4ZXZ
Crystal structure of a highly thermal stable but inactive levoglucosan kinase.
Summary for 4ZXZ
| Entry DOI | 10.2210/pdb4zxz/pdb |
| Descriptor | Levoglucosan kinase (2 entities in total) |
| Functional Keywords | transferase, sugar kinase., de novo protein |
| Biological source | Lipomyces starkeyi (Oleaginous yeast) |
| Total number of polymer chains | 2 |
| Total formula weight | 98917.88 |
| Authors | Bacik, J.P.,Klesmith, J.R.,Whitehead, T.A. (deposition date: 2015-05-20, release date: 2015-09-23, Last modification date: 2023-09-27) |
| Primary citation | Klesmith, J.R.,Bacik, J.P.,Michalczyk, R.,Whitehead, T.A. Comprehensive Sequence-Flux Mapping of a Levoglucosan Utilization Pathway in E. coli. Acs Synth Biol, 4:1235-1243, 2015 Cited by PubMed Abstract: Synthetic metabolic pathways often suffer from low specific productivity, and new methods that quickly assess pathway functionality for many thousands of variants are urgently needed. Here we present an approach that enables the rapid and parallel determination of sequence effects on flux for complete gene-encoding sequences. We show that this method can be used to determine the effects of over 8000 single point mutants of a pyrolysis oil catabolic pathway implanted in Escherichia coli. Experimental sequence-function data sets predicted whether fitness-enhancing mutations to the enzyme levoglucosan kinase resulted from enhanced catalytic efficiency or enzyme stability. A structure of one design incorporating 38 mutations elucidated the structural basis of high fitness mutations. One design incorporating 15 beneficial mutations supported a 15-fold improvement in growth rate and greater than 24-fold improvement in enzyme activity relative to the starting pathway. This technique can be extended to improve a wide variety of designed pathways. PubMed: 26369947DOI: 10.1021/acssynbio.5b00131 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.2 Å) |
Structure validation
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