Abstract
Saccharomycopsis (Syn. Endomycopsis) bubodii 2066 is an isolate from bubod, a starter used in making rice wine in northern Philippines. We have shown that the yeast has amylolytic activity on raw sago starch. In our attempt to identify the putative raw starch-digesting amylase in S. bubodii, we determined the cDNA sequence of a glucoamylase gene. One primer pair that was designed based on a glucoamylase of Saccharomycopsis fibuligera HUT7212 (GLU1, NCBI Accession Number L25641.1) produced a sequence of 1234 base pairs. To obtain a wider coverage, a primer walking strategy was carried out using four primer pairs designed based on GLU1 gene. The generated sequence of 1535 base pairs shows 98.7 to 100% homology when aligned with glucoamylase genes from four strains of S. fibuligera suggesting that this glucoamylase is highly conserved between the Saccharomycopsis species. This work further reports a gene sequence of glucoamylase derived from Philippine-isolated yeast. The sequence is deposited in GenBank and assigned the accession number KP068007.1. The gene may be heterologously expressed in Saccharomyces cerevisiae for possible utilization in the direct conversion of raw sago starch to bioethanol.
Highlights
This study showed that Saccharomycopsis bubodii 2066 is a potential source of raw-starch digesting amylase (RSDA)
Another yeast strain identified as Saccharomycopsis fibuligera 2074 which is obtained from Bubod, displayed the same preference for raw sago starch (Bullo, 2009)
Endomycopsis) bubodii 2066 was shown to exhibit amylolytic activity on raw sago starch indicating the yeast as potential source of rawstarch digesting amylase (RSDA)
Summary
Saccharomycopsis (Syn. Endomycopsis) bubodii 2066 was isolated from bubod, a starter culture used in making the rice wine locally called tapuy in Tublay, Benguet, Philippines (PNMCC Directory of Strains, 2012). Endomycopsis) bubodii 2066 was isolated from bubod, a starter culture used in making the rice wine locally called tapuy in Tublay, Benguet, Philippines (PNMCC Directory of Strains, 2012) This strain of filamentous yeast was studied taxonomically and was identified as a new species of the genus Endomycopsis (Saccharomycopsis) by Sakai and Caldo (Sakai and Caldo, 1985b). The genetic diversity of yeast isolates from Philippine rice wine was analyzed using modified Random Amplification of Polymorphic DNA (RAPD) with 20-mer Seoulin Research Institute Life Science (SRILS) uniprimers 1, 6 and 9. Dendrogram analysis by NTSYS based on banding patterns generated through these uniprimers showed S. bubodii 2066 as divergent yeast with only around 43% similarity with the Saccharomycopsis fibuligera strains (Lim et al, 2006).
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