GldA, Recombinant, E. coli, aa1-367, His-Tag (Glycerol Dehydrogenase)
Biozol Catalog Number:
USB-351245
Supplier Catalog Number:
351245
Alternative Catalog Number:
USB-351245-20,USB-351245-100
Manufacturer:
US Biological
Category:
Molekularbiologie
GldA catalyzes the NAD-dependent oxidation of glycerol to dihydroxyacetone (glycerone). This protein allows microorganisms to utilize glycerol as a source of carbon under anaerobic conditions. In E. coli, an important role of GldA is also likely to regulate the intracellular level of dihydroxyacetone by catalyzing the reverse reaction, i.e. the conversion of dihydroxyacetone into glycerol. gldA possesses a broad substrate specificity, since it is also able to oxidize 1,2-propanediol and to reduce glycolaldehyde, methylglyoxal and hydroxyacetone into ethylene glycol, lactaldehyde and 1,2-propanediol, respectively. Source: Recombinant protein corresponding to aa1-367 from E. coli gldA fused to His-tag at N-terminal expressed in E. coli. Molecular Weight: ~41.1kD(390aa) Specific Activity: >14Units/ml, one unit will oxidize 1umole of glycerol to dihydroxyacetone/minute at pH 8.0 at 25C Activity Assay: 1. Prepare a 200ul reaction mix into a suitable container: The final concentrations are 93mM glycine, 93mM potassium chloride, 2375mM glycerol, 3mM b-NAD. 2. Equilibrate to 25C and monitor at A340nm until the value is constant using a spectrophotometer. 3. Add 20ul of recombinant gldA protein with various concentrations (0.2ug, 0.1ug, 0.05ug) in 180ul reaction buffer. 4. Mix by inversion and record the decrease at A340nm for 10 minutes. Amino Acid Sequence: MGSSHHHHHH SSGLVPRGSH MGSMDRIIQS CSQNEIDRLR GIAETAQCGA ILGIGGGKTL AGAPARLLAA GIGDALATWF EARACSRSGA GGLAAAHAVH NGLTAIPDAH HYYHGEKVAF NMPGGATPDQ VYAALLVADQ YGQRFLQEWE PGKYIQGADV INRLGEYLKP LAERWLVVGD KFVLGFAQST VEKSFKDAGL VVEIAPFGGE DTAKALAHFM GVPVAIAPTI ASTDAPCSAL SVIYTDEGEF DRYLLLPNNP NMVIVDTKIV TTMAGGKCTQ AALALAELCY NTLLEEGEKA MLAAEQHVVT PALERVIEAN TYLSGVGFES GTLTQLVLEN APVEEIETVA ALSHAVGLPI TLAQLDIKED VPAKMRIVAE AACAEGETIH Storage and Stability: May be stored at 4C for short-term only. Aliquot to avoid repeated freezing and thawing.. Store at -20C. Aliquots are stable for 6 months after receipt at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
