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Team:WLC-Milwaukee/Modeling
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<h2>yesZ Enzyme Activity</h2> | <h2>yesZ Enzyme Activity</h2> | ||
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| - | + | The kinetic parameters fo the hydrolysis of pNPGal by wild type YesZ Kcat = 81.4 ± 4 s-1, and Km= 3.0 ± 0.2 mM, and Kcat/Km= 27 ± 2 mM -1 s-1. | |
Figure A describes the time dependent inactivation of the YesZ enzyme using DNP2FGal. The time dependent inactivation observed as a single exponential decay to a non-zero value when DNP2FGal was incubated with the enzyme. Studies of inactivation kinetics were performed by pre-incubating 100 microliters of the enzyme at 37 oC with DNP2FGal at a range of concentrations in a total volume of 140 microliters."> | Figure A describes the time dependent inactivation of the YesZ enzyme using DNP2FGal. The time dependent inactivation observed as a single exponential decay to a non-zero value when DNP2FGal was incubated with the enzyme. Studies of inactivation kinetics were performed by pre-incubating 100 microliters of the enzyme at 37 oC with DNP2FGal at a range of concentrations in a total volume of 140 microliters."> | ||
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Revision as of 00:04, 28 September 2013
yesZThe Beta-Galactosidase yesZ is globular and has three aspects that are important to its function. It contains a zinc ion binding site at the C residues 153, 155, and 158 in yellow. The putative nucleophile and acid base cleavage sites are at the E residues 155 and 296 in red. The active site where substrate binding occurs is through the amino acid residue sequence ETSPSYAASL from residues 296- 305. |
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yesZ Enzyme Activity
| The kinetic parameters fo the hydrolysis of pNPGal by wild type YesZ Kcat = 81.4 ± 4 s-1, and Km= 3.0 ± 0.2 mM, and Kcat/Km= 27 ± 2 mM -1 s-1. Figure A describes the time dependent inactivation of the YesZ enzyme using DNP2FGal. The time dependent inactivation observed as a single exponential decay to a non-zero value when DNP2FGal was incubated with the enzyme. Studies of inactivation kinetics were performed by pre-incubating 100 microliters of the enzyme at 37 oC with DNP2FGal at a range of concentrations in a total volume of 140 microliters."> |
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bglSThe endo-1,3-1,4-glucanase bglS is a globular protein that that has two residues of interest. The putative nucleophile and acid-base cleavage sites at the E residues 133 and 137 highlighted in red. |
xynAThe endo-1,4-beta-xylanase xynA is a globular protein that has two residues of interest the nucleophile and acid-base cleavage sites at the E residues 78 and 172 highlighted in red. |
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xynA Enzyme Activity | |
This graph depicts the inhibition of the gene product of XynA found in Bacillus Subtillis Subtillis 168 (BsX) in comparison to the inhibition of the XynA found in Aspergillus Niger (AsX). Sorensen and Sibbensen were observing the inhibitory effects of the TAXI (Triticum Aestivum Xylanase Inhibitor) , specific to Glycoside hydrolase family 11 (GH 11), and XIP (xylanase inhibitor protein), specific to fungal GH 11 but not bacterial GH 11. which XynA is a member. Inhibition was tested with either pure XIP (BsX-XIP and AnX-XIP) or both XIP and TAXI ( BsX-Inhibitor Prep and AnX-Inhibitor Prep. It is evident from this graph that BsX is not effected by XIP but is strongly inhibited by TAXI, causing a decrease in residual xylanase activity by approximately 80%. |
This graph depicts the effect of pH on the interaction between the BsX xylanase and the inhibitor TAXI at a 1:5 concentration. The pH profile of the of the inhibition resembles the ph profile of the enzyme, indicating that TAXI is a competitive inhibitor for the BsX Xylanase. This graph also depicts the optimal pH for the XynA enzyme from Bacillus Subtillis Subtillis 168 to be around 5.5. |
