https://ecoliwiki.org/colipedia/api.php?action=feedcontributions&feedformat=atom&user=129.69.146.3EcoliWiki - User contributions [en]2026-11-14T01:05:23ZUser contributionsMediaWiki 1.43.11https://ecoliwiki.org/colipedia/index.php?title=PMID:20148428&diff=1061203PMID:201484282010-03-01T07:23:40Z<p>129.69.146.3: Fill PMID: Page!</p>
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<span style = 'font-size: large'> '''Schneider, S, Gutiérrez, M, Sandalova, T, Schneider, G, Clapés, P, Sprenger, GA and Samland, AK''' (2010) Redesigning the Active Site of Transaldolase TalB from Escherichia coli: New Variants with Improved Affinity towards Nonphosphorylated Substrates.''Chembiochem'' </span><br />
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|{{TableHeadingColor}} |'''Abstract''' || <protect>Recently, we reported on a transaldolase B variant (TalB F178Y) that is able to use dihydroxyacetone (DHA) as donor in aldol reactions. In a second round of protein engineering, we aimed at improving the affinity of this variant towards nonphosphorylated acceptor aldehydes, that is, glyceraldehyde (GA). The anion binding site was identified in the X-ray structure of TalB F178Y where a sulfate ion from the buffer was bound in the active site. Therefore, we performed site-directed saturation mutagenesis at three residues forming the putative phosphate binding site, Arg181, Ser226 and Arg228. The focused libraries were screened for the formation of D-fructose from DHA and d,l-GA by using an adjusted colour assay. The best results with respect to the synthesis of D-fructose were achieved with the TalB F178Y/R181E variant, which exhibited an at least fivefold increase in affinity towards d,l-GA (K(M)=24 mM). We demonstrated that this double mutant can use D-GA, glycolaldehyde and the L-isomer, L-GA, as acceptor substrates. This resulted in preparative synthesis of D-fructose, D-xylulose and L-sorbose when DHA was used as donor. Hence, we engineered a DHA-dependent aldolase that can synthesise the formation of polyhydroxylated compounds from simple and cheap substrates at preparative scale.</protect><br />
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|{{TableHeadingColor}} nowrap |'''Links''' || [http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=20148428 PubMed]<br/>Online version:[http://dx.doi.org/10.1002/cbic.200900720 10.1002/cbic.200900720]<br />
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<references/></div>129.69.146.3https://ecoliwiki.org/colipedia/index.php?title=PMID:20148428&diff=1061202PMID:201484282010-03-01T07:23:38Z<p>129.69.146.3: New PMID: Page!</p>
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