Heterologous expression and metabolic engineering tools for improving terpenoids production

Ma, Chuanteng; Zhang, Kaijin; Zhang, Xianyan; Liu, Guowei; Zhu, Tianjiao; Che, Qian; Li, Dehai; Zhang, Guojian

doi:10.1016/j.copbio.2021.02.008

Cited by 22 publications

(19 citation statements)

References 63 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…As acetyl-CoA is the direct precursor of the mevalonate pathway, the overexpression of the acetyl-CoA synthase gene ( ACS* , a feedback inhibition insensitive mutant from Salmonella enterica) should benefit the synthesis of α-santalene. The integration of ACS* into the genome of STE-6 resulted in the construction of STE-7, whose production of α-santalene was further increased to 282.4 ± 44.1 mg/L.…”

Section: Resultsmentioning

confidence: 98%

“…Simultaneous overexpression of tHMG1 and IDI1-ERG20 (STE-6) further increased the production of α-santalene to 246.9 ± 33.8 mg/L, which was about 2.74-fold higher than that of STE-1 (Figure 3). As acetyl-CoA is the direct precursor of the mevalonate pathway, 37 the overexpression of the acetyl-CoA synthase gene (ACS*, a feedback inhibition insensitive mutant from Salmonella enterica) 38 should benefit the synthesis of α-santalene. The integration of ACS* into the genome of STE-6 resulted in the construction of STE-7, whose production of α-santalene was further increased to 282.4 ± 44.1 mg/L.…”

Section: ■ Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Establishing Komagataella phaffii as a Cell Factory for Efficient Production of Sesquiterpenoid α-Santalene

Zuo

Xiao

Gao

et al. 2022

J. Agric. Food Chem.

View full text Add to dashboard Cite

Santalene, a major component of the sandalwood essential oil, is a typical representative of sesquiterpenes and has important applications in medicine, food, flavors, and other fields. Due to the limited supply of natural sandalwood resources, there is a growing interest in engineering microbial cell factories for the mass production of santalene. In the present study, Komagataella phaffii (also known as Pichia pastoris) was established as a cell factory for high-level production of α-santalene for the first time. The metabolic fluxes were rewired toward α-santalene biosynthesis through the optimization of promoters to drive the expression of the α-santalene synthase (SAS) gene, overexpression of the key mevalonate pathway genes (i.e., tHMG1, IDI1, and ERG20), and multicopy integration of the SAS expression cassette. In combination with medium optimization and bioprocess engineering, the optimal strain (STE-9) was able to produce α-santalene with a titer as high as 829.8 ± 70.6 mg/L, 4.4 ± 0.3 g/L, and 21.5 ± 1.6 g/L in a shake flask, batch fermenter, and fed-batch fermenter, respectively. These represented the highest production of α-santalene ever reported, highlighting the advantages of K. phaffii cell factories for the production of terpenoids and other natural products.

show abstract

Section: Resultsmentioning

confidence: 98%

Section: ■ Resultsmentioning

confidence: 99%

Establishing Komagataella phaffii as a Cell Factory for Efficient Production of Sesquiterpenoid α-Santalene

Zuo

Xiao

Gao

et al. 2022

J. Agric. Food Chem.

View full text Add to dashboard Cite

show abstract

“…In turn, IPP and DMAPP can be generated through the methylerythritol 4-phosphate (MEP) pathway or the mevalonate (MVA) pathway Saccharomyces cerevisiae utilizes a native MVA pathway to synthesize IPP and DMAPP, which is tightly regulated by the host . Some metabolic engineering approaches including downregulation of the squalene synthase enzyme (EGR9), knockout of the transcription factor (ROX1), and overexpression of the truncated version of 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase (tHMG1) were subjected to the (+)-nootkatone production by S.…”

