Enzymes are biocatalysts with complex structures and specific catalytic mechanisms that determine their distinctive properties, such as high catalytic activity and selectivity of specific substrates. Oxidoreductase (OXR) enzymes are in high demand for biocatalytic applications in the food industry and cosmetics (glucose oxidase (GOx) and cellobiose dehydrogenase (CDH)), bioremediations (horseradish peroxidase (HRP) and laccase (LAC)), and medicine for biosensors and miniature biofuel cells (GOx, CDH, LAC, and HRP). Therefore, scientists are still trying to find optimal fermentation formulas and, most recently, also using protein engineering and directed evolution for an additional increase in the yield of recombinant enzyme production.
Source | Variant of Oxidoreductase Enzyme | Host Strain; Vector | Promoter | Inducer | Signal Sequence | Additional Information | Enzyme Yield | Ref. |
---|---|---|---|---|---|---|---|---|
GOx | ||||||||
A. niger | NR | 2805; YEp352 | GAL1 | 1% galactose | ss of α-factor | NR | 32 a U/mL | [36] |
A. oryzae | NR | 2805 | GAL-10 | NR | α-amylase signal sequence | 30 °C, 150 rpm, feedback-controlled fed-batch | NR | [37] |
A. niger | NR | 2805; Yep352 | Hybrid ADH2-GPD | 2% glucose | ss of α-factor | 1.5% EtOH | 260 a U/mL | [36] |
A. niger | NR | GRF181 pSGO2 |
ADH2-GPD | 8% sucrose | Native | Shake flask; 28 °C; 200 h | 106 a U/mL | [38] |
CDH | ||||||||
M. thermophilum | Wild type | BJ5465; pJRoC30 | GAL1 | 2% galactose | Native | Deep-well plate (500 µL of medium); 30 °C; 5 days | 50 b U/L | [39] |
M. thermophilum | Wild type | BJ5465; pJRoC30 | GAL1 | 2% galactose | ss of α factor | Deep-well plate (500 µL of medium); 30 °C; 5 days | 16 b U/L | [39] |
P. chrysosporium | U46081.1 | InvSC1; pYES2 | GAL1 | Galactose | Native | Shake flask; 30 °C; 16 h | NR | [40] |
T. clypeatus | GAFV01008428.1 | BY4742; pFL61 | PGK | No | No | NR type of cultivation; Czapek medium, 3 days | 0.039 b U/mg | [41] |
HRP | ||||||||
Horseradish | Wild type | SIP-Ost1 (Δ44–70); modified pESC-URA | TDH3 | pre-Ost1 | Fermenter 5 L (batch fermentation) | 13,506 c U/L | [42] | |
Horseradish | HRP 3-17E12 | BJ5465; pYEX-S1 | PGK1 | No | NR | Expression time 25 h | about 250 c U/L | [43] |
LAC | ||||||||
M. thermophila | MtL | BJ5465; pJRoC3 | NR | NR | NR | Shake flask 2.8 L; 0.005 mM CuSO4 30 °C; 1 day | 0.6 d U/L | [44] |
M. thermophila | T2 mutant | BJ5465; pJRoC3 | NR | NR | NR | Shake flask 2.8 L; 0.005 mM CuSO4; 30 °C; 1 day | 102 d U/L | [44] |
T. versicolor | Cvl3 | BY2777; pYES2 | GAL1 | 4% Galactose | Native | Shake flask 0.3 L; 0.5 mM CuSO4; 20 °C; 6 days | 45 e U/L | [45] |
L. edodes | Lcc4 | FGY217; pBG13 | GAL1 | 4% Galactose | Native | Fermentor 4 L; 0.5 mM CuSO4; 20 °C; 7 days | 10 e U/L | [46] |
A. pediades | ApL | BJ5465; pJRoC30 | GAL1 | 2.2% Galactose | α9H2 signal peptide | Shake flask 0.1 L; 0.4 mM CuSO4; 20 °C; 4 days | 280 e U/L | [47] |
Trametes sp. C30 | Clac1, 2, 3 | W303-1A; YIp351 | PGK1 | No | ss of SUC2 gene product | Fermentor 3 L; 1 mM CuSO4; 28 °C; 3 days | 1200 d U/L | [48] |
M. thermophila | T2 mutant | BW31a; pVT-100U | ADH1 | No | Native | Shake flask 0.25 L; 0.6 mM CuSO4; 30 °C; 1 day; 0.8% alanine | 6.52 e U/L | [49] |
T. versicolor | Lcc1 | BW31a; pVT-100U | ADH1 | No | Native | Shake flask 0.25 L; 0.6 mM CuSO4; 30 °C; 1 day; 0.8% alanine | 0.45 e U/L | [49] |
T. trogii | Lcc1 | BW31a; pVT-100U | ADH1 | No | Native | Shake flask 0.25 L; 0.6 mM CuSO4; 20 °C; 14 days; 0.8% alanine | 14.12 e U/L | [49] |
Source | Variant of Oxidoreductase Enzyme | Host Strain; Vector | Promoter | Inducer | Signal Sequence | Additional Information | Enzyme Yield | Ref. |
---|---|---|---|---|---|---|---|---|
