as a free 59 kDa protein. A 15.8 [corrected] fold purification was achieved with over 84% recovery of active TPS. N-terminal sequence confirmed the 59 kDa protein to be TPS. It was revealed to be a highly hydrophobic protein by amino acid analysis data. Activity of TPS was identified to be governed by association-dissociation of protein components. TPS activity of the isolated enzyme was highly unstable due to dissociation of the protein from the complex. Aggregation of active molecules was also seen to enhance as well as stabilize enzyme activity. This aggregation was concentration dependent and activity was seen to be enhanced by increasing the number of active molecules and fell with dilution. The association of the active complex was also found to be governed by ionic interactions.
Chaudhuri
Paramita
P
Biotechnology Division, Indian Institute of Chemical Biology, 4 Raja S. C. Mullick Road, Kolkata 700 032, India.
Basu
Arghya
A
Ghosh
Anil K
AK
D000595:Amino Acid Sequence; D004795:Enzyme Stability; D005964:Glucosyltransferases / Q000737:chemistry* / Q000302:isolation & purification; D057927:Hydrophobic and Hydrophilic Interactions; D008969:Molecular Sequence Data; D009994:Osmolar Concentration; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000254:growth & development; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000302:isolation & purification; D019287:Zinc Sulfate
D029701:Saccharomyces cerevisiae Proteins; D019287:Zinc Sulfate; D005964:Glucosyltransferases; C023180:trehalose-6-phosphate synthase