Section: Introductionmentioning

confidence: 99%

“…10 Saccharomyces cerevisiae utilizes a native MVA pathway to synthesize IPP and DMAPP, which is tightly regulated by the host. 11 Some metabolic engineering approaches including downregulation of the squalene synthase enzyme (EGR9), knockout of the transcription factor (ROX1), and overexpression of the truncated version of 3-hydroxy-3methylglutaryl-CoA (HMG-CoA) reductase (tHMG1) were subjected to the (+)-nootkatone production by S. cerevisiae. 9,12 However, the final (+)-nootkatone titer was too low to be detected by gas chromatography (GC).…”

Section: Introductionmentioning

confidence: 99%

Probing the Synergistic Ratio of P450/CPR To Improve (+)-Nootkatone Production in Saccharomyces cerevisiae

Cha

Wen

et al. 2022

J. Agric. Food Chem.

View full text Add to dashboard Cite

(+)-Nootkatone is an expensive sesquiterpene substance found in grapefruit peels and the heartwood of yellow cedar. It can be used as a food additive, perfume, and insect repellent; therefore, its highly efficient production is greatly needed. However, the low catalytic efficiency of the membrane-anchored cytochrome P450/P450 reductase system (HPO/AtCPR) is the main challenge and limits the production of (+)-nootkatone. We developed an effective high-throughput screening system based on cell wall destruction to probe the optimal ratio of HPO/AtCPR, which achieved a twofold elevation in (+)-valencene oxidation in Saccharomyces cerevisiae. An engineered strain PK2RI-AtC/Hm6A was constructed to realize de novo (+)-nootkatone production by a series of metabolic engineering strategies. In biphasic fed-batch fermentation, maximum titers of 3.73 and 1.02 g/L for (+)-valencene and (+)-nootkatone, respectively, were achieved. The dramatically improved performance of the constructed S. cerevisiae provides an excellent approach for economical production of (+)-nootkatone from glucose.

show abstract

“…In addition, E. coli possesses several attractive traits, such as fast growth and its ease of genetic manipulation (Zhang et al 2019 ). Another advantage is the minimal secondary metabolite background of E. coli cells, which facilitates the recovery of heterologously produced compounds (Ma et al 2021 ).…”

Section: Introductionmentioning

confidence: 99%

Biocatalytic production of the antibiotic aurachin D in Escherichia coli

2022

View full text Add to dashboard Cite

Aurachin D is a potent inhibitor of cytochrome bd oxidases, which are potential targets in the treatment of infectious diseases. In this study, our aim was to improve the biocatalytic production of aurachin D from a quinolone precursor molecule with recombinant Escherichia coli cells expressing the biosynthesis enzyme AuaA. In order to achieve a high-level production of this membrane-bound farnesyltransferase in E. coli, the expression of the auaA gene was translationally coupled to an upstream cistron in accordance with a bicistronic design (BCD) strategy. Screening of various BCD elements led to the identification of optimized auaA expression cassettes, which increased the aurachin D titer in E. coli up to 29-fold in comparison to T7-mediated expression. This titer could be further raised by codon optimization of auaA and by introducing the mevalonate pathway into the production strain. The latter measure was intended to improve the availability of farnesyl pyrophosphate, which is needed as a cosubstrate for the AuaA-catalyzed reaction. In sum, the described efforts resulted in a strain producing aurachin D with a titer that is 424 times higher than that obtained with the original, non-optimized expression host. Graphical Abstract

show abstract

Heterologous expression and metabolic engineering tools for improving terpenoids production

Cited by 22 publications

References 63 publications

Establishing Komagataella phaffii as a Cell Factory for Efficient Production of Sesquiterpenoid α-Santalene

Establishing Komagataella phaffii as a Cell Factory for Efficient Production of Sesquiterpenoid α-Santalene

Probing the Synergistic Ratio of P450/CPR To Improve (+)-Nootkatone Production in Saccharomyces cerevisiae

Biocatalytic production of the antibiotic aurachin D in Escherichia coli

Contact Info

Product

Resources

About