The Saccharomyces cerevisiae enolase‐related regions encode proteins that are active enolases

Abstract

In addition to two genes (ENO1 and ENO2) known to code for enolase (EC4.2.1.11), the Saccharomyces cerevisiae genome contains three enolase-related regions (ERR1, ERR2 and ERR3) which could potentially encode proteins with enolase function. Here, we show that products of these genes (Err2p and Err3p) have secondary and quaternary structures similar to those of yeast enolase (Eno1p). In addition, Err2p and Err3p can convert 2-phosphoglycerate to phosphoenolpyruvate, with kinetic parameters similar to those of Eno1p, suggesting that these proteins could function as enolases in vivo. To address this possibility, we overexpressed the ERR2 and ERR3 genes individually in a double-null yeast strain lacking ENO1 and ENO2, and showed that either ERR2 or ERR3 could complement the growth defect in this strain when cells are grown in medium with glucose as the carbon source. Taken together, these data suggest that the ERR genes in Saccharomyces cerevisiae encode a protein that could function in glycolysis as enolase. The presence of these enolase-related regions in Saccharomyces cerevisiae and their absence in other related yeasts suggests that these genes may play some unique role in Saccharomyces cerevisiae. Further experiments will be required to determine whether these functions are related to glycolysis or other cellular processes.

Immunization with recombinant enolase ofSporothrixspp (rSsEno) confers effective protection against sporotrichosis in mice

“…Our results showed that rSsEno was produced successfully with the expected molecular weight of 47 kDa and with the secondary structure of a type α-helix at 46% and a β-sheet at 12%. These values are close to values of 43% for the α-helix and 15% for the β-sheet from Saccharomyces cerevisiae enolase 35 , suggesting that both enolases have similar structures. The analyses of the oligomeric state of rSsEno by gel filtration revealed an unexpected result.…”

Unveiling the potential of novel yeast protein extracts in white wines clarification and stabilization

“…In fact, yeast plays a primary role in the winemaking process and thus, after fermentation, its autolysis naturally leads to a release of diverse cell compounds as enzymes, mannoproteins, fatty acids, nucleotides, peptides, and free amino acids (Zhang et al, 2011 ). Nevertheless, Saccharomyces cerevisiae extracts are known to contain some particular proteins that are potentially involved in allergic responses (Lindberg et al, 1992 ; Kortekangas-Savolainen et al, 1993 ; Nermes et al, 1995 ; Savolainen et al, 1998 ; Nittner-Marszalska et al, 2001 ), namely the glycolytic enzymes, Enolase 1 (ENO1), and Enolase 2 (ENO2) that are involved in carbohydrate metabolism and therefore, intrinsic to the wine fermentation process (Varela et al, 2005 ; Kornblatt et al, 2013 ), contrary to all the exogenous products that are commonly applied and already described to raise both health and environmental problems.…”

Identification of Thermal Conduits That Link the Protein–Water Interface to the Active Site Loop and Catalytic Base in Enolase

“…The primary reaction catalyzed is the abstraction of an α-proton from a carboxylate substrate, with the incipient anionic intermediate undergoing stabilization by a divalent metal cation (usually Mg 2+ ) . As the defining member of its superfamily, enolase is particularly well-studied, with research documented in the 1940s and continuing to the present time. − The enzyme is part of the glycolytic pathway, responsible for converting 2-phospho- d -glycerate (2-PGA) to phosphoenol pyruvate (PEP), and is found across all domains of life. The reaction catalyzed by enolase is reversible, and an X-ray structure has been solved for enolase in the presence of an equilibrium mixture of substrate and product …”

“…Kinetic parameters k cat , kinetic isotope effect (KIE), and K M are calculated from experiments at 25 °C and are an average of at least three experiments. b E a values are calculated from k cat at least five different temperatures(10,20,25,30, and 40 °C) and are the average of at least three experiments (Table…”