Table 1 Typical protein engineering strategies for terpenoid biosynthesis
From: Toward improved terpenoids biosynthesis: strategies to enhance the capabilities of cell factories
Strain | Product | Description | Outcome | Refs. |
|---|---|---|---|---|
Protein engineering | ||||
 E. coli | Viridiflorol | Random mutation by error-prone PCR and enzyme truncation on viridiflorol synthase | 25.7 g/L | Shukal et al. (2019) |
 E. coli | Levopimaradiene | Combinatorial evolution in geranylgeranyl diphosphate synthase and levopimaradiene synthase | 700 mg/L | Leonard et al. (2010) |
 E. coli | Taxa-4(20)-11(12)-diene | Site-saturated mutagenesis on taxadiene synthase | 2.4-fold | Edgar et al. (2017) |
 E. coli | Amorpha-4,11-diene | Mutability landscape engineering of amorpha-4,11-diene synthase | Fourfold | Abdallah et al. (2018) |
 E. coli | Acetyl-CoA | Site-directed mutagenesis of citrate synthase (gltA) | 0.24 g/g glucose | Tovilla-Coutiño et al. (2020) |
 E. coli | 8-epi-cedrol | Fusion of farnesyl pyrophosphate and Epi-cedrol synthase | 1.084 mg/L | Navale et al. (2019) |
 E. coli | Astaxanthin | Fusion of crtZ and crtW | 1.4-fold increment | Nogueira et al. (2019) |
 E. coli | Steviol glycosides | Fusion of CYP714A2 and P450 reductase 2 (AtCPR2) from A. thaliana | 38.4 ± 1.7 mg/L | Moon et al. (2020) |
 E. coli | Lycopene | Enzyme engineering of mevalonate 3-kinase via single amino acid mutations into 5-phosphomevalonate 3-kinase | N. A | Motoyama et al. (2019) |
 E. coli | Albaflavenol and (1R)-6β-hydroxycineole | Fusion of terpene synthases and cytochrome P450 | 90- and 5.4-fold improvement, respectively | Wang et al. (2021d) |
 E. coli | Geraniol | Fusion tagging of geraniol synthase with a laboratory evolved N-terminal of a chloramphenicol acetyltransferase leader sequence | 2124.1 mg/L | Wang et al. (2021c) |
 E. coli | (−)-Carvone | Optimization of pathway enzymes with quantification concatemer (QconCat) method | 15-fold | Yoshida et al. (2021) |
 E. coli | Pinene | Computer-aided directed evolution of geranyl diphosphate and pinene synthase | N. A | Chen et al. (2021) |
 E. coli | Nerol and borneol | Site-directed mutagenesis of truncated bornyl diphosphate synthase from Lippia dulcis | 966.55 mg/L and 87.20 mg/L, respectively | Lei et al. (2021) |
 E. coli | (1R)-6β-Hydroxycineole and albaflavenone | Fusion of 1,8-cineole synthase and cytochrome P450 enzyme CYP176A1 for as well as fusion of epi-isozizaene synthase with CYP170A1 | 5.4-, 2.3-fold, respectively | Wang et al. (2021d) |
 E. coli | Isopentenol | Improved activity of phosphomevalonate decarboxylase (PMD) in an isopentenyl diphosphate (IPP)-bypass | 2.4-fold | Kang et al. (2017) |
 S. cerevisiae | Geranylgeraniol | Overexpression of fused geranylgeranyl pyrophosphate synthase from Pantoea agglomerans and farnesyl diphosphate synthase (PaGGPPS-Erg20) and fused geranylgeranyl pyrophosphate synthase from Pantoea agglomerans and diacylglycerol diphosphate phosphate (PaGGPPS-DPP1) | 5.07 g/L | Wang et al. (2021b) |
 S. cerevisiae | Miltiradiene | Engineering a chimeric protein through the fusion of terpene synthase 1 from Coleus forskohlii and kaurene synthases-like from Salvia miltiorrhia | 3.5 g/L | Hu et al. (2020a) |
 S. cerevisiae | Isoprene | Directed evolution of isoprene synthase | 3.7 g/L | Wang et al. (2017) |
