EC Number |
Protein Variants |
Reference |
---|
2.6.1.16 | K595H |
site-directed mutagenesis, the mutant shows unaltered activity (5 U/l) compared to wild-type (5 U/l) |
-, 758961 |
2.6.1.16 | L468P |
mutant shows increased resistance against glucosamine-6-phosphate |
672444 |
2.6.1.16 | L593S |
site-directed mutagenesis, the mutant shows highly increased activity (48 U/l) compared to wild-type (5 U/l) |
-, 758961 |
2.6.1.16 | more |
combinatorial fine-tuning of glucosamine-6-phosphate N-acetyltransferase (GNA1) and glutamine-fructose-6-phosphate aminotransferase (GlmS) expression by 5-terminus fusion engineering leads to overproduction of N-acetylglucosamine in Bacillus subtilis. Enhanced expression of GlmS is achieved at the transcriptional and translational levels by fusing an mRNA stabilizer to the 5'-terminus of GlmS gene. Under the control of GNA1 (fusion with cMyc tag and with the optimum RBS M-Rm) and GlmS (fusion with mRNA stabilizer DELTAermC+14/7A), the GlcNAc titer and yield in the shake flask increase to 18.5 g/l and 0.37 g GlcNAc/g glucose, which are 2.9fold and 2.3fold that of the control, respectively. Synthetic pathway fine-tuning method at the transcriptional and translational levels by combinatorial modulation of regulatory elements, including epitope tag, RBS sequence, and mRNA stabilizer, method evaluation, overview |
-, 756261 |
2.6.1.16 | more |
Crispr-mediated elimination of Drosophila gfat genes and generation of single gene null mutations in the fly counterparts of gfat1 and gfat2 (gfat1-/- and gfat2-/-). Deletions for either enzyme are fully lethal and homozygotes lacking either GFAT1 or GFAT2 die at or prior to the first instar larval stage. When genetically eliminated, neither isoform is able to compensate for the other. Dietary supplementation with D-glucosamine-6-phosphate rescues GFAT2 deficiency and restores viability to gfat2-/- mutants. In contrast, glucosamine-6-phosphate does not rescue gfat1-/- animals |
759211 |
2.6.1.16 | more |
expression of truncated enzyme variants as His-tagged proteins. Fragments encompassing residues 1-345 and 346-712 represent the functional glutamine amide-hydrolysing GAH and hexose phosphate-isomerizing domains ISOM, respectively. The native GAH domain is monomeric, whereas the native ISOM domain forms tetramers, as does the whole enzyme. The binding site for the feedback inhibitor, uridine 5'-diphospho-N-acetyl-D-glucosamine, is located in the ISOM domain. Inhibitor binding affects amidohydrolysing activity of the GAH domain and, as a consequence, the D-glucosamine-6-phosphate-synthetic activity of the whole enzyme. The fragment containing residues 218-283 is neither involved in ligand binding nor in protein oligomerization. An intramolecular channel connects the GAH and ISOM domains. The channel becomes leaky upon deletion of amino acids 709-712 and in the W97F and W97G mutants |
671896 |
2.6.1.16 | more |
improvement of enzyme BsGlms activity through computer simulation and site saturation mutagenesis, method, overview |
-, 758961 |
2.6.1.16 | more |
three recombinant versions containing internal oligoHis fragments are constructed: (a) by substituting residues 343-348 of the interdomain undecapeptide linker with hexaHis, (b) by replacing solvent-exposed residues 655-660 of the isomerase domain with hexaHis, and (c) by replacing amino acids at positions 568 and 569 with His residues to generate the three-dimensional hexaHis microdomain in the enzyme quaternary structure. The resulting constructs are purified to homogeneity. Catalytic properties are comparable with that of the wild-type enzyme. The construct containing the 655-660 hexaHis insert is found to be a homodimeric protein |
723015 |
2.6.1.16 | S243E |
increase in activity, 2fold lower Km value for D-fructose 6-phosphate than wild-type |
685109 |
2.6.1.16 | S449P |
mutant shows increased resistance against glucosamine-6-phosphate |
672444 |