ABSTRACT
Torulaspora delbrueckii is a yeast species receiving increasing attention from the biotechnology industry, with particular relevance in the wine, beer and baking sectors. However, little is known about its sugar transporters and sugar transport capacity, frequently a rate-limiting step of sugar metabolism and efficient fermentation. Actually, only one glucose transporter, Lgt1, has been characterized so far. Here we report the identification and characterization of a second glucose transporter gene, IGT1, located in a cluster, upstream of LGT1 and downstream of two other putative hexose transporters. Functional characterization of IGT1 in a Saccharomyces cerevisiae hxt-null strain revealed that it encodes a transporter able to mediate uptake of glucose, fructose and mannose and established that its affinity, as measured by Km, could be modulated by glucose concentration in the medium. In fact, IGT1-transformed S. cerevisiae hxt-null cells, grown in 0.1% glucose displayed biphasic glucose uptake kinetics with an intermediate- (Km = 6.5 ± 2.0 mM) and a high-affinity (Km = 0.10 ± 0.01 mM) component, whereas cells grown in 2% glucose displayed monophasic kinetics with an intermediate-affinity (Km of 11.5 ± 1.5 mM). This work contributes to a better characterization of glucose transport in T. delbrueckii, with relevant implications for its exploitation in the food industry.
Subject(s)
Carbohydrate Metabolism , Glucose/metabolism , Monosaccharide Transport Proteins/genetics , Torulaspora/genetics , Torulaspora/metabolism , Fermentation , Fructose/metabolism , Kinetics , Mannose/metabolism , Monosaccharide Transport Proteins/metabolism , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/metabolismABSTRACT
SCOPE: Previous results suggested that the effects of Salvia fruticosa tea (SFT) drinking on glucose regulation might be at the intestinal level. Here we aim to characterize the effects of SFT treatment and of its main phenolic constituent--rosmarinic acid (RA)--on the levels and localization of the intestinal Na+/glucose cotransporter-1 (SGLT1), the facilitative glucose transporter 2 and glucagon-like peptide-1 (GLP-1). METHODS AND RESULTS: Two models of SGLT1 induction in rats were used: through diabetes induction with streptozotocin (STZ) and through dietary carbohydrate manipulation. Drinking water was replaced with SFT or RA and blood parameters, liver glycogen and the levels of different proteins in enterocytes quantified. Two weeks of SFT treatment stabilized fasting blood glucose levels in STZ-diabetic animals. The increase in SGLT1 localized to the enterocyte brush-border membrane (BBM) induced by STZ treatment was significantly abrogated by treatment with SFT, without significant changes in total cellular transporter protein levels. No effects were observed on glucose transporter 2, Na(+) /K(+) -ATPase or glucagon-like peptide-1 levels by SFT. Additionally, SFT and RA for 4 days significantly inhibited the carbohydrate-induced adaptive increase of SGLT1 in BBM. CONCLUSION: SFT and RA modulate the trafficking of SGLT1 to the BBM and may contribute to the control of plasma glucose.
Subject(s)
Blood Glucose/analysis , Cinnamates/pharmacology , Depsides/pharmacology , Plant Preparations/pharmacology , Salvia officinalis/chemistry , Sodium-Glucose Transporter 1/analysis , Animals , Diabetes Mellitus, Experimental , Enterocytes/metabolism , Glucagon-Like Peptide 1/metabolism , Glucose Transporter Type 2/metabolism , Hypoglycemic Agents/pharmacology , Intestinal Mucosa/metabolism , Jejunum/metabolism , Male , Microvilli/metabolism , Rats , Rats, Wistar , Sodium-Glucose Transporter 1/metabolism , Rosmarinic AcidABSTRACT
PCR-based disruption cassettes are one of the most commonly used strategies for gene targeting in Saccharomyces cerevisiae. The efficiencies of gene disruption using this conventional method are highly variable among species, and often quite low with nonconventional yeasts. Here we describe an improved strategy to obtain deletion mutants in baker's yeast Torulaspora delbrueckii, one of the most abundant non-Saccharomyces species, present in home-made corn and rye bread dough.
Subject(s)
Gene Knockout Techniques/methods , Torulaspora/genetics , Gene Deletion , Secale/microbiology , Zea mays/microbiologyABSTRACT
The freeze stress responses to prolonged storage at -20 degrees C in Torulaspora delbrueckii PYCC5323 were investigated. In this yeast, no loss of cell viability was observed for at least 120 days during freezing at -20 degrees C, whereas a loss of 80% was observed in a commercial baker's yeast after 15 days. In the former strain, freeze resistance was dependent on an adaptation process. The primary cell target of freeze stress was the plasma membrane, preservation of its integrity being related with a lower increase of lipid peroxidation and with a higher resistance to H(2)O(2), but not with the intracellular trehalose concentration.