GOx | ||||||||
A. niger | GOx accc30161 | SMD1168;pGAPZαA | GAP | NR | ss of α-factor | 30 °C; pH 6 | 107.18 a U/mL | [59] |
A. niger | GOxM | SMD1168; pPIC3.5 | AOX1 | 1% MeOH | 30 °C; 3 days, 220 rpm | 26.93 a U/mL | [60] | |
A. niger ATCC 9029 | - | GS115; pPIC9 | AOX1 | 1% MeOH | 28 °C; 225 rpm | NR | [61] | |
A. niger | M12 mutant | KM71H; pPICZαA | AOX | 0.5% MeOH | Proalpha sequence | Nine days of fermentation | 17.5 b U/mL | [62] |
CDH | ||||||||
M. thermophilum | N700S mutant | X33; pPICZαA | AOX1 | 0.5% MeOH | ss of α-factor & propeptide | Fermentor 7 L; 30 °C; 5 days | 1800 c U/L | [39] |
P. cinnabarinus | Wild type | X33; pPICZαA | AOX1 | 3% MeOH | ss of α-factor | Fermentor 1 L; 4 days | 7800 c U/L | [63] |
N. crassa strain FGSC 2489 | NC-cdh1 | X33; pPICZαB | AOX1 | 1% MeOH | ss of α-factor | Shake flask 0.25 L; 30 °C; 1 day | 7451 c U/L | [64] |
P. chrysosporium | Mutant | KM71H; pPICZαA | AOX1 | 0.5% MeOH | ss of α-factor | Shake flask; 28 °C; 6 days | 950 c U/L | [64] |
HRP | ||||||||
Horseradish | wild type | X-33; pPICZαB | AOX1 | 0.5% MeOH | ss of α-factor | 30 °C; BMGY medium supplemented with 1% casamino acids; BMMY medium supplemented with 1.0 mM vitamin B1, 1.0 mM δ-ALA, and trace element mix; the highest yield in 80–90 h post-induction | 377 d U/mg | [43] |
Horseradish | mutant HRP 2-13A10 | X-33; pPICZαB | AOX1 | 0.5% MeOH | ss of α-factor | Same as for wild-type | 2053 d U/mg | [43] |
Horseradish | mutant HRP 3-17E12 | X-33; pPICZαB | AOX1 | 0.5% MeOH | ss of α-factor | Same as for wild-type | 1049 d U/mg | [43] |
Horseradish | A2A isoenzyme | X-33; pPICZαC | AOX1 | 0.5% MeOH | α-MF-pre-pro signal peptide | BMMY medium supplemented with 1% casamino acids and 1% sorbitol | 25.63 a U/mg | [65] |
Horseradish | HRP-SpG | PpFWK3; pPpT4_alpha_S | AOX1 | MeOH | NR | 136 h of methanol induction | 113 d mg/L | [66] |
LAC | ||||||||
C. cinerea | Lcc9 | X33; pGAPZαA | GAP | 0.5% glucose | ss of α-factor | Shake flasks 0.25 L; 0.3 mM CuSO4; 20 °C; 4 days; 0.8% alanine | 12.8 f µkat/L | [67] |
P. ostreatus | rPOXA 1B | X33; pGAPZαA | GAP | 0.5% glucose | ss of α-factor | Bioreactor 10 L; 1 mM CuSO4; 2% peptone; 1.5% yeast extract; 170 h; geometry of flask | 3159.93 f U/L | [68] |
T. versicolor | Lcc1 | SMD 1168; pHIL-D2 | AOX1 | 0.5% MeOH | Shake flasks 1 L; 0.1 mM CuSO4; 20 °C; 3 days of induction | 11,500 f U/L | [69] | |
T. versicolor | Lcc1 | SMD 1168; pHIL-D2 | AOX1 | 0.5% MeOH | BioFlo III fermentor; 0.1 mM CuSO4; 20 °C; 8.5 days | 140 f U/L | [69] | |
T. versicolor | Lcc1 | GS115; pPIC3.5 | AOX1 | 1% MeOH | Shake flasks (0.1 L; 0.2 mM CuSO4; 22 °C; initial pH 6; 0.8% alanine: | 23.9 f U/L | [70] | |
T. versicolor | LccA | X33; pPICZαB | AOX | 0.6% MeOH | ss of α-factor | Shake flask (0.05 L of medium); 0.5 mM CuSO4; 28 °C; 16 days; initial pH 7; | 11.972 f U/L | [71] |
T. versicolor | LccA | X33; X33; pPICZαB | AOX | 0.6% MeOH | ss of α-factor | 5 L fermenter; 0.5 mM CuSO4; 28 °C; 4.2 days; initial pH 7; | 18.123 f U/L | [71] |
C. gallica | LcCg | X33; pPICZB | AOX | 1% MeOH | Modified α-factor preproleader | Fernbach flask; 0.5 mM CuSO4; 28 °C; 12 days; initial pH 6; 0.8% alanine | 250 e U/L | [72] |
Trameters sp. 48424 | Lac48424-1 | GS115; pPIC3.5K | AOX | 0.5% MeOH | Native | Shake flasks; 0.3 mM CuSO4; 20 °C; 7 days; initial pH 6; 0.8% alanine | 104.45 f U/L | [73] |
C. cinerea | Lcc9 | GS115; pPIC9K | AOX | 0.5% MeOH | Native | Shake flasks 0.5 L; 0.3 mM CuSO4; 28 °C; 10 days; initial pH 6.5; 0.8% alanine | 3138 ± 62 f U/L | [74] |
C. cinerea | Lcc9 | X33; pPICZαA | AOX | 0.5% MeOH | ss of α-factor | Shake flasks 0.25 L; 0.3 mM CuSO4; 20 °C; 7 days; 0.8% alanine | 9.3 f µkat/L | [67] |