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

Xanthophyllomyces dendrorhous grown in different media shows amylolytic activity, consisting in an extracellular exo-acting enzyme able to hydrolysed α,1-4 glycosidic bonds from soluble starch, which also cleaves maltose and malto-oligosaccharides. The enzyme was purified, using basically a couple of chromatography process on DEAE-Sephacel. It is a glycoprotein with a molecular weight estimated to be 60.2 kDa based on its mobility in SDS-PAGE and 115 kDa based on gel filtration. N-linked carbohydrate accounts for 12% of the total mass. It exhibited optimum activity at pH 5.5 and 45 • C. Thermostability analysis indicated that it was stable to thermal treatment up to 50 • C; 50% of the activity was maintained after 3 h. The rate parameters measured for the hydrolysis of starch and various chain length malto-oligosaccharides shows high catalytic efficiency, calculated by the relationship V cat /K m , for malto-oligosaccharides, such as maltotriose (873 mM −1 min −1 ), or maltoheptose (698 mM −1 min −1 ). The new enzyme hydrolysed soluble starch with nearly 3.5-and 1.4-fold lower efficiency than that for maltotriose and maltose, respectively. No activity was found on heterogeneous substrates, such as sucrose and aryl α-glucoside, or on isomalto-oligosaccharides. In accordance to substrate specificity profile, the new enzyme was classified as an α-glucosidase.

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Purification and biochemical characterization of an α‐glucosidase from Xanthophyllomyces dendrorhous

Marín

Linde

Lobato

2006

Yeast

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Current Awareness

2001

Yeast

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In order to keep subscribers up‐to‐date with the latest developments in their field, this current awareness service is provided by John Wiley & Sons and contains newly‐published material on yeasts. Each bibliography is divided into 10 sections. 1 Books, Reviews & Symposia; 2 General; 3 Biochemistry; 4 Biotechnology; 5 Cell Biology; 6 Gene Expression; 7 Genetics; 8 Physiology; 9 Medical Mycology; 10 Recombinant DNA Technology. Within each section, articles are listed in alphabetical order with respect to author. If, in the preceding period, no publications are located relevant to any one of these headings, that section will be omitted. (6 weeks journals ‐ search completed 17th. Oct. 2001)

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Kang

Park

Chin³

et al. 2003

Biotechnology Letters

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The gene encoding Schwanniomyces occidentalis alpha-amylase (AMY) was introduced into the chromosomal delta sequences of an industrial strain of Saccharomyces cerevisiae. To obtain a strain suitable for commercial use, an delta-integrative cassette devoid of bacterial DNA sequences was constructed that contains the AMY gene and aureobasidin A resistance gene (AUR1-C) as the selection marker. The AMY gene was expressed under the control of the alcohol dehydrogenase gene promoter (ADC1p). The alpha-amylase activity of Sacc. cerevisiae transformed with this integrative cassette was 6 times higher than that of Sch. occidentalis. The transformants (integrants) were mitotically stable after 100 generations in nonselective medium.

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

Cited by 7 publications

References 21 publications

Purification and biochemical characterization of an α‐glucosidase from Xanthophyllomyces dendrorhous

Purification and biochemical characterization of an α‐glucosidase from Xanthophyllomyces dendrorhous

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