Protein Variants | Comment | Organism |
---|---|---|
additional information | construction of multienzyme supramolecular complexes, which self-assemble into spatially defined architectures, to improve the efficiency of cascade reactions. Engineered supramolecular enzyme assemblies enhance hexose 6-phosphate and fructose 6-phosphate production and can be similarly created as a kinetic trap to enable fast and efficient methanol utilization, method, overview. Clustering Mdh3 with a bifunctional Hps-Phi fusion, further improves fructose 6-phosphate production, resulting in an overall 50fold improvement over the uncomplexed enzyme mixture. Compared with the unassembled enzyme system, a much lower level of formaldehyde is detected and only a small amount of hexose 6-phosphate is accumulated, indicating the effective channeling of formaldehyde toward fructose 6-phosphate by the supramolecular enzyme complex due to improved molecular proximity | Bacillus methanolicus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
methanol + NAD+ | Bacillus methanolicus | in nature, methanol dehydrogenase (Mdh), which converts methanol to formaldehyde, highly favors the reverse reaction | formaldehyde + NADH + H+ | - |
r | |
methanol + NAD+ | Bacillus methanolicus MGA3 | in nature, methanol dehydrogenase (Mdh), which converts methanol to formaldehyde, highly favors the reverse reaction | formaldehyde + NADH + H+ | - |
r | |
additional information | Bacillus methanolicus | efficient coupling with the irreversible sequestration of formaldehyde by 3-hexulose-6-phosphate synthase (Hps) and 6-phospho-3-hexuloseisomerase (Phi) serves as the key driving force to pull the pathway equilibrium toward central metabolism | ? | - |
? | |
additional information | Bacillus methanolicus MGA3 | efficient coupling with the irreversible sequestration of formaldehyde by 3-hexulose-6-phosphate synthase (Hps) and 6-phospho-3-hexuloseisomerase (Phi) serves as the key driving force to pull the pathway equilibrium toward central metabolism | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus methanolicus | I3E949 | - |
- |
Bacillus methanolicus MGA3 | I3E949 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
methanol + NAD+ | - |
Bacillus methanolicus | formaldehyde + NADH + H+ | - |
r | |
methanol + NAD+ | in nature, methanol dehydrogenase (Mdh), which converts methanol to formaldehyde, highly favors the reverse reaction | Bacillus methanolicus | formaldehyde + NADH + H+ | - |
r | |
methanol + NAD+ | - |
Bacillus methanolicus MGA3 | formaldehyde + NADH + H+ | - |
r | |
methanol + NAD+ | in nature, methanol dehydrogenase (Mdh), which converts methanol to formaldehyde, highly favors the reverse reaction | Bacillus methanolicus MGA3 | formaldehyde + NADH + H+ | - |
r | |
additional information | efficient coupling with the irreversible sequestration of formaldehyde by 3-hexulose-6-phosphate synthase (Hps) and 6-phospho-3-hexuloseisomerase (Phi) serves as the key driving force to pull the pathway equilibrium toward central metabolism | Bacillus methanolicus | ? | - |
? | |
additional information | efficient coupling with the irreversible sequestration of formaldehyde by 3-hexulose-6-phosphate synthase (Hps) and 6-phospho-3-hexuloseisomerase (Phi) serves as the key driving force to pull the pathway equilibrium toward central metabolism | Bacillus methanolicus MGA3 | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
MDH | - |
Bacillus methanolicus |
Mdh3 | - |
Bacillus methanolicus |
NAD-dependent methanol dehydrogenase | - |
Bacillus methanolicus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Bacillus methanolicus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.4 | - |
assay at | Bacillus methanolicus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Bacillus methanolicus | |
NADH | - |
Bacillus methanolicus |
General Information | Comment | Organism |
---|---|---|
additional information | in nature, methanol dehydrogenase (Mdh), which converts methanol to formaldehyde, highly favors the reverse reaction, efficient coupling with the irreversible sequestration of formaldehyde by 3-hexulose-6-phosphate synthase (Hps) and 6-phospho-3-hexuloseisomerase (Phi) serves as the key driving force to pull the pathway equilibrium toward central metabolism. An emerging strategy to promote efficient substrate channeling is to spatially organize pathway enzymes in an engineered assembly to provide kinetic driving forces that promote carbon flux in a desirable direction. A scaffoldless, self-assembly strategy is applied to organize Mdh, Hps, and Phi into an engineered supramolecular enzyme complex using an SH3-ligand interaction pair, which enhances methanol conversion to fructose-6-phosphate. An NADH sink is created using Escherichia coli lactate dehydrogenase as an NADH scavenger, thereby preventing reversible formaldehyde reduction, to increase methanol consumption. Combination of the two strategies improves in vitro fructose 6-phosphate production by 97fold compared with unassembled enzymes. The beneficial effect of supramolecular enzyme assembly is also realized in vivo as the engineered enzyme assembly improves whole-cell methanol consumption rate by ninefold. This approach ultimately allows direct coupling of enhanced fructose 6-phosphate synthesis with other metabolic engineering strategies for the production of many desired metabolites from methanol | Bacillus methanolicus |