Glycerol Production in a Triose Phosphate Isomerase Deficient Mutant of Saccharomyces cerevisiae

Abstract

Interesting challenges from metabolically engineered Saccharomyces cerevisiae cells arise from the opportunity to obtain yeast strains useful for the production of chemicals. In this paper, we show that engineered yeast cells deficient in the triose phosphate isomerase activity are able to produce glycerol without the use of steering agents. High yields of conversion of glucose into glycerol (80-90% of the theoretical yield) and productivity (1.5 g L-1 h-1) have been obtained by a bioconversion process carried out in a poor and clean medium. We obtained indications that the growth phase at which the biomass was collected affect the process. The best results were obtained using cells collected at the end of exponential phase of growth. In perspective, the strategies and the information about the physiology of the cells described here could be useful for the developing of new biotechnological processes for glycerol production, outflanking the problems related to the use of high level of steering agents.

Isolation and sequence analysis of the gene encoding triose phosphate isomerase from Zygosaccharomyces bailii

“…In S. cerevisiae the lack of TIM activity causes an accumulation of DHAP, which leads to a redirection of the NADH reoxidation (Compagno et al, 1996). In such a mutant, a high synthesis of glycerol was observed (Compagno et al, 1996). In Kluyveromyces lactis, a similar effect has been recently reported (Compagno et al, 1999).…”

“…Cloning of the Z. bailii gene complementing a S. cerevisiae tpi1 mutant A S. cerevisiae strain lacking triose phosphate isomerase activity is unable to grow on glucose media (Compagno et al, 1996). We used this defective phenotype to search for the Z. bailii gene coding for TIM by functional complementation.…”

“…bailii ISA 1307, originally isolated as a spoilage agent from a continuous production plant of sparkling wine (Wium et al, 1990), was obtained from the Culture Collection of the Instituto Superior de Agronomia (Lisbon, Portugal). S. cerevisiae W303DTPI1 (MATa, tpi1::kan r , ade2-1, can1-100, ura3-1, leu2-3,112, trp1-1, his3-11,15; Compagno et al, 1996) displays the TIM-deficient phenotype and was used for complementation assays. Cells of S. cerevisiae W303DTPI1 were grown in rich media (2% w/v peptone, 1% w/v yeast extract) or minimal media (6.7 g/l Yeast Nitrogen Base, adenine 100 mg/l, uracil 50 mg/l, leucine 50 mg/l, tryptophan 50 mg/l, histidine 50 mg/l) containing both glucose (0.1%, w/v) and ethanol (1%, v/v) as carbon and energy sources.…”

“…It catalyses the interconversion between dihydroxyacetone phosphate (DHAP) and glyceraldehyde-3-phosphate (GAP), which fall into important metabolic branch points between glycolysis, gluconeogenesis, the pentose pathway, glycerol production/utilization and the methylglyoxal pathway (Gancedo and Serrano, 1989;Cooper, 1984). In S. cerevisiae the lack of TIM activity causes an accumulation of DHAP, which leads to a redirection of the NADH reoxidation (Compagno et al, 1996). In such a mutant, a high synthesis of glycerol was observed (Compagno et al, 1996).…”

Metabolic Control Analysis and its Applications

“…It is used to synthetize products like lubricants and 8 cosmetics [67] and it is also used as an antifreeze in chemical industry [68].…”