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Literature summary for 2.4.1.25 extracted from
- Identification of crucial amino acid residues involved in large ring cyclodextrin synthesis by amylomaltase from Corynebacterium glutamicum (2023), Comput. Struct. Biotechnol. J., 21, 899-909.
Application
| Application | Comment | Organism |
|---|---|---|
| synthesis | amylomaltase can be used to synthesize large ring cyclodextrins (LR-CDs), applied as drug solubilizer, gene delivery vehicle and protein aggregation suppressor. The enzyme has the ability to synthesize highly sought cyclic glucans, or large ring cyclodextrins (LR-CDs), with a degree of polymerization (DP) of more than eight through intramolecular transglycosylation reaction. The LR-CDs are highly soluble in water and relatively have a larger hydrophobic cavity compared to the small ring cyclodextrin | Corynebacterium glutamicum |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3). Non-fusion and His-tagged wild-type CgAM show similar specific activity on starch transglycosylation | Corynebacterium glutamicum |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| A406V | site-directed mutagenesis, the mutant shows higher thermostability and gives higher amount of LR-CD products, in comparison to the wild-type enzyme | Corynebacterium glutamicum |
| A413F | site-directed mutagenesis, the mutant produces larger LR-CDs from CD36-CD40 as compared to CD29 by the wild-type, but with low yield. The A413F mutation affects the enzyme activities: starch tranglycosylation, disproportionation and cyclization | Corynebacterium glutamicum |
| E231Y | site-directed mutagenesis, the E231Y mutant exhibits much higher kcat and Km values for starch transglycosylation, compared to that of the wild-type | Corynebacterium glutamicum |
| G417F | site-directed mutagenesis, the mutant produces larger LR-CDs from CD36-CD40 as compared to CD29 by the wild-type, but with low yield. The G417F mutation hinders the enzyme's cyclization activity | Corynebacterium glutamicum |
| additional information | generation of truncated mutant DELTA167 that exhibits no starch transglycosylation activity, indicating that the N-terminal domain of CgAM is necessary for enzyme activity. Mutants molecular dynamic simulations, overview. The wild-type enzyme exhibits cyclization specific activity at 0.0012 U/mg, while P228Y, E231Y and G417F mutants show the same level of specific activity at range 0.0007 0.0013 U/mg | Corynebacterium glutamicum |
| N287Y | site-directed mutagenesis, the mutant exhibits a higher thermostability, a changed LR-CD profile, and an increased substrate preference for maltoheptaose (G5) compared to wild-type | Corynebacterium glutamicum |
| P228Y | site-directed mutagenesis, the mutant produces larger LR-CDs from CD36-CD40 as compared to CD29 by the wild-type. The P228Y mutation significantly lowers the kcat of disproportionation activity compared to wild-type | Corynebacterium glutamicum |
| Y172A | site-directed mutagenesis, the mutation causes a shift of principal LR-CDs to the larger size products | Corynebacterium glutamicum |
| Y23A | site-directed mutagenesis, the mutation affects the kinetic parameters of starch transglycosylation and cyclization | Corynebacterium glutamicum |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | Michaelis-Menten kinetic analysis | Corynebacterium glutamicum | |
| 0.0718 | - |
starch | pH 6.0, 30°C, starch transglycosylation, recombinant mutant G417F | Corynebacterium glutamicum |  |
| 0.0741 | - |
starch | pH 6.0, 30°C, starch transglycosylation, recombinant mutant P228Y | Corynebacterium glutamicum | |
| 0.0822 | - |
