Table 1.
Characterization of the different AAO variants.
| Variant | Evolutionary campaign | Mutations | Library creation method | Heterologous host | Secretion level | Total activity improvement(in fold) | T50 (°C) | Specific activity | Catalytic efficiency(kcat/Km) | Reference |
|---|---|---|---|---|---|---|---|---|---|---|
| preαproK-AAO(parental type) | Functional expression | --- | --- | S. cerevisiae | N.D1 | 1 | 58.8 | N.D1 | N.D1 | [51] |
| FX7 | Functional expression | H91N | MORPHING | S. cerevisiae P. pastoris | 2 mg/L (S.c)N.D. (P.p) | 962 | 64.3 | 24 U/mg3 | 15623 | [51] |
| FX9 | Functional expression | F[3α]S N[25K]D T[50K]A F[52K]L L170M H91N | DNA Shuffling MORPHING | S. cerevisiae P. pastoris | 4.5 mg/L (S.c)4.3 mg/L (P.p) | 3502 | 63 | 34 U/mg3 | 17823 | [53] |
| LanDo | Secondary alcohols oxidation | F[3α]S N[25K]D T[50K]A F[52K]L L170M A77V R80C H91N V340A I500M F501W | CSM MORPHING Ep-PCR SDR | S. cerevisiae | 4.6 mg/L | 8004 | N.D | 2.9 U/mg5 | 7.55 | [54] |
| Bantha | Furfural derivatives transformation | F[3α]S N[25K]D T[50K]A F[52K]L L170M H91N F501W | CSM | S. cerevisiae | N.D. | 36 | N.D | N.D | 0.767 | [55] |
The first parental type was not purified to homogeneity since there was not enough protein functionally expressed. 2 Calculated with reference to parental preαproK-AAO for the oxidation of p-methoxybenzyl alcohol. 3 With p-methoxybenzyl alcohol as substrate. 4 Calculated with reference to parental FX9 for the oxidation of 1-(p-methoxyphenyl)-ethanol. 5 With 1-(p-methoxyphenyl)-ethanol as substrate. 6 Calculated with reference to parental FX9 for the oxidation of 5-hydroxymethyl furfural. 7 With 5-hydroxymethyl furfural as substrate. N.D: No determined. S.c: Saccharomyces cerevisiae. P.p: Pichia pastoris. CSM: Combinatorial Saturation Mutagenesis. Ep-PCR: Error prone PCR. SDR: Site-Directed Recombination.