BMB464_FinalPaper - Extraction and purification...

Info iconThis preview shows pages 1–3. Sign up to view the full content.

View Full Document Right Arrow Icon
Extraction and purification of α-Galactosidase from Saccharomyces pastorianus From the Department of Biochemistry, Microbiology and Molecular Biology (BMMB) at the University of Maine in Orono Alpha-galactosidase catalyzes the hydrolysis of sugars containing the 1-4 galactosidic bonds and 1-6 terminal galactosyl groups. Since alpha-galactosidase is present in a wide range of plants, animals, and microorganisms. Alpha-galactosidase was collected from Saccharomyces pastorianus , a top-fermenting brewer’s yeast. It was purified by a combination of ion-exchange resin, NaCl gradient elution and dialysis. The starting specific activity was 0.014 and the final specific activity was 0.2 which showed a purification factor of 14.29. The purified alpha-galactosidase was analyzed by SDS polyacrylamide gel electrophoresis and native gel electrophoresis. The molecular weight was estimated to be 110kDa. Kinetic analysis of the alpha-galactosidase showed that the optimal pH and temperature that alpha-galactosidase worked at was pH 4.5 and a temperature of ~40 º C). The enzyme was also subjected to different inhibitors (glucose and galactose) to see how these compounds interfered with the activity of alpha- galactosidase. All these data showed that the isolation and characterization of alpha- galactosidase is achievable from the S. pastorianus yeast species. Alpha-galactosidase will continue to be an important enzyme commercially and the understanding of it can only help to open new markets and innovations. Alpha-galactosidase is an enzyme that hydrolyzes α 1, 6-galactosidic bonds. The a-1,6-galactosidic bonds can be found in a number of carbohydrates such as stachyose raffinose, and verbascose; carbohydrates found in bacteria, plants, and animals. Raffinose
Background image of page 1

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full Document Right Arrow Icon
is hydrolyzed by α-galactosidase into D-galactose, stachyose and melibiose both hydrolzies to D-galactose. All three of these sugars also yeild sucrose when hydrolyzed. Alpha -galactosidase has several commercial applications. The first and best known is the product Beano. The α-galactosidase in this product is derived from the fungus Aspergillus niger (6). Humans cannot digest stachyose due to its a-1,6- galactosidic bonds and the lack of an enzyme capable of cleaving the bonds. Hence, stachyose passes through the small intestine without being absorbed. When it enters the large intestine, the normal anaerobic microbial flora ferments the stachyose creating methane and carbon dioxide (6). With the addition of the α-galactosidase prior to eating, stachyose can be broken down into monosaccharides that can be absorbed by the small intestine, thereby depriving the normal flora the carbohydrate it needs to ferment. A second commercial use for α-galactosidase is in the production of beet sugars. By hydrolyzing the a-1,6-galactosidic bonds in raffinose (found in beet sugar), the sugar is converted into galactose and sucrose, which help to expedite crystallization. Another use of
Background image of page 2
Image of page 3
This is the end of the preview. Sign up to access the rest of the document.

{[ snackBarMessage ]}

Page1 / 23

BMB464_FinalPaper - Extraction and purification...

This preview shows document pages 1 - 3. Sign up to view the full document.

View Full Document Right Arrow Icon
Ask a homework question - tutors are online