starch | pH 6.0, 30°C, starch transglycosylation, recombinant wild-type enzyme | Corynebacterium glutamicum | |
| 0.14 | - |
starch | pH 6.0, 30°C, starch transglycosylation, recombinant mutant Y23A | Corynebacterium glutamicum | |
| 0.186 | - |
starch | pH 6.0, 30°C, starch transglycosylation, recombinant mutant E231Y | Corynebacterium glutamicum | |
| 12.27 | - |
starch | pH 6.0, 30°C, disproportionation activity, recombinant mutant Y23A | Corynebacterium glutamicum | |
| 15.4 | - |
starch | pH 6.0, 30°C, disproportionation activity, recombinant mutant E231Y | Corynebacterium glutamicum | |
| 18.1 | - |
starch | pH 6.0, 30°C, disproportionation activity, recombinant wild-type enzyme | Corynebacterium glutamicum | |
| 19.7 | - |
starch | pH 6.0, 30°C, disproportionation activity, recombinant mutant P228Y | Corynebacterium glutamicum | |
| 22.9 | - |
starch | pH 6.0, 30°C, disproportionation activity, recombinant mutant G417F | Corynebacterium glutamicum | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Corynebacterium glutamicum | Q8NNA7 | - |
- |
| Corynebacterium glutamicum 534 | Q8NNA7 | - |
- |
| Corynebacterium glutamicum ATCC 13032 | Q8NNA7 | - |
- |
| Corynebacterium glutamicum BCRC 11384 | Q8NNA7 | - |
- |
| Corynebacterium glutamicum CCUG 27702 | Q8NNA7 | - |
- |
| Corynebacterium glutamicum DSM 20300 | Q8NNA7 | - |
- |
| Corynebacterium glutamicum JCM 1318 | Q8NNA7 | - |
- |
| Corynebacterium glutamicum LMG 3730 | Q8NNA7 | - |
- |
| Corynebacterium glutamicum NBRC 12168 | Q8NNA7 | - |
- |
| Corynebacterium glutamicum NCIMB 10025 | Q8NNA7 | - |
- |
| Corynebacterium glutamicum NRRL B-2784 | Q8NNA7 | - |
- |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| additional information | the enzyme performs starch transglycosylation, starch degradation, disproportionation, hydrolysis, cyclization and coupling activities. The substrate specificity of wild-type and mutant enzymes for disproportionation activity is determined with malto-oligosaccharides (maltose (G2) to maltoheptaose (G7)), overview. The reaction products are analyzed by high-performance anion-exchange chromatography-pulsed amperometric detection (HPAEC-PAD) | Corynebacterium glutamicum | ? | - |
- |
|
| starch + maltoheptaose | - |
Corynebacterium glutamicum | ? | - |
? | |
| starch + maltohexaose | - |
Corynebacterium glutamicum | ? | - |
? | |
| starch + maltohexaose | - |
Corynebacterium glutamicum ATCC 13032 | ? | - |
? | |
| starch + maltohexaose | - |
Corynebacterium glutamicum DSM 20300 | ? | - |
? | |
| starch + maltohexaose | - |
Corynebacterium glutamicum JCM 1318 | ? | - |
? | |
| starch + maltohexaose | - |
Corynebacterium glutamicum LMG 3730 | ? | - |
? | |
| starch + maltohexaose | - |
Corynebacterium glutamicum NCIMB 10025 | ? | - |
? | |
| starch + maltohexaose | - |
Corynebacterium glutamicum BCRC 11384 | ? | - |
? | |
| starch + maltohexaose | - |
Corynebacterium glutamicum CCUG 27702 | ? | - |
? | |
| starch + maltohexaose | - |
Corynebacterium glutamicum NBRC 12168 | ? | - |
? | |
| starch + maltohexaose | - |
Corynebacterium glutamicum NRRL B-2784 | ? | - |
? | |
| starch + maltohexaose | - |
Corynebacterium glutamicum 534 | ? | - |
? | |
| starch + maltopentaose | - |
Corynebacterium glutamicum | ? | - |
? | |
| starch + maltopentaose | - |
Corynebacterium glutamicum ATCC 13032 | ? | - |
? | |
| starch + maltopentaose | - |
Corynebacterium glutamicum DSM 20300 | ? | - |
? | |
| starch + maltopentaose | - |
Corynebacterium glutamicum JCM 1318 | ? | - |
? | |
| starch + maltopentaose | - |
Corynebacterium glutamicum LMG 3730 | ? | - |
? | |
| starch + maltopentaose | - |
Corynebacterium glutamicum NCIMB 10025 | ? | - |
? | |
| starch + maltopentaose | - |
Corynebacterium glutamicum BCRC 11384 | ? | - |
