Isolation and expression in Saccharomyces cerevisiae of a gene encoding an α‐amylase from Schwanniomyces castellii

Abstract

A gene (SWA1) encoding an α‐amylase activity from Schwanniomyces castellii has been cloned and expressed, via yeast cloning vector YEp13, in Saccharomyces cerevisiae. By using a riboprobe which is internal to the SWA1 gene, a 1.55 kb transcript was detected in the poly(A)+ RNA from both Sw. castellii and a S. cerevisiae clone harboring the SWA1 gene. This transcript should, therefore, correspond to the SWA1 gene. In addition, the DNA strand determining the α‐amylase activity has been defined. Transcription of the SWA1 gene appears to be highly regulated in Sw. castellii, whereas it is constitutive in the S. cerevisiae harboring this gene.

Structural and Kinetic Analysis of Schwanniomyces occidentalis Invertase Reveals a New Oligomerization Pattern and the Role of Its Supplementary Domain in Substrate Binding

“…The nonconventional yeast Schwanniomyces occidentalis (also Debaryomyces occidentalis) has been regarded for years as a biotechnologically interesting organism because of its ability to grow in a broad range of inexpensive carbon sources, such as starch and inulin, using a number of activities (12)(13)(14)(15), as well as for its efficient extracellular secretion of high molecular mass proteins (16,17) and its low attached glycosylation (18). An extracellular invertase/fructofuranosidase activity has also been reported in this yeast when lactose was used as the carbon source (14).…”

Construction of an efficient amylolytic industrial yeast strain containing DNA exclusively derived from yeast

“…α‐Amylase was assayed at 40°C as described previously [5]. One unit of α‐amylase activity was defined as a decrease in A 595nm of 0.1 after 90 min incubation.…”

“…S. cerevisiae has been widely used to express and study amylases from different eukaryotic organisms and glucoamylases and α‐amylases from different yeast genera have also been analyzed [2–4]. Previously, we isolated two different α‐amylase genes ( SWA1 and SWA2 ) from the yeast Schwanniomyces occidentalis and studied their expression in S. cerevisiae [5–7].…”

Growth Characteristics of Fusants by Protoplast Fusion between the Thermotolerant Yeast, Kluyveromyces marxianus, and the Starch-assimilating Yeast, Schwanniomyces occidentalis

“…Schw. occidentalis produces a-amylase (Abarca et al, 1989;Claros et al, 1993) and glucoamylase (Dowhanick et al, 1990), which play an important role in the consumption of starch residues in Cassava pulp. Schw.…”

“…Schw. occidental's contains the a-amylase genes AMY1 (Abarca et al, 1989) and SWA2 (Claros et al, 1993) and the glucoamylase gene GLA1 (Dowhanick et al, 1990), while there have been no reports that K marxianus possesses an amylase gene. Our preliminary experiments with the polymerase chain reaction using the AMY1, SWA2 and GLA1 primers indicate that the a-amylase genes, AMY1 and SWA2, are absent in the fusants KKU01 and KKUO3 while they are present in Schw.…”