5.1.3.8: N-acylglucosamine 2-epimerase
This is an abbreviated version!
For detailed information about N-acylglucosamine 2-epimerase, go to the full flat file.
Word Map on EC 5.1.3.8
-
5.1.3.8
-
n-acetylneuraminic
-
neu5ac
-
n-acetyl-d-neuraminic
-
anabaena
-
mannac
-
takahashi
-
n-acetyl-d-mannosamine
-
renbp
-
nal
-
n-acetylmannosamine
-
synthesis
-
biotechnology
- 5.1.3.8
-
n-acetylneuraminic
- neu5ac
-
n-acetyl-d-neuraminic
- anabaena
- mannac
-
takahashi
- n-acetyl-d-mannosamine
-
renbp
- nal
- n-acetylmannosamine
- synthesis
- biotechnology
Reaction
Synonyms
Acylglucosamine 2-epimerase, AGE, AGE2, anAGE, AvaAGE, bage, bGlcNAc 2-epimerase, Epimerase, acylglucosamine 2-, GlcNAc 2-epimerase, GlcNAc-2-epimerase, N-acetyl-D-glucosamine 2-epimerase, N-acetyl-D-glucosamine-2-epimerase, N-Acetylglucosamine 2-epimerase, N-acyl-D-glucosamine 2-epimerase, N-acylglucosamine 2-epimerase, PhGn2E, renin binding protein, Renin-binding protein, RNBP
ECTree
5.1.3.8 N-acylglucosamine 2-epimeraseAdvanced search results
results (
results (Engineering
Engineering on EC 5.1.3.8 - N-acylglucosamine 2-epimerase
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
top
PROTEIN VARIANTS 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
C125S
C125S/C210S
-
mutant enzyme shows 54.8% of the activity relative to the wild-type enzyme
C125S/C210S/C239S
-
mutant enzyme shows 49.3% of the activity relative to the wild-type enzyme
C125S/C210S/C239S/C203S
-
mutant enzyme shows 28.7% of the activity relative to the wild-type enzyme
C125S/C210S/C239S/C203S/C386S
-
mutant enzyme shows 23.8% of the activity relative to the wild-type enzyme
C125S/C386S
-
mutant enzyme shows 68.7% of the activity relative to the wild-type enzyme
C125S/C390S
-
mutant enzyme shows 5.1% of the activity relative to the wild-type enzyme
C210S
-
relative specific activity in the extract is nearly the same to that of the wild-type enzyme
C210S/C386S
-
mutant enzyme shows 88.7% of the activity relative to the wild-type enzyme
C210S/C390S
-
mutant enzyme shows 30.9% of the activity relative to the wild-type enzyme
C239S
-
relative specific activity in the extract is nearly the same to that of the wild-type enzyme
C239S/C386S
-
mutant enzyme shows 116% of the activity relative to the wild-type enzyme
C239S/C390S
-
mutant enzyme shows 27.8% of the activity relative to the wild-type enzyme
C302S
-
relative specific activity in the extract is nearly the same to that of the wild-type enzyme
C302S/C386S
-
mutant enzyme shows 65.8% of the activity relative to the wild-type enzyme
C302S/C390S
-
mutant enzyme shows 7.4% of the activity relative to the wild-type enzyme
C386S
C390S
C41S
C41S/C125S
-
mutant enzyme shows 17.7% of the activity relative to the wild-type enzyme
C41S/C125S/C210S
-
mutant enzyme shows 28.1% of the activity relative to the wild-type enzyme
C41S/C125S/C210S/C239S
-
mutant enzyme shows 9.7% of the activity relative to the wild-type enzyme
C41S/C386S
-
mutant enzyme shows 0.7% of the activity relative to the wild-type enzyme
C66S
C66S/C125S
-
mutant enzyme shows 39.4% of the activity relative to the wild-type enzyme
C66S/C125S/C210S
-
mutant enzyme shows 23.9% of the activity relative to the wild-type enzyme
C66S/C125S/C210S/C239S
-
mutant enzyme shows 58.4% of the activity relative to the wild-type enzyme
C66S/C125S/C210S/C239S/C302S
-
mutant enzyme shows 15.5% of the activity relative to the wild-type enzyme
C66S/C386S
-
mutant enzyme shows 113% of the activity relative to the wild-type enzyme
C66S/C390S
-
mutant enzyme shows 14.4% of the activity relative to the wild-type enzyme
DELTA400-417
-
C-terminal deletion mutant has approximately 50% activity relative to the wild-type enzyme
K151V
site-directed mutagenesis, the mutation within the ATP-binding site leads to biphasic enzyme inactivation in the presence of 400 mM pyruvate
K155A
site-directed mutagenesis, the mutation within the ATP-binding site leads to biphasic enzyme inactivation in the presence of 400 mM pyruvate
K157L
site-directed mutagenesis, the mutation within the ATP-binding site leads to biphasic enzyme inactivation in the presence of 400 mM pyruvate
K160I
site-directed mutagenesis, within the ATP-binding site, the mutant shows no inactivation by pyruvate, in contrast to the wild-type enzyme, but significantly impaired kinetic parameters compared to wild-type
K160L
site-directed mutagenesis, within the ATP-binding site, the mutant shows no inactivation by pyruvate, in contrast to the wild-type enzyme, but significantly impaired kinetic parameters compared to wild-type
K160N
