Group:SMART:2010 Pingry SMART Team: Difference between revisions

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'''Design description'''

The conformation of 2.5-DKGR A is a parallel alpha/beta barrel of eight <scene name='2010_Pingry_SMART_Team/1a80-default/2'>alpha helices(highlighted red) and eight beta strands(highlighted blue).</scene> ~~Notably, the~~ alpha-beta structural motif is ~~common~~ among other enzymes of the aldo-keto reductase family. In addition, the residue,<scene name='2010_Pingry_SMART_Team/1a80-original/10'>Tyr50</scene>, is conserved in all aldo-keto-reductases and functions as both part of the catalytic triad ~~and proton donor of aldo-keto reductases~~. The residues,<scene name='2010_Pingry_SMART_Team/1a80-original/7'>Ala47 and Trp77;</scene>, are conserved in all AKR (aldo-keto reductase) enzymes as well as contributors to the formation of the active-site pocket. Located at the C-terminal side of the barrel is the active site for the

The conformation of 2.5-DKGR A is a parallel alpha/beta barrel of eight <scene name='2010_Pingry_SMART_Team/1a80-default/2'>alpha helices(highlighted red) and eight beta strands(highlighted blue).</scene> This alpha-beta structural motif is demonstrated among other enzymes of the aldo-keto reductase family. In addition, the residue,<scene name='2010_Pingry_SMART_Team/1a80-original/10'>Tyr50</scene>, is also conserved in all aldo-keto-reductases and functions as both the proton donor and part of the catalytic triad. The residues,<scene name='2010_Pingry_SMART_Team/1a80-original/7'>Ala47 and Trp77;</scene>, are conserved in all AKR (aldo-keto reductase) enzymes as well as contributors to the formation of the active-site pocket. Located at the C-terminal side of the barrel is the active site for the

<scene name='2010_Pingry_SMART_Team/1a80-original/12'>NADPH cofactor(shown in wireframe and colored CPK)</scene> to bind to 2.5-DKGR A. The NADPH cofactor is stabilized through hydrogen bonds, ionic bonds, and an aromatic pi-stacking interaction between <scene name='2010_Pingry_SMART_Team/1a80-original/2'>Trp187</scene> and the nicotinamide ring of NADPH . Although 2,5-DKGR A functions with NADPH as a cofactor, NADH is preferred for a more efficient production of vitamin C. To achieve this, mutations of the original side chains of <scene name='2010_Pingry_SMART_Team/1a80-default/1'>Lys232, Phe22, Arg238, Ala272</scene> were conducted. Significantly, the<scene name='2010_Pingry_SMART_Team/1a80-original/8'>Lys232, Ser233, and Val234</scene> sidechain interact with the phosphate group of NADPH. In order to acommodate for the cofactor, NADH, and the absent phosphate group, these side chains have been modified in the mutant form.

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 '''Design description'''
-The conformation of 2.5-DKGR A is a parallel alpha/beta barrel of eight <scene name='2010_Pingry_SMART_Team/1a80-default/2'>alpha helices(highlighted red) and eight beta strands(highlighted blue).</scene> Notably, the alpha-beta structural motif is common among other enzymes of the aldo-keto reductase family. In addition, the residue,<scene name='2010_Pingry_SMART_Team/1a80-original/10'>Tyr50</scene>, is conserved in all aldo-keto-reductases and functions as both part of the catalytic triad and proton donor of aldo-keto reductases. The residues,<scene name='2010_Pingry_SMART_Team/1a80-original/7'>Ala47 and Trp77;</scene>, are conserved in all AKR (aldo-keto reductase) enzymes as well as contributors to the formation of the active-site pocket. Located at the C-terminal side of the barrel is the active site for the
+The conformation of 2.5-DKGR A is a parallel alpha/beta barrel of eight <scene name='2010_Pingry_SMART_Team/1a80-default/2'>alpha helices(highlighted red) and eight beta strands(highlighted blue).</scene> This alpha-beta structural motif is demonstrated among other enzymes of the aldo-keto reductase family. In addition, the residue,<scene name='2010_Pingry_SMART_Team/1a80-original/10'>Tyr50</scene>, is also conserved in all aldo-keto-reductases and functions as both the proton donor and part of the catalytic triad. The residues,<scene name='2010_Pingry_SMART_Team/1a80-original/7'>Ala47 and Trp77;</scene>, are conserved in all AKR (aldo-keto reductase) enzymes as well as contributors to the formation of the active-site pocket. Located at the C-terminal side of the barrel is the active site for the
 <scene name='2010_Pingry_SMART_Team/1a80-original/12'>NADPH cofactor(shown in wireframe and colored CPK)</scene> to bind to 2.5-DKGR A. The NADPH cofactor is stabilized through hydrogen bonds, ionic bonds, and an aromatic pi-stacking interaction between <scene name='2010_Pingry_SMART_Team/1a80-original/2'>Trp187</scene> and the nicotinamide ring of NADPH . Although 2,5-DKGR A functions with NADPH as a cofactor, NADH is preferred for a more efficient production of vitamin C. To achieve this, mutations of the original side chains of <scene name='2010_Pingry_SMART_Team/1a80-default/1'>Lys232, Phe22, Arg238, Ala272</scene> were conducted. Significantly, the<scene name='2010_Pingry_SMART_Team/1a80-original/8'>Lys232, Ser233, and Val234</scene> sidechain interact with the phosphate group of NADPH. In order to acommodate for the cofactor, NADH, and the absent phosphate group, these side chains have been modified in the mutant form.