Table 2.
Biosynthesis of pentacyclic triterpenoids using engineered yeasts.
| Parent Strain | Modification | Product (Yield) | Reference |
|---|---|---|---|
| Saccharomyces cerevisiae INVSc1 | Insertion of βAS, CPR (Lotus japonicus), CYP93E2, and CYP72A61 (Medicago truncatula) | β-Amyrin (6 **, 1.07 mg/L), 24-hydroxy-β-amyrin (13, 0.27 mg/L), and soyasapogenol B (21, 1.35 mg/L) | [67] |
| Insertion of βAS, CPR (L. japonicus), CYP716A12, and CYP72A68 (M. truncatula) | β-Amyrin (6, 0.55 mg/L), erythrodiol (8, 0.09 mg/L), and gypsogenic acid (22, 0.96 mg/L) | ||
| Insertion of βAS, CPR (L. japonicus), CYP716A12, and CYP93E2 (M. truncatula) | Erythrodiol (8), OA, 4-epi-hederagenin (24), and others (trace amount) | ||
| Insertion of βAS, CPR (L. japonicus), CYP72A63, and CYP716A12 (M. truncatula) | Erythrodiol (8), OA, queretaroic acid (25), and others (trace amount) | ||
| Insertion of βAS, CPR (L. japonicus), CYP93E2, and CYP72A63 (M. truncatula) | Probable 11-deoxo-GA (23) and others (trace amount) | ||
| S. cerevisiae BY4742 | Insertion of βAS (Glycyrrhiza glabra) | β-Amyrin (6, 36.2 ± 3.9 mg/L) | [71] |
| Insertion of βAS (M. truncatula) | β-Amyrin (6, 19.0 ± 1.0 mg/L) | ||
| S. cerevisiae TM3 | Insertion of AtATR1 (Arabidopsis thaliana), CYP716Y1 (Bupleurum falcatum), and CYP716A12 (M. truncatula) | β-Amyrin (6), erythrodiol (8), OA (1), oleanolic aldehyde (9), and 16α-hydroxy-oleanolic aldehyde (10) | |
| Insertion of CYP716Y1 (B. falcatum), CYP716A12 (M. truncatula), AtATR1 (A. thaliana), and UGT73C11 (Barbarea vulgaris) | 3-O-Glc-Echinocystic acid (11), 3-O-Glc-OA (12) | ||
| Insertion of AtATR1 (A. thaliana) and CYP93E2 (M. truncatula) | 24-Hydroxy-β-amyrin (13, 1.3%) | [72] | |
| Insertion of AtATR1 (A. thaliana) and CYP93E7 (Lens culinaris) | 24-Hydroxy-β-amyrin (13, 16.2%) | ||
| Insertion of AtATR1 (A. thaliana) and CYP93E8 (Pisum sativum) | 24-Hydroxy-β-amyrin (13, 37.6%), probable 3β-hydroxy-olean-12-en-24-oic acid (14, 3.2%) | ||
| Insertion of AtATR1 (A. thaliana) and CYP93E5 (Cicer arietinum) | 24-Hydroxy-β-amyrin (13, 51.3%), probable 3β-hydroxy-olean-12-en-24-oic acid (14, 7.6%) | ||
| Insertion of AtATR1 (A. thaliana) and CYP93E6 (G. glabra) | 24-Hydroxy-β-amyrin (13, 50.2%), probable 3β-hydroxy-olean-12-en-24-oic acid (14, 6.3%) | ||
| Insertion of AtATR1 (A. thaliana) and CYP93E4 (Arachis hypogaea) | 24-Hydroxy-β-amyrin (13, 47.0%), probable 3β-hydroxy-olean-12-en-24-oic acid (14, 5.5%) | ||
| Insertion of AtATR1 (A. thaliana) and CYP93E1 (Glycine max) | 24-Hydroxy-β-amyrin (13, 58.8%), probable 3β-hydroxy-olean-12-en-24-oic acid (14, 13.5%) | ||
| Insertion of AtATR1 (A. thaliana) and CYP93E9 (Phaseolus vulgaris) | 24-Hydroxy-β-amyrin (13, 67.6%), probable 3β-hydroxy-olean-12-en-24-oic acid (14, 11.8%) | ||
| S. cerevisiae BY4742 | Deletion of TRP1 | Squalene (4, 9.6 mg/L) | [66] |
| S. cerevisiae BY4742-TRP | Overexpression of tHMG1 and LYS2 | Squalene (4, 150.9 mg/L) | |
| S. cerevisiae BY-T1 | Insertion of βAS (G. glabra) Overexpression of ERG1 and ERG9 |
β-Amyrin (6, 107.0 mg/L), squalene (4, 183.4 mg/L) | |
| Insertion of βAS (Panax ginseng) Overexpression of ERG1 and ERG9 |
β-Amyrin (6, 1.9 mg/L) | ||
