Difference between revisions of "PMID:20585771"
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==Phenotype Annotations== | ==Phenotype Annotations== | ||
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!|Phenotype of!!Taxon Information!!Genotype Information (if known)!!Condition Information!!OMP ID!!OMP Term Name!!ECO ID!!ECO Term Name!!Notes!!Status | !|Phenotype of!!Taxon Information!!Genotype Information (if known)!!Condition Information!!OMP ID!!OMP Term Name!!ECO ID!!ECO Term Name!!Notes!!Status | ||
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| + | a mutation or genetic difference within a strain | ||
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| + | *Taxon: Escherichia coli | ||
| + | *Strain: K-12 | ||
| + | *Substrain: M6 | ||
| + | *NCBI Taxon ID: [http://www.ncbi.nlm.nih.gov/taxonomy?term=83333 83333] | ||
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| + | *Genotype of Reference Strain: gldA | ||
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| + | *Reference Condition: | ||
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| + | Mutation (M6) was able to increase catalytic activity by 26-fold of glycerol dehydrogenase on racemic 1, 3-butadiol for the bioproduction of the pharmaceutical intermediate 4-hydroxy-2-butanone, table 5. | ||
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Revision as of 20:53, 13 December 2011
| Citation |
Zhang, H, Lountos, GT, Ching, CB and Jiang, R (2010) Engineering of glycerol dehydrogenase for improved activity towards 1, 3-butanediol.Appl. Microbiol. Biotechnol. 88:117-24 |
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| Abstract |
The objective of this study was to use protein engineering techniques to enhance the catalytic activity of glycerol dehydrogenase (GlyDH) on racemic 1, 3-butanediol (1, 3-BDO) for the bioproduction of the important pharmaceutical intermediate 4-hydroxy-2-butanone. Three GlyDH genes (gldA) from Escherichia coli K-12, Salmonella enterica, and Klebsiella pneumoniae MGH78578 were shuffled to generate a random mutagenesis library. The nitroblue tetrazolium/phenazine methosulfate high throughput screening protocol was used to select four chimeric enzymes with up to a 2.6-fold improved activity towards 1, 3-BDO. A rational design method was also employed to further improve the enzyme activity after DNA shuffling. Based on the homology model of GlyDH (Escherichia coli), Asp121 was predicted to influence 1, 3-BDO binding and replaced with Ala by site-directed mutagenesis. Combination of the mutations from both DNA shuffling and rational design produced the best mutant with a V (max) value of 126.6 U/mg, a 26-fold activity increase compared with that of the wild type GlyDH from E. coli. |
| Links |
PubMed Online version:10.1007/s00253-010-2735-8 |
| Keywords |
Butylene Glycols; DNA Shuffling; DNA, Bacterial; Escherichia coli K12; Hydroxybutyrates; Klebsiella pneumoniae; Methylphenazonium Methosulfate; Nitroblue Tetrazolium; Protein Engineering; Salmonella enterica; Sugar Alcohol Dehydrogenases |
| edit table |
Main Points of the Paper
Please summarize the main points of the paper.
Materials and Methods Used
Please list the materials and methods used in this paper (strains, plasmids, antibodies, etc).
Phenotype Annotations
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| Phenotype of | Taxon Information | Genotype Information (if known) | Condition Information | OMP ID | OMP Term Name | ECO ID | ECO Term Name | Notes | Status |
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a mutation or genetic difference within a strain |
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Mutation (M6) was able to increase catalytic activity by 26-fold of glycerol dehydrogenase on racemic 1, 3-butadiol for the bioproduction of the pharmaceutical intermediate 4-hydroxy-2-butanone, table 5. |
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Notes
References
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