EC Number   |
Application |
Reference |
|---|
   2.3.1.4 | analysis |
development of a fluorescence assay where reactive thiols of CoA, generated from the acyl transfer reaction, are monitored using monobromobimane |
755991 |
| 2.3.1.4 | biotechnology |
metabolic engineering of Escherichia coli for industrial production of glucosamine and N-acetylglucosamine by overexpression of glucosamine synthase and glucosamine 6-phosphate N-acetyltransferase and inactivation of catabolic genes |
659850 |
| 2.3.1.4 | medicine |
analysis of the global protein expression profiles of lung cancer cell A549 treated with abraxane and paclitaxel. The superior drug effect of abraxane is mainly due to the downregulation of GNA1, which causes proliferative delay and cell adhesion defect |
757009 |
| 2.3.1.4 | molecular biology |
a microtiter plate based assay to detect the GlmU activity is developed: the assay relies on the enzymes MurA and MurB to convert UDP-GlcNAc into UDP-MurNAc with the concomitant reduction of NADPH. When all enzymes and substrates are present NADPH is oxidized with a concomitant decrease in OD340 |
687124 |
| 2.3.1.4 | molecular biology |
an assay for glucosamine-6-phosphate synthase using a yeast glucosamine-6-phosphate N-acetyltransferase 1 (GNA1) as coupling enzyme is developed. The assay measures the production of glucosamine-6-phosphate by either following the consumption of acetyl-CoA spectrophotometrically at 230 nm or quantifying the free thiol with 5,50-dithio-bis(2-nitrobenzoic acid) (Ellmans reagent) in a discontinuous manner. This method is simple to perform and can be adapted to a 96-well microtiter plate format |
684404 |
| 2.3.1.4 | synthesis |
overexpression in Bacillus subtilis for synthesis of N-acetylglucosamine. The expression level of GNA1 is enhanced at the translational level via fusion of an epitope tag to the 5'-terminus of GNA1 gene and ribosome binding site sequence engineering. Enhanced expression of glutamine-fructose-6-phosphate aminotransferaseGlmS is achieved at the transcriptional and translational levels. Under the control of engineered GNA1 and GlmS, the GlcNAc titer and yield in the shake flask increase to 18.5 g/l and 0.37 g GlcNAc/g glucose |
756261 |
| 2.3.1.4 | synthesis |
use of enzyme for synthesis of N-acetylglucosamine in Bacillus subtilis. GNA1 is evolved through error-prone PCR under pyruvate stress to enhance its catalytic activity. Urease from Bacillus paralicheniformis is expressed intracellularly to neutralize the intracellular pH. The activity of mutant CeGNA1 increases by 11.5% at pH 6.5-7.5, with the catalytic efficiency increasing by 27.5% to 1.25 per s and microM. Modulated expression of urease increases the intracellular pH from 6.0 to 6.8. The final engineered produce 25.6 g/l N-acetylglucosamine with a yield of 0.43 g N-acetylglucosamine/g glucose in a shake flask fermentation and produces 82.5 g/l N-acetylglucosamine with a yield of 0.39 g GlcNAc/g glucose by fed-batch fermentation |
757601 |
