Cloned (Comment) | Organism |
---|---|
gene mbdh, functional coexpression of Serratia sp. meso-2,3-butanediol dehydrogenase, Lactobacillus brevis NADH oxidase and Vitreoscilla sp. hemoglobin in Escherichia coli strain BL21(DE3)/pET-mbdh-nox-vgb, construction of plasmids pET-mbdh, pET-mbdh-nox, and pET-mbdh-nox-vgb, method optimization | Serratia sp. |
gene mbdh, recombinant expression of meso-2,3-BDH in Escherichia coli strain BL21(DE3), coexpression with NADH oxidase (NOX) from Lactobacillus brevis, and hemoglobin protein (VHB) from Vitreoscilla, subcloning in Escherichia coli strain DH5alpha | Serratia sp. T241 |
Protein Variants | Comment | Organism |
---|---|---|
additional information | efficient (3R)-acetoin production from meso-2,3-butanediol using a whole-cell biocatalyst with coexpression of Serratia sp. meso-2,3-butanediol dehydrogenase, Lactobacillus brevis NADH oxidase and Vitreoscilla sp. hemoglobin | Serratia sp. |
additional information | chiral (3R)-AC production from meso-2,3-butanediol (meso-2,3-BD) is obtained using recombinant Escherichia coli cells co-expressing meso-2,3-butanediol dehydrogenase (meso-2,3-BDH), NADH oxidase (NOX), and hemoglobin protein (VHB) from Serratia sp. T241, Lactobacillus brevis, and Vitreoscilla, respectively. The biocatalysis system of Escherichia coli/pET-mbdh-nox-vgb is developed and the bioconversion conditions are optimized. Under the optimal conditions, 86.74 g/l of (3R)-acetoin with the productivity of 3.61 g/l/h and the stereoisomeric purity of 97.89% is achieved from 93.73 g/l meso-2,3-BD using the whole-cell biocatalysis system, pH 7.0 at 30°C for 12 h. The results show the industrial potential for (3R)-acetoin production via whole-cell biocatalysis. Escherichia coli/pET-mbdh cannot produce acetoin from (2R,3R)-2,3-BD as substrate. To obtain high (3R)-acetoin productivity, a cofactor regeneration system involved in co-expression of meso-2,3-BDH and NOX enzymes from Serratia sp. T241 Lactobacillus brevis is developed in Escherichia coli. The NOX enzyme efficiently oxidizes NADH, which is formed by meso-2,3-BDH, and regenerate NAD+ for the biocatalytic process. The feasibility of (3R)-AC production from the substrate of meso-2,3-BD by whole-cell biocatalysis is conducted, method optimization, overview. A small amount of (3S)-acetoin (1.86 g/l) can also be produced from (2S,3S)-2,3-BD in the substrate 2,3-BD (2.23% of (2S,3S)-2,3-BD) by the biocatalyst | Serratia sp. T241 |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | activates | Serratia sp. T241 | |
Mg2+ | activates | Serratia sp. T241 | |
additional information | poor effects by Mn2+, Fe3+, and Ca2+ | Serratia sp. T241 |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(2R,3S)-butane-2,3-diol + NAD+ | Serratia sp. | - |
(3R)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | Serratia sp. T241 | - |
(3R)-acetoin + NADH + H+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Serratia sp. | AEF50077 | - |
- |
Serratia sp. T241 | - |
- |
- |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
2.17 | - |
crude extracet of Escherichia coli cells expressing the enzyme from plasmid pET-mbdh-nox-vgb, pH 8.0, 22°C | Serratia sp. |
2.47 | - |
crude extracet of Escherichia coli cells expressing the enzyme from plasmid pET-mbdh-nox, pH 8.0, 22°C | Serratia sp. |
3.32 | - |
crude extracet of Escherichia coli cells expressing the enzyme from plasmid pET-mbdh, pH 8.0, 22°C | Serratia sp. |
3.32 | - |
recombinant enzyme, pH 8.0, 22°C | Serratia sp. T241 |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(2R,3S)-butane-2,3-diol + NAD+ | - |
Serratia sp. | (3R)-acetoin + NADH + H+ | - |
r | |
(2R,3S)-butane-2,3-diol + NAD+ | - |
Serratia sp. T241 | (3R)-acetoin + NADH + H+ | - |
? | |
additional information | meso-2,3-BDH from Serratia sp. T241 exhibits higher catalytic efficiency compared with the meso-2,3-BDHs from Klebsiella pneumoniae strain XJ-Li and Serratia marcescens strain H30. No activity is detected for (2R,3R)-2,3-BD as substrate by meso-2,3-BDH, but meso-2,3-BDH from Serratia sp. T241 can efficiently convert (2S,3S)-2,3-BD and meso-2,3-BD into (3S)-acetoin and (3R)-acetoin, respectively, cf. EC 1.1.1.76 | Serratia sp. T241 | ? | - |
- |
Synonyms | Comment | Organism |
---|---|---|
mbdh | - |
Serratia sp. |
mbdh | - |
Serratia sp. T241 |
meso-2,3-BDH | - |
Serratia sp. |
meso-2,3-BDH | - |
Serratia sp. T241 |
meso-2,3-butanediol dehydrogenase | - |
Serratia sp. |
More | cf. EC 1.1.1.76 | Serratia sp. T241 |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
22 | - |
assay at room temperature | Serratia sp. T241 |
22 | - |
in vitro assay at room temperature | Serratia sp. |
30 | 37 | in vivo assay at | Serratia sp. |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
in vivo assay at | Serratia sp. |
8 | - |
assay at | Serratia sp. T241 |
8 | - |
in vitro assay at | Serratia sp. |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Serratia sp. | |
NAD+ | - |
Serratia sp. T241 | |
NADH | - |
Serratia sp. |