site-directed mutagenesis, within the ATP-binding site, the mutant shows no inactivation by pyruvate, in contrast to the wild-type enzyme, but significantly impaired kinetic parameters compared to wild-type
additional information
C125S
-
relative specific activity in the extract is nearly the same to that of the wild-type enzyme
C386S
-
relative specific activity in the extract is nearly the same to that of the wild-type enzyme
C390S
-
relative specific activity in the extract is nearly the same to that of the wild-type enzyme
C41S
-
relative specific activity in the extract is nearly the same to that of the wild-type enzyme
C66S
-
relative specific activity in the extract is nearly the same to that of the wild-type enzyme
additional information
metabolic channeling enables efficient transfer of the intermediates by forming a multienzyme complex. To leverage the metabolic channeling for improved biosynthesis, N-acetylneuraminic acid lyase from Corynebacterium glutamicum ATCC 13032 (CgNal, EC 4.1.3.3) and N-acetylglucosamine-2-epimerase from Anabaena sp. CH1 (anAGE) are coexpressed in Escherichia coli and the whole cell are used to synthesize N-acetylneuraminic acid (Neu5Ac) from N-acetylglucosamine (GlcNAc) and pyruvate. To get the multienzyme complex, a polycistronic plasmid with high levels of CgNal and anAGE expression is constructed by tuning the orders of the genes. The Shine-Dalgarno (SD) sequence and aligned spacing (AS) distance are optimized. The Escherichia coli strain Rosetta harboring the polycistronic plasmid pET-28a-SD2-AS1-CgNal-SD-AS-anAGE increases the production of Neu5Ac by 58.7% to 92.5 g/l in 36 h by whole-cell catalysis and by 21.9% up to 112.8 g/l in 24 h with the addition of Triton X-100
additional information
production of N-acetyl-D-neuraminic acid by recombinant single whole cells co-expressing N-acetyl-D-glucosamine-2-epimerase and N-acetyl-D-neuraminic acid aldolase in Escherichia coli. Method development and evaluation, overview
additional information
-
construction of a series of chimeric enzymes successively replacing the three domains of the human enzyme - N-terminal, middle, and C-terminal - with the corresponding domains of the rat enzyme. Chimerae are expressed in Escherichia coli JM109 under the control of the Taq promoter. Chimeric enzymes of HHR, RHH and RHR - where H is a human type domain and R is a rat type domain - have nearly the same nucleotide specificity as the human enzyme. HRR, HRH, and RRH chimeras have the same nucleotide specificity as the rat enzyme. These results indicate that the middle domain of the enzyme molecule participates in the specificity for and binding of nucleotides
additional information
-
mutational analysis of multi-cysteine/serine mutants reveals that Cys41 and Cys390 are critical for the activity or stabilization of the enzyme, while cysteine residues in the middle of the enzyme, Cys125, Cys210, Cys239, and Cys302 have no essential function in relation to the activity
additional information
-
construction of a series of chimeric enzymes successively replacing the three domains of the human enzyme - N-terminal, middle, and C-terminal - with the corresponding domains of the rat enzyme. Chimeras are expressed in Escherichia coli JM109 under the control of the Taq promoter. Chimeric enzymes of HHR, RHH and RHR - where H is a human type domain and R is a rat type domain - have nearly the same nucleotide specificity as the human enzyme. HRR, HRH, and RRH chimeras have the same nucleotide specificity as the rat enzyme. These results indicate that the middle domain of the enzyme molecule participates in the specificity for and binding of nucleotides
additional information
the majority of biocatalytic approaches for Neu5Ac synthesis involve an N-acylglucosamine 2-epimerase (AGE, EC 5.3.1.8) in combination with an N-acetylneuraminate lyase (NAL, EC 4.1.3.3)
additional information
two-step enzymatic synthesis of N-acetylneuraminic acid (Neu5Ac) using an N-acyl-D-glucosamine 2-epimerase from Anabaena variabilis ATCC 29413 (AvaAGE) in combination with an N-acetylneuraminate lyase (NAL) from Escherichia coli. AvaAGE epimerizes N-acetyl-D-glucosamine (GlcNAc) to N-acetyl-D-mannosamine (ManNAc), which then reacts with pyruvate in a NAL-catalyzed aldol condensation to form Neu5Ac. AvaAGE is inactivated by high pyruvate concentrations, which are used to push the NAL reaction toward the product side. A biphasic inactivation is observed in the presence of 50-800 mM pyruvate resulting in activity losses of the AvaAGE of up to 60% within the first hour. Site-directed mutagenesis reveals that pyruvate modifies one of the four lysine residues in the ATP-binding site of AvaAGE
