Candida albicans SOU1 encodes a sorbose reductase required for L-sorbose Utilization

Previous work in our laboratory showed that L-sorbose utilization in Candida albicans is subject to a novel form of regulation which involves a reversible increase or decrease in the copy number of chromosome 5. Furthermore, the structural gene SOU1 is required for L-sorbose utilization and encodes...

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Main Authors: Greenberg, J., Price, N., Oliver, Richard, Sherman, F., Rustchenko, E.
Format: Journal Article
Published: 2005
Online Access:http://hdl.handle.net/20.500.11937/17830
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author Greenberg, J.
Price, N.
Oliver, Richard
Sherman, F.
Rustchenko, E.
author_facet Greenberg, J.
Price, N.
Oliver, Richard
Sherman, F.
Rustchenko, E.
author_sort Greenberg, J.
building Curtin Institutional Repository
collection Online Access
description Previous work in our laboratory showed that L-sorbose utilization in Candida albicans is subject to a novel form of regulation which involves a reversible increase or decrease in the copy number of chromosome 5. Furthermore, the structural gene SOU1 is required for L-sorbose utilization and encodes a member of the short chain dehydrogenase family. However, the precise function of SOU1 was not known and neither was the pathway for L-sorbose utilization. We have now expressed SOU1 at a high level from a replicative plasmid having a constitutive ADH1 promoter and purified a version of Sou1p tagged with the FLAG epitope at the N-terminus. Sou1FLAGNp has a sorbose reductase activity which utilizes NADPH as a co-factor and converts L-sorbose to D-sorbitol. It can also less efficiently utilize fructose as a substrate with NADPH as a co-factor, converting fructose to mannitol. In agreement with prediction, the purified enzyme has a subunit molecular weight of 31 kDa and a pI of about 4.8. It probably consists of four identical subunits and has a pH optimum of 6.2. The L-sorbose utilization pathway in C. albicans probably converts L-sorbose to fructose-6-phosphate via D-sorbitol as an intermediate. The first step is catalysed by Sou1p. We also found that C. albicans extracts have a D-sorbitol-6-phosphate dehydrogenase activity, not encoded by SOU1, which utilizes NADP as a co-factor. This activity has not been described previously in yeasts and may be involved in the conversion of phosphorylated D-sorbitol to fructose-6-phosphate or glucose-6-phosphate.
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spelling curtin-20.500.11937-178302017-09-13T15:42:43Z Candida albicans SOU1 encodes a sorbose reductase required for L-sorbose Utilization Greenberg, J. Price, N. Oliver, Richard Sherman, F. Rustchenko, E. Previous work in our laboratory showed that L-sorbose utilization in Candida albicans is subject to a novel form of regulation which involves a reversible increase or decrease in the copy number of chromosome 5. Furthermore, the structural gene SOU1 is required for L-sorbose utilization and encodes a member of the short chain dehydrogenase family. However, the precise function of SOU1 was not known and neither was the pathway for L-sorbose utilization. We have now expressed SOU1 at a high level from a replicative plasmid having a constitutive ADH1 promoter and purified a version of Sou1p tagged with the FLAG epitope at the N-terminus. Sou1FLAGNp has a sorbose reductase activity which utilizes NADPH as a co-factor and converts L-sorbose to D-sorbitol. It can also less efficiently utilize fructose as a substrate with NADPH as a co-factor, converting fructose to mannitol. In agreement with prediction, the purified enzyme has a subunit molecular weight of 31 kDa and a pI of about 4.8. It probably consists of four identical subunits and has a pH optimum of 6.2. The L-sorbose utilization pathway in C. albicans probably converts L-sorbose to fructose-6-phosphate via D-sorbitol as an intermediate. The first step is catalysed by Sou1p. We also found that C. albicans extracts have a D-sorbitol-6-phosphate dehydrogenase activity, not encoded by SOU1, which utilizes NADP as a co-factor. This activity has not been described previously in yeasts and may be involved in the conversion of phosphorylated D-sorbitol to fructose-6-phosphate or glucose-6-phosphate. 2005 Journal Article http://hdl.handle.net/20.500.11937/17830 10.1002/yea.1282 restricted
spellingShingle Greenberg, J.
Price, N.
Oliver, Richard
Sherman, F.
Rustchenko, E.
Candida albicans SOU1 encodes a sorbose reductase required for L-sorbose Utilization
title Candida albicans SOU1 encodes a sorbose reductase required for L-sorbose Utilization
title_full Candida albicans SOU1 encodes a sorbose reductase required for L-sorbose Utilization
title_fullStr Candida albicans SOU1 encodes a sorbose reductase required for L-sorbose Utilization
title_full_unstemmed Candida albicans SOU1 encodes a sorbose reductase required for L-sorbose Utilization
title_short Candida albicans SOU1 encodes a sorbose reductase required for L-sorbose Utilization
title_sort candida albicans sou1 encodes a sorbose reductase required for l-sorbose utilization
url http://hdl.handle.net/20.500.11937/17830