? | |
| starch + maltopentaose | - |
Corynebacterium glutamicum CCUG 27702 | ? | - |
? | |
| starch + maltopentaose | - |
Corynebacterium glutamicum NBRC 12168 | ? | - |
? | |
| starch + maltopentaose | - |
Corynebacterium glutamicum NRRL B-2784 | ? | - |
? | |
| starch + maltopentaose | - |
Corynebacterium glutamicum 534 | ? | - |
? | |
| starch + maltose | - |
Corynebacterium glutamicum | ? | - |
? | |
| starch + maltose | - |
Corynebacterium glutamicum ATCC 13032 | ? | - |
? | |
| starch + maltose | - |
Corynebacterium glutamicum DSM 20300 | ? | - |
? | |
| starch + maltose | - |
Corynebacterium glutamicum JCM 1318 | ? | - |
? | |
| starch + maltose | - |
Corynebacterium glutamicum LMG 3730 | ? | - |
? | |
| starch + maltose | - |
Corynebacterium glutamicum NCIMB 10025 | ? | - |
? | |
| starch + maltose | - |
Corynebacterium glutamicum BCRC 11384 | ? | - |
? | |
| starch + maltose | - |
Corynebacterium glutamicum CCUG 27702 | ? | - |
? | |
| starch + maltose | - |
Corynebacterium glutamicum NBRC 12168 | ? | - |
? | |
| starch + maltose | - |
Corynebacterium glutamicum NRRL B-2784 | ? | - |
? | |
| starch + maltose | - |
Corynebacterium glutamicum 534 | ? | - |
? | |
| starch + maltotetraose | - |
Corynebacterium glutamicum | ? | - |
? | |
| starch + maltotetraose | - |
Corynebacterium glutamicum ATCC 13032 | ? | - |
? | |
| starch + maltotetraose | - |
Corynebacterium glutamicum DSM 20300 | ? | - |
? | |
| starch + maltotetraose | - |
Corynebacterium glutamicum JCM 1318 | ? | - |
? | |
| starch + maltotetraose | - |
Corynebacterium glutamicum LMG 3730 | ? | - |
? | |
| starch + maltotetraose | - |
Corynebacterium glutamicum NCIMB 10025 | ? | - |
? | |
| starch + maltotetraose | - |
Corynebacterium glutamicum BCRC 11384 | ? | - |
? | |
| starch + maltotetraose | - |
Corynebacterium glutamicum CCUG 27702 | ? | - |
? | |
| starch + maltotetraose | - |
Corynebacterium glutamicum NBRC 12168 | ? | - |
? | |
| starch + maltotetraose | - |
Corynebacterium glutamicum NRRL B-2784 | ? | - |
? | |
| starch + maltotetraose | - |
Corynebacterium glutamicum 534 | ? | - |
? | |
| starch + maltotriose | - |
Corynebacterium glutamicum | ? | - |
? | |
| starch + maltotriose | - |
Corynebacterium glutamicum ATCC 13032 | ? | - |
? | |
| starch + maltotriose | - |
Corynebacterium glutamicum DSM 20300 | ? | - |
? | |
| starch + maltotriose | - |
Corynebacterium glutamicum JCM 1318 | ? | - |
? | |
| starch + maltotriose | - |
Corynebacterium glutamicum LMG 3730 | ? | - |
? | |
| starch + maltotriose | - |
Corynebacterium glutamicum NCIMB 10025 | ? | - |
? | |
| starch + maltotriose | - |
Corynebacterium glutamicum BCRC 11384 | ? | - |
? | |
| starch + maltotriose | - |
Corynebacterium glutamicum CCUG 27702 | ? | - |
? | |
| starch + maltotriose | - |
Corynebacterium glutamicum NBRC 12168 | ? | - |
? | |
| starch + maltotriose | - |
Corynebacterium glutamicum NRRL B-2784 | ? | - |
? | |
| starch + maltotriose | - |
Corynebacterium glutamicum 534 | ? | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| amylomaltase | - |
Corynebacterium glutamicum |
| CgAM | - |
Corynebacterium glutamicum |
| GH77 amylomaltase | - |
Corynebacterium glutamicum |
| MalQ | - |
Corynebacterium glutamicum |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | - |
- |
Corynebacterium glutamicum |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.000755 | - |
starch | pH 6.0, 30°C, cyclization activity, recombinant mutant G417F | Corynebacterium glutamicum | |
| 0.00157 | - |
starch | pH 6.0, 30°C, cyclization activity, recombinant mutant Y23A | Corynebacterium glutamicum | |
| 0.00228 | - |
starch | pH 6.0, 30°C, cyclization activity, recombinant mutant P228Y | Corynebacterium glutamicum | |