| S. cerevisiae BY-βA-G | Insertion of βAS (G. glabra), OAS (M. truncatula), and AtCPR1 (A. thaliana) | OA (1, 71.0 mg/L), β-amyrin (6, 88.6 mg/L), and squalene (4, 141.2 mg/L) | |
| S. cerevisiae INVSc1 | Insertion of βAS (G. glabra) | β-Amyrin (6, 4.16 mg/L) | [73] |
| Insertion of βAS (G. glabra) and ERG1 (Candida albicans) | β-Amyrin (6, 24.50 mg/L) | ||
| Insertion of βAS (G. glabra), ERG1 (C. albicans), IDI (Escherichia coli) Overexpression of ERG9 and ERG20 |
β-Amyrin (6, 36.50 mg/L, 75.50 mg/L *) | ||
| Insertion of βAS (G. glabra), ERG1 (C. albicans), IDI (Escherichia coli) Overexpression of ERG9 and ERG20 Reconstruction of SRE into promoter |
β-Amyrin (6, 85.78 mg/L, 108.60 mg/L *) | ||
| S. cerevisiae W303-1a | Overexpression of tHMG1 and ERG20 | Squalene (4, 165.28 mg/L) | [64] |
| S. cerevisiae WTE | Insertion of mix-AS (Catharanthus roseus) Overexpression of tHMG1 and ERG20 |
β-Amyrin (6, 1.64 mg/L), α-amyrin (7, 5.64 mg/L) | |
| Insertion of mix-AS (Catharanthus roseus) and ERG1 (C. albicans) Overexpression of tHMG1 |
β-Amyrin (6, 24.95 mg/L, 44.92 mg/L *), α-amyrin (7, 97.31 mg/L, 175.15 mg/L *) | ||
| S. cerevisiae ScLCZ08 | Insertion of CrOAS (C. roseus) and AtCPR1 (A. thaliana) | OA (1, 29.49 mg/L), UA (3, 24.58 mg/L) | |
| Insertion of MtOAS (M. truncatula) and LjCPR1 (L. japonicus) | OA (1, 24.34 mg/L), UA (3, 23.37 mg/L) | ||
| Insertion of CrOAS (C. roseus) and LjCPR1 (L. japonicus) | OA (1, 23.91 mg/L), UA (3, 22.96 mg/L) | ||
| Insertion of MtOAS (M. truncatula) and AtCPR1 (A. thaliana) | OA (1, 31.41 mg/L, 155.58 mg/L *), UA (3, 25.85 mg/L, 123.37 mg/L *) | ||
| S. cerevisiae JDY52 | Insertion of GgβAS (G. glabra), MtCYP716A12 (M. truncatula), and AtCPR1 (A. thaliana) | OA (1, 2.5 ± 0.2 mg/L) | [65] |
| Insertion of GgβAS (G. glabra), MtCYP716A12 (M. truncatula), and LjCPR (L. japonicus) | OA (1, 6.3 ± 0.3 mg/L) | ||
| Insertion of GgβAS (G. glabra), MtCYP716A12 (M. truncatula), and GuCPR (G. uralensis) | OA (1, 7.1 ± 0.5 mg/L) | ||
| Insertion of GgβAS (G. glabra), MtCYP716A12, and MtCPR (M. truncatula) | OA (1, 9.0 ± 0.7 mg/L) | ||
| Insertion of GgβAS (G. glabra), MtCYP716A12, and MtCPR (M. truncatula) Knocking out of GAL1 |
OA (1, 41.3 ± 3.4 mg/L) | ||
| Insertion of GgβAS (G. glabra), MtCYP716A12, and MtCPR (M. truncatula) Knocking out of GAL1 and GAL80 |
OA (1, 70.3 ± 7.0 mg/L) | ||
| Insertion of GgβAS (G. glabra), MtCYP716A12, and MtCPR (M. truncatula) Knocking out of GAL1 and GAL80 Overexpression of tHMG1, ERG1, and ERG9 |
OA (1, 186.1 ± 12.4 mg/L, 606.9 ± 9.1 mg/L *) | ||
| S. cerevisiae INVSc1 | Insertion of CYP88D6, CYP72A154 (G. uralensis), and ATR1 (A. thaliana) | GA (2, 20.4 ± 7.7 µg/L), 11-oxo-β-amyrin (15, 0.5 ± 0.1 mg/L), β-amyrin (6, 21.23 ± 1.64 mg/L), 11α-hydroxy-β-amyrin (16), 30-hydroxy-11-oxo-β-amyrin (17), and glycyrrhetaldehyde (18) (trace amount) | [62] |
| S. cerevisiae INVSc1 (diploid) | Insertion of CYP88D6, CYP72A154 (G. uralensis), and ATR1 (A. thaliana) | GA (2, 31.8 ± 6.6 µg/L), 11-oxo-β-amyrin (15, 0.5 ± 0.06 mg/L) | |
| S. cerevisiae SGib | Insertion of CYP88D6, CYP72A154 (G. uralensis), and ATR1 (A. thaliana) | GA (2, 33.7 ± 7.4 µg/L), 11-oxo-β-amyrin (15, 7.5 ± 0.5 mg/L) | |
