Activating Compound | Comment | Organism | Structure |
---|---|---|---|
additional information | no effect by 1 mM DTT on wild-type enzyme activity | Talaromyces purpureogenus | |
SDS | activates the wild-type enzyme by 62% at 0.1% v/v | Talaromyces purpureogenus | |
Triton X-100 | activates the wild-type enzyme by 33% at 0.1% v/v | Talaromyces purpureogenus |
Application | Comment | Organism |
---|---|---|
synthesis | endoinulinases are an industrial tool critical for the production of inulooligosaccharides (IOS) | Talaromyces purpureogenus |
Cloned (Comment) | Organism |
---|---|
DNA and amino acid sequence determination and analysis, recombinant expression of MBP-tagged enzyme in Escherichia coli strain BL21 | Talaromyces purpureogenus |
Protein Variants | Comment | Organism |
---|---|---|
additional information | directed evolution yields variants showing up to 5fold improvements in soluble enzyme production compared to the starting point which enables high-yield production of highly purified recombinant enzyme. The distribution of the enzymatic reaction products demonstrates that after 24 h of incubation, the main product (57%) has a degree of polymerization of 3 (DP3). The MBP-fused wild-type enzyme shows increased activity compared to unaltered wild-type enzyme. The evolved endoinulinase mutant variants exhibit increased solubility and activity compared to wild-type | Talaromyces purpureogenus |
Y128H | site-directed mutagenesis, the mutation Y128H is in close proximity to the active site, it leads to modified enzyme catalytic activity with increased kcat and kcat/KM compared to untagged wild-type enzyme, while the activity of MBP-tagged wild-type enzyme is higher | Talaromyces purpureogenus |
Y128H/A316T/E344K/T504M | site-directed mutagenesis, the mutant shows reduced activity compared to wild-type enzyme and MBP-fused wild-type enzyme | Talaromyces purpureogenus |
Y128H/E344K/T504M | site-directed mutagenesis, the mutation E344K is located on the enzyme protein surface, mutation Y128H is in close proximity to the active site. The mutant shows reduced activity compared to wild-type enzyme and MBP-fused wild-type enzyme | Talaromyces purpureogenus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Al3+ | 64% inhibition of wild-type enzyme at 1 mM | Talaromyces purpureogenus | |
Cu2+ | 50% inhibition of wild-type enzyme at 1 mM | Talaromyces purpureogenus | |
EDTA | 68% inhibition of wild-type enzyme at 1 mM | Talaromyces purpureogenus | |
additional information | no effect by 1 mM DTT on enzyme activity | Talaromyces purpureogenus | |
Ni2+ | 14% inhibition of wild-type enzyme at 1 mM | Talaromyces purpureogenus | |
Zn2+ | 34% inhibition of wild-type enzyme at 1 mM | Talaromyces purpureogenus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.21 | - |
inulin | pH 5.4, 55°C, recombinant untagged wild-type enzyme | Talaromyces purpureogenus | |
0.78 | - |
inulin | pH 5.4, 55°C, recombinant MBP-tagged wild-type enzyme | Talaromyces purpureogenus | |
2.4 | - |
inulin | pH 5.4, 55°C, recombinant MBP-tagged mutant Y128H/E344K/T504M enzyme | Talaromyces purpureogenus | |
2.82 | - |
inulin | pH 5.4, 55°C, recombinant MBP-tagged mutant Y128H enzyme | Talaromyces purpureogenus | |
3.12 | - |
inulin | pH 5.4, 55°C, recombinant MBP-tagged mutant Y128H/A316T/E344K/T504M enzyme | Talaromyces purpureogenus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | activates the wild-type enzyme 16% at 1 mM | Talaromyces purpureogenus | |
Mg2+ | activates the wild-type enzyme 32% at 1 mM | Talaromyces purpureogenus | |
additional information | no effect by 1 mM K+ on enzyme activity | Talaromyces purpureogenus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Talaromyces purpureogenus | O00056 | i.e. Penicillium purpureogenum | - |
Talaromyces purpureogenus ATCC 4713 | O00056 | i.e. Penicillium purpureogenum | - |
Purification (Comment) | Organism |
---|---|
recombinant MBP-tagged enzyme from Escherichia coli strain BL21 by amylose affinity chromatography, and gel filtration | Talaromyces purpureogenus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
inulin + H2O | - |
Talaromyces purpureogenus | ? | - |
? | |
inulin + H2O | - |
Talaromyces purpureogenus ATCC 4713 | ? | - |
? | |
additional information | the enzyme targets the internal beta-2,1 fructofuranosidic linkages of inulin to yield both GFn and Fm molecules (G for glucose, F for fructose, n and m indicate the number of fructose moieties) | Talaromyces purpureogenus | ? | - |
? | |
additional information | the enzyme targets the internal beta-2,1 fructofuranosidic linkages of inulin to yield both GFn and Fm molecules (G for glucose, F for fructose, n and m indicate the number of fructose moieties) | Talaromyces purpureogenus ATCC 4713 | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
beta-2,1-D-fructan fructanohydrolase | - |
Talaromyces purpureogenus |
endo-inulinase | - |
Talaromyces purpureogenus |
endoinulinase | - |
Talaromyces purpureogenus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
55 | - |
recombinant wild-type and mutant enzymes | Talaromyces purpureogenus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
120 | - |
inulin | pH 5.4, 55°C, recombinant untagged wild-type enzyme | Talaromyces purpureogenus | |
685 | - |
inulin | pH 5.4, 55°C, recombinant MBP-tagged mutant Y128H/A316T/E344K/T504M enzyme | Talaromyces purpureogenus | |
792 | - |
inulin | pH 5.4, 55°C, recombinant MBP-tagged mutant Y128H/E344K/T504M enzyme | Talaromyces purpureogenus | |
850 | - |
inulin | pH 5.4, 55°C, recombinant MBP-tagged wild-type enzyme | Talaromyces purpureogenus | |
1797 | - |
inulin | pH 5.4, 55°C, recombinant MBP-tagged mutant Y128H enzyme | Talaromyces purpureogenus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5.4 | - |
assay at | Talaromyces purpureogenus |
General Information | Comment | Organism |
---|---|---|
additional information | enzyme structure homology modeling using the structure of endoinulinase from Aspergillus ficcum (PDB ID 3RWK) as template | Talaromyces purpureogenus |
physiological function | inulinases are fructofuranosyl hydrolases that target the beta-2,1 linkage of inulin and hydrolyze it into fructose, glucose and inulooligosaccharides (IOS), IOS are functioning as dietary fibers | Talaromyces purpureogenus |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
219.55 | - |
inulin | pH 5.4, 55°C, recombinant MBP-tagged mutant Y128H/A316T/E344K/T504M enzyme | Talaromyces purpureogenus | |
330 | - |
inulin | pH 5.4, 55°C, recombinant MBP-tagged mutant Y128H/E344K/T504M enzyme | Talaromyces purpureogenus | |
571.43 | - |
inulin | pH 5.4, 55°C, recombinant untagged wild-type enzyme | Talaromyces purpureogenus | |
637.23 | - |
inulin | pH 5.4, 55°C, recombinant MBP-tagged mutant Y128H enzyme | Talaromyces purpureogenus | |
1089.74 | - |
inulin | pH 5.4, 55°C, recombinant MBP-tagged wild-type enzyme | Talaromyces purpureogenus |