| 0.00277 | - |
starch | pH 6.0, 30°C, cyclization activity, recombinant wild-type enzyme | Corynebacterium glutamicum | |
| 0.00279 | - |
starch | pH 6.0, 30°C, cyclization activity, recombinant mutant E231Y | Corynebacterium glutamicum | |
| 0.00545 | - |
starch | pH 6.0, 30°C, starch transglycosylation, recombinant mutant G417F | Corynebacterium glutamicum | |
| 0.00625 | - |
starch | pH 6.0, 30°C, starch transglycosylation, recombinant mutant Y23A | Corynebacterium glutamicum | |
| 0.0073 | - |
starch | pH 6.0, 30°C, starch transglycosylation, recombinant wild-type enzyme | Corynebacterium glutamicum | |
| 0.0073 | - |
starch | pH 6.0, 30°C, starch transglycosylation, recombinant mutant P228Y | Corynebacterium glutamicum | |
| 0.017 | - |
starch | pH 6.0, 30°C, starch transglycosylation, recombinant mutant E231Y | Corynebacterium glutamicum | |
| 43.83 | - |
starch | pH 6.0, 30°C, disproportionation activity, recombinant mutant P228Y | Corynebacterium glutamicum | |
| 103 | - |
starch | pH 6.0, 30°C, disproportionation activity, recombinant wild-type enzyme | Corynebacterium glutamicum | |
| 110.8 | - |
starch | pH 6.0, 30°C, disproportionation activity, recombinant mutant Y23A | Corynebacterium glutamicum | |
| 111.3 | - |
starch | pH 6.0, 30°C, disproportionation activity, recombinant mutant E231Y | Corynebacterium glutamicum | |
| 139.5 | - |
starch | pH 6.0, 30°C, disproportionation activity, recombinant mutant G417F | Corynebacterium glutamicum | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 6 | 6.5 | - |
Corynebacterium glutamicum |
General Information
| General Information | Comment | Organism |
|---|---|---|
| additional information | analysis of the functional amino acid positions of Corynebacterium glutamicum amylomaltase (CgAM) involved in LR-CD synthesis, overview. Molecular interactions analysis using the three-dimensional crystal structure of enzyme CgAM (PDB ID 5B68), computational analysis | Corynebacterium glutamicum |
| physiological function | enzyme amylomaltase has the ability to synthesize cyclic glucans, or large ring cyclodextrins (LR-CDs), with a degree of polymerization (DP) of more than eight through intramolecular transglycosylation reaction. The LR-CDs are highly soluble in water and relatively have a larger hydrophobic cavity compared to the small ring cyclodextrin | Corynebacterium glutamicum |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0448 | - |
starch | pH 6.0, 30°C, starch transglycosylation, recombinant mutant Y23A | Corynebacterium glutamicum | |
| 0.0773 | - |
starch | pH 6.0, 30°C, starch transglycosylation, recombinant mutant G417F | Corynebacterium glutamicum | |
| 0.0957 | - |
starch | pH 6.0, 30°C, starch transglycosylation, recombinant wild-type enzyme | Corynebacterium glutamicum | |
| 0.0957 | - |
starch | pH 6.0, 30°C, starch transglycosylation, recombinant mutant E231Y | Corynebacterium glutamicum | |
| 0.099 | - |
starch | pH 6.0, 30°C, starch transglycosylation, recombinant mutant P228Y | Corynebacterium glutamicum | |
| 2.23 | - |
starch | pH 6.0, 30°C, disproportionation activity, recombinant mutant P228Y | Corynebacterium glutamicum | |
| 5.7 | - |
starch | pH 6.0, 30°C, disproportionation activity, recombinant wild-type enzyme | Corynebacterium glutamicum | |
| 6.09 | - |
starch | pH 6.0, 30°C, disproportionation activity, recombinant mutant G417F | Corynebacterium glutamicum | |
| 7.23 | - |
starch | pH 6.0, 30°C, disproportionation activity, recombinant mutant E231Y | Corynebacterium glutamicum | |
| 9.03 | - |
starch | pH 6.0, 30°C, disproportionation activity, recombinant mutant Y23A | Corynebacterium glutamicum | |
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