Purification and characterization of a novel cold adapted fungal glucoamylase

• Published in: Microbial Cell Factories
• Main Authors: Carrasco, Mario, Alcaíno Gorman, Jennifer, Cifuentes Guzmán, Víctor, Baeza Cancino, Marcelo
• Format: Article in Journal/Newspaper
• Language: English
• Published: BioMed Central 2017
• Subjects: Antarctic fungi; Cold-adapted amylase; Fungal amylase; Tetracladium sp; Antarctic; Antarc*
• Online Access: https://doi.org/10.1186/s12934-017-0693-x; https://repositorio.uchile.cl/handle/2250/168936

Background: Amylases are used in various industrial processes and a key requirement for the efficiency of these processes is the use of enzymes with high catalytic activity at ambient temperature. Unfortunately, most amylases isolated from bacteria and filamentous fungi have optimal activity above 45 °C and low pH. For example, the most commonly used industrial glucoamylases, a type of amylase that degrades starch to glucose, are produced by Aspergillus strains displaying optimal activities at 45–60 °C. Thus, isolating new amylases with optimal activity at ambient temperature is essential for improving industrial processes. In this report, a glucoamylase secreted by the cold-adapted yeast Tetracladium sp. was isolated and biochemically characterized. Results: The effects of physicochemical parameters on enzyme activity were analyzed, and pH and temperature were found to be key factors modulating the glucoamylase activity. The optimal conditions for enzyme activity were 30 °C and pH 6.0, and the Km and kcat using soluble starch as substrate were 4.5 g/L and 45 min−1, respectively. Possible amylase or glucoamylase encoding genes were identified, and their transcript levels using glucose or soluble starch as the sole carbon source were analyzed. Transcription levels were highest in medium supplemented with soluble starch for the potential glucoamylase encoding gene. Comparison of the structural model of the identified Tetracladium sp. glucoamylase with the solved structure of the Hypocrea jecorina glucoamylase revealed unique structural features that may explain the thermal lability of the glucoamylase from Tetracladium sp.