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Determination of the cDNA Sequence and In Silico Functional Analysis of a Glucoamylase Gene From Saccharomycopsis fibuligera 2074

Dan Exerlin E. Bonete, Joel Hassan G. Tolentino, and Annabelle U. Novero*

College of Science and Mathematics, University of the Philippines Mindanao,
Mintal, Tugbok District, Davao City 8022 Philippines

*Corresponding author: This email address is being protected from spambots. You need JavaScript enabled to view it.

 

ABSTRACT

Saccharomycopsis fibuligera 2074 is a yeast strain used in producing tapuy, a traditional Philippine wine mix. A glucoamylase gene from this yeast was isolated and characterized in this study. Using primers designed via the primer walking method, the synthesized Sf 2074 cDNA (GenBank: KP068008.1) was found to contain 1531 bp and was homologous to glucoamylases deposited in the databases. Using bioinformatic tools, the predicted protein was found to possess 512 amino acids and a molecular weight of 56715.92. The conserved amino acid sequence Ala-Tyr-Thr-Gly similar to other amylases was located. The glucoamylase belongs to a superfamily of Glycoside Hydrolase 15 (GH 15), which are six-hairpin glycosidases with alpha/alpha toroid fold. This is the first report of a glucoamylase gene from S. fibuligera in the Philippines. Bioethanol cost of production could be markedly reduced if this amylolytic gene can be cloned in the brewer’s yeast Saccharomyces cerevisieae.

Keywords: glucoamylase, in silico characterization,  primer walking, protein structure, Saccharomycopsis fibuligera, yeast

 

INTRODUCTION

Ethanol is the most widely used liquid biofuel. The demand for ethanol is expected to rise to over 125 billion liters in 2017 (FAO 2008). It is fermented from sugars, starches, or from cellulosic biomass. Production of ethanol from starch is one way to reduce consumption of crude oil as well as environmental pollution. In view of continuously rising petroleum costs and dependence upon fossil fuel resources, considerable attention has been focused on alternative energy resources. Production of ethanol from biomass is one way to reduce both the consumption of crude oil and environmental pollution (DiPardo 2000; Bothast & Schlicher 2005; Dufey 2006; Schafer et al. 2007). . . . . read more

 

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