 S. cerevisiae | Lycopene | Inactivation of lycopene cyclase activity of crtYB via directed evolution and improved activity of crtE | 1.61 g/L | Xie et al. (2015) |
 S. cerevisiae | Lycopene | Directed evolution on isopentenyl diphosphate isomerase (idi) | 1.2 g/L | Chen et al. (2018a) |
 S. cerevisiae | Lycopene and β-carotene | Screening of efficient crt variants from Sporidiobolus pararoseus TBRC-BCC 63403 | Four and sevenfold, respectively | Watcharawipas et al. (2021) |
 S. cerevisiae | Carotenoid | A bidomain fusion of crtI and crtB and a tridomain fusion of crtI, crtB, and crtY | Twofold | Rabeharindranto et al. (2019) |
 S. cerevisiae | α-farnesene | Screening of an α-farnesene synthase with enhanced catalytic activity | 10.4 g/L | Wang et al. (2021a) |
 S. cerevisiae | d-Limonene | Fusion of truncated limonene synthase from Citrus limon and truncated neryl diphosphate synthase | 23.7 mg/L | Hu et al. (2020b) |
 S. cerevisiae | Diterpenoids | Fusion of Erg20WW, Erg20 and BTS1 | 3.2- and 2.3-fold | Dong et al. (2020a) |
 S. cerevisiae | Geraniol | Fusion of Erg20WW and geraniol synthase | 15% improvement | Jiang et al. (2017) |
 S. cerevisiae | Germacrene A | Fusion of farnesyl pyrophosphate synthase and germacrene A synthase | 190.7 mg/L | Hu et al. (2017) |
 S. cerevisiae | ( +)-Nootkatone | Fusion of (+)-valencene synthase and farnesyl diphosphate synthase | 59.78 mg/L | Meng et al. (2020a) |
S. cerevisiae | Zerumbone | Fusion of cytochrome P450 reductase from A. thaliana and α-humulene 8-hydroxylase (CYP71BA1) | 40 mg/L | Zhang et al. (2018a) |
 S. cerevisiae | Sabinene | N-terminal truncation of sabinene synthase | 96% improvement | Jia et al. (2020) |
 S. cerevisiae | Linalool | N-terminal SKIK tagging of engineered linalool synthase and RIAD/RIDD linking of SKIK linalool synthase and ERG20WW | 42% improvement | Zhou et al. (2021) |
 S. cerevisiae | Lutein | Localization of lycopene ε-cyclase to the cell membrane through fusion technology | N. A | Bian et al. (2021) |
 S. cerevisiae | Violaxanthin | N-terminal truncation of zeaxanthin epoxidases | 7.3 mg/g DCW | Cataldo et al. (2020) |
 S. cerevisiae | α-terpineol | Fusion of truncated α-terpineol synthase with Erg20WW | 21.88 mg/L | Zhang et al. (2019) |
 Y. lipolytica | (+)-nootkatoone | Enzyme fusion of truncated NADPH-cytochrome P450 reductase and (+)-nootkatoone synthase coupled with another optimization | 20.5-fold improvement | Guo et al. (2018) |
 Y. lipolytica | Gibberellin (GA3) | Protein engineering and overexpression of pathway enzymes | 12 mg/L | Kildegaard et al. (2021) |
 Y. lipolytica | α-farnesene | Fusion of Erg20 and farnesyl synthase with lipid as the carbon source | 10.2 g/L | Liu et al. (2021d) |
 Pantoea ananatis | Linalool | Improved soluble expression of (S)-linalool synthase by N-terminal fusion with halophilic β-lactamase hexahistidine | 10.9 g/L | Nitta et al. (2021) |
 C. glutamicum | Astaxanthin | Fusion of membrane-bound crtW and crtZ | 3.1 mg/g DCW | Henke and Wendisch (2019) |
 Bacillus subtilis | Amorphadiene | Site-directed mutagenesis of amorphadiene synthase with a one-plasmid CRISPR–Cas9 editing system | 116 mg/L | Song et al. (2021) |
 Cyanobacteria | Isoprene | Fusion isoprene synthase with the native promoter cpcB | 28.9 µg/L/h | Chaves et al. (2017) |