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Table 1 Some representative examples of limonene production in engineered microbial hosts

From: Engineering the oleaginous yeast Yarrowia lipolytica to produce limonene from waste cooking oil

Host Product Titer Yield Productivity Strategy References
E. coli l-Limonene 430 mg/L 5.97 mg/L/h A plasmid with a truncated GPP synthase and limonene synthase gene was co-expressed with a plasmid increasing the MK and PMK levels (MBI-f operon) and replacing the HMGS and HMGR from S. cerevisiae (MevT operon) with those from S. aureus (MTSA operon) using different IPTG levels [28]
l-Limonene 2700 mg/Lorg 60 mg/Lorg/h (1) Truncated LS gene and truncated GPPS2 gene with all the genes of the MVA pathway, that convert endogenous acetyl-CoA into IPP and DMAPP from S. cerevisiae was co-expressed. (2) A two-liquid phase fed-batch fermentation with glycerol as carbon source were carried out in 3.1 L stirred-tank reactors [35]
l-Limonene 75.12 mg/L 3.13 mg/L/h (1) Juice from sugar beets was utilized as a novel feedstock. (2) A synthetic DXP operon (dxs-ipphp-gpps) encoding three rate-limiting enzymes from the MEP pathway was co-expressed with limonene synthases gene [75]
S. cerevisiae d-Limonene and l-limonene 0.12 mg/L d-limonene and 0.49 mg/L l-limonene 1.67 µg/L/h d-limonene and 6.8 µg/L/h l–limonene The truncated limonene synthases with Cytochrome P450 reductase (CPR1) (Gal1 promoter) were co-expressed, and a setup was built which allowed trapping of exhaust volatiles during growth [33]
Y. lipolytica d-Limonene and l-limonene 11.705 mg/L d-limonene and 11.088 mg/L l-limonene 0.443 mg/g DCW for d-limonene and 0.385 mg/g DCW for l-limonene 0.0542 mg/L/h d-limonene and 0.0513 mg/L/h l-limonene (1) The heterologous d-limonene synthase from Citrus limon and l-limonene synthase from Mentha spicata with ten homologous genes involved in the MVA pathway were overexpressed, respectively. (2) Fermentation conditions of engineered strains were optimized. (3) Fed-batch fermentation of engineered strains was used. (4) WCO was used as carbon resource This work
d-Limonene 23.56 mg/L 1.36 mg/g DCW 0.33 mg/L/h The truncated genes tLS and tNDPS1 with HMG1 and ERG12 were co-expressed [72]
Anabaena sp. PCC 7120 l-Limonene 172.7 µg/L/48 h A synthetic DXP operon (dxs-ipphp-gpps) encoding three rate-limiting enzymes from the MEP pathway was co-expressed with limonene synthases gene [76]
Synechococcus sp. PCC 7002 l-Limonene 4 mg/L 0.042 mg/L/h The (−)-4S-limonene synthase gene was expressed [30]
Synechocystis sp. PCC 6803 d-Limonene 0.564 mg/L 2.33 µg/L/h Introducing an additional copy of three genes (dxs, crtE and ipi) under the control of a strong trc promoter [77]