Purification and characterization of extracellular α‐amylase and glucoamylase from the yeast Candida antarctica CBS 6678

Abstract

An a-amylase and a glucoamylase were purified to homogeneity from the culture fluid of /3-cyclodextringrown Candida antarctica CBS 6678 by protamine sulfate treatment, ammonium sulfate precipitation, gel filtration (Sephadex G-75 sf, Ultrogel AcA 54), DEAE-Sephacel chromatography, hydroxyapatite chromatography and affinity chomatography on acarbose -AH-Sepharose 4B.Both enzymes were monomeric glycoproteins with fairly different amino acid compositions. Their apparent relative molecular mass, sedimentation coefficient (s&, , ), isoelectric point, absorption coefficient (280 nm), pH and temperature optima were estimated as 48500, 4.7 S, 10.1, 1.74 cm2 mg-', 4.2 and 57"C, respectively, for glucoamylase and as 50000, 4.9 S, 10.3, 1.53 cm2 mg-l, 4.2 and 62"C, respectively, for a-amylase.Kinetic analyses indicated that both enzymes preferentially hydrolyzed high-molecular-mass substrates, including some raw starches. a-Amylase was active on cyclodextrins, whereas debranching activity was demonstrated for glucoamylase. Trestatins were potent inhibitors of both a-amylase (Ki < 1 pM) and glucoamylase (Ki < 0

Role of maltase in the utilization of sucrose by Candida albicans

“…However, ,J-fructofuranosidase activity was not detected in response to growth in sucrose, in contrast with control experiments using S. cerevisiae A.T.C.C. 26108 (0.19 unit/mg of protein; level of detection < 0.002 unit/mg) as was reported previously (DeMot and Verachtert, 1987).…”

“…This pattern of substrate hydrolysis is similar to most preparations of isoenzymes of maltase from Saccharomyces (Needleman et al, 1978;Siro and Lovrgren, 1978;Krakenaite and Glemzha, 1983;Tabata et al, 1984) except for one preparation in which the two enzymes differed by their ability in turn to hydrolyse maltose or a-methyl D-glucopyranoside (Khan and Eaton, 1967). In addition, the lack of hydrolysis of the a(l -.6) linkage of isomaltose distinguishes this enzyme from a glucoamylase purified from a diastotic strain of S. cerevisiae by Kleinman et al (1988), and the lack of apparent secretion of this enzyme into the surrounding fluid contrasts this enzyme with the glucoamylase from Candida tropicalis (Sawai and Hehre, 1962), Candida antarctica (DeMot and Verachtert, 1987) and Candida tsukubaensis (DeMot et al, 1985).…”

Purification and characterization of a novel α-glucosidase from an Antarctic yeast Dioszegia fristingensis isolate

“…Most of other microbial α-amylases and glucoamylases exhibit their best activity at 40°C, and have lower optimal pH (average 4.5) for optimal activity [20]. The cold-active α-amylase produced by Antarctic bacterium Pseudoalteromonas haloplanktis is best active at 25°C and pH 7.0 [43], whereas the one from the yeast Candida antarctica (now Moesziomyces antarcticus) displayed optimal activity at 57°C and pH 4.2 [30].…”

“…cold-adapted yeasts) represent a promising source for cold-active amylolytic enzymes since they, using the available carbon sources, secrete hydrolytic enzymes, which have potential for biotechnological and industrial applications [26][27][28][29]. An α-amylase and a glucoamylase were described from Candida antarctica (now Moesziomyces antarcticus) CBS 6678 [30], both enzymes being monomeric glycoproteins that preferentially hydrolyzed high-molecular-mass substrates. The optimal pH and temperature for activity of these glucoamylase and α-amylase were 4.2 and 57°C, and 4.2 and 62°C, respectively [30].…”

“…An α-amylase and a glucoamylase were described from Candida antarctica (now Moesziomyces antarcticus) CBS 6678 [30], both enzymes being monomeric glycoproteins that preferentially hydrolyzed high-molecular-mass substrates. The optimal pH and temperature for activity of these glucoamylase and α-amylase were 4.2 and 57°C, and 4.2 and 62°C, respectively [30]. An amylase produced by the yeast Clavispora lusitaniae isolated from vegetables of a market of Bhopal (India) had higher activity at pH 11 and 42°C, maintaining a 42% of its activity at 4°C and showed no dependency on Ca 2+ for activity and stability.…”