| Insertion of CYP88D6 and ATR1 (A. thaliana) | 11-oxo-β-amyrin (15, 3.1 ± 0.05 mg/L) | ||
| Insertion of Unigene25647 and ATR1 (A. thaliana) | 11-oxo-β-amyrin (15, 6.0 ± 0.1 mg/L) | ||
| Insertion of CYP72A63, Unigene25647 (G. uralensis), and ATR1 (A. thaliana) | GA (2, 40.9 ± 6.4 µg/L), 11-oxo-β-amyrin (15, 9.16 ± 1.20 mg/L) | ||
| Insertion of CYP72A154, Unigene25647 (G. uralensis), and ATR1 (A. thaliana) | GA (2, 42.3 ± 5.8 µg/L), 11-oxo-β-amyrin (15, 7.22 ± 1.58 mg/L) | ||
| Insertion of CYP72A154, Unigene25647, and GuCPR1 (G. uralensis) | GA (2, 517.4 ± 35.5 µg/L), 11-oxo-β-amyrin (15, 15.3 ± 1.6 mg/L) | ||
| Insertion of CYP72A63, Unigene25647, and GuCPR1 (G. uralensis) | GA (2, 7.4 ± 1.0 mg/L, 18.9 ± 2.0 mg/L *), 11-oxo-β-amyrin (15, 22.6 ± 0.9 mg/L, ~80 mg/L *) | ||
| S. cerevisiae BY-OA | Insertion of CYP716C49 (Crataegus pinnatifida) | Maslinic acid (19, 0.06 mg/L/OD600) | [70] |
| Insertion of CaCYP716C49 (Centella asiatica) | Maslinic acid (19, 0.2 mg/L/OD600) | ||
| Insertion of codon-optimized CaCYP716C49 (C. asiatica) | Maslinic acid (19, 0.45 mg/L/OD600, 384.3 mg/L *) | ||
| Insertion of mix-AS (Eriobotrya japonica), VvCYP716A15, and CPR (Vitis vinifera) | UA (3, 1.76 mg/L/OD600), OA (2, 0.61 mg/L/OD600) | ||
| Insertion of mix-AS (E. japonica), codon-optimized CaCYP716C49 (C. asiatica), VvCYP716A15, and CPR (V. vinifera) | Corosolic acid (20, 0.39 mg/L/OD600, 141.0 mg/L*) | ||
| S. cerevisiae BY4742 | Insertion of AaβAS (Artemisia annua) | β-Amyrin (6, 10.8 ± 1.0 mg/L) | [68] |
| Insertion of CqβAS (Chenopodium quinoa) | β-Amyrin (6, 10.8 ± 1.0 mg/L) | ||
| Insertion of PtβAS (Polygala tenuifolia) | β-Amyrin (6, 9.0 ± 0.7 mg/L) | ||
| Insertion of LjβAS (L. japonicus) | β-Amyrin (6, 8.2 ± 1.0 mg/L) | ||
| Insertion of EtβAS (Euphorbium tirucalli) | β-Amyrin (6, 8.0 ± 0.2 mg/L) | ||
| Insertion of SlβAS (Solanum lycopersicum) | β-Amyrin (6, 2.9 ± 0.3 mg/L) | ||
| Insertion of CYP716AL1 (C. roseus) and ATR2 (A. thaliana) | OA (1, 14.3 ± 1.6 mg/L), erythrodiol (8), and oleanolic aldehyde (9) (trace amount) | ||
| Insertion of CYP716A52v2 (P. ginseng) and ATR2 (A. thaliana) | OA (1, 3.0 ± 0.0 mg/L), erythrodiol (8), and oleanolic aldehyde (9) (trace amount) | ||
| Insertion of SlβAS (S. lycopersicum), CYP716AL1 (C. roseus), and ATR2 (A. thaliana) | OA (1, 3.9 ± 0.2 mg/L) | ||
| Insertion of SlβAS (S. lycopersicum), CYP716A52v2 (P. ginseng), and ATR2 (A. thaliana) | OA (1, 2.8 ± 0.0 mg/L) | ||
| Insertion of AaβAS (A. annua), CYP716AL1 (C. roseus), and ATR2 (A. thaliana) | OA (1, 8.5 ± 0.2 mg/L) | ||
| Yarrowia lipolytica ATCC 201249 | Insertion of βAS (C. roseus) | β-Amyrin (6, 16.0 mg/L) | [69] |
| Insertion of βAS (C. roseus), CYP716A12 (M. truncatula), and ATR1 (A. thaliana) | OA (1, 16.3 mg/L) | ||
| Insertion of βAS (C. roseus), CYP716A12 (M. truncatula), and ATR1(A. thaliana) Overexpression of ERG1, ERG9, ERG20, tHMG1 |
OA (1, 92.1 mg/L) | ||
| Insertion of βAS (C. roseus) and CYP716A2-linker (GSTSSG)-t46ATR1 Overexpression of ERG9, ERG20, and tHMG1 |
OA (1, 129.9 mg/L, 540.7 mg/L*) |
Note: * Yield of the compound in a 5-L fermenter. ** The number in bold is the number of the compound structural formula. GA, glycyrrhetinic acid; OA, oleanolic acid; UA, ursolic acid.