High-Yielding Terpene-Based Biofuel Production in <i>Rhodobacter capsulatus</i>

Zhang, Yang; Song, Xiaohui; Lai, Yumeng; Mo, Qiwen

doi:10.1021/acssynbio.1c00146

Cited by 22 publications

(9 citation statements)

References 45 publications

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“…Deletion of zwf increased the β‐elemene production by 11.12% relative to strain EL‐7 by increased flux toward the TCA cycle. Our result was consistent with recent studies that saw a 91% improvement in bisabolene production after zwf deletion (Zhang et al, 2021) and an enhancement in lycopene production after zwf inactivation (Zhou et al, 2013). The PPP is a crucial energy metabolism pathway, and the overexpression of zwf has been associated with increased NADPH supply for α‐farnesene synthesis (Chen et al, 2023).…”

Section: Discussionsupporting

confidence: 94%

Engineering Escherichia coli BL21 (DE3) for high‐yield production of germacrene A, a precursor of β‐elemene via combinatorial metabolic engineering strategies

Fordjour

Liu

Hao

et al. 2023

Biotech & Bioengineering

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Abstractβ‐elemene is one of the most commonly used antineoplastic drugs in cancer treatment. As a plant‐derived natural chemical, biologically engineering microorganisms to produce germacrene A to be converted to β‐elemene harbors great expectations since chemical synthesis and plant isolation methods come with their production deficiencies. In this study, we report the design of an Escherichia coli cell factory for the de novo production of germacrene A to be converted to β‐elemene from a simple carbon source. A series of systematic approaches of engineering the isoprenoid and central carbon pathways, translational and protein engineering of the sesquiterpene synthase, and exporter engineering yielded high‐efficient β‐elemene production. Specifically, deleting competing pathways in the central carbon pathway ensured the availability of acetyl‐coA, pyruvate, and glyceraldehyde‐3‐phosphate for the isoprenoid pathways. Adopting lycopene color as a high throughput screening method, an optimized NSY305N was obtained via error‐prone polymerase chain reaction mutagenesis. Further overexpression of key pathway enzymes, exporter genes, and translational engineering produced 1161.09 mg/L of β‐elemene in a shake flask. Finally, we detected the highest reported titer of 3.52 g/L of β‐elemene and 2.13 g/L germacrene A produced by an E. coli cell factory in a 4‐L fed‐batch fermentation. The systematic engineering reported here generally applies to microbial production of a broader range of chemicals. This illustrates that rewiring E. coli central metabolism is viable for producing acetyl‐coA‐derived and pyruvate‐derived molecules cost‐effectively.

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Section: Discussionsupporting

confidence: 94%

Engineering Escherichia coli BL21 (DE3) for high‐yield production of germacrene A, a precursor of β‐elemene via combinatorial metabolic engineering strategies

Fordjour

Liu

Hao

et al. 2023

Biotech & Bioengineering

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“…lipolytica cell factory reported to date. Although the highest reported bisabolene titer of 9.8 g/L was achieved in Rhodobacter capsulatus through fed-batch bioreactor fermentation, it is worth noting that our study provides a consolidated process toward bisabolene synthesis from cost-free food wastes. In practice, the production capability of the microbial cell factory is highly dependent on fermentation parameters such as temperature, pH, dissolved oxygen, and initial cell density.…”

Section: Resultsmentioning

confidence: 88%

Mitochondrial Engineering of Yarrowia lipolytica for Sustainable Production of α-Bisabolene from Waste Cooking Oil

Zhu

Zhao

Zhang

et al. 2022

ACS Sustainable Chem. Eng.

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Metabolic engineering of yeasts for terpenoid production has mostly focused on the cytoplasm, whereas harnessing their organelles as subcellular factories has been overlooked. Herein, the farnesyl diphosphate synthetic pathway and α-bisabolene synthase were compartmentalized into the oleaginous yeast Yarrowia lipolytica's mitochondria to enable high-level α-bisabolene production. Through comprehensive metabolic engineering approaches, we exploited the potential and capability of the mitochondria as a subcellular factory to achieve 257.4 mg/L of α-bisabolene production from glucose. By combining mitochondrial and cytoplasmic engineering, we further boosted the α-bisabolene titer to 765.1 mg/L by utilizing waste cooking oil as the sole carbon source. Finally, the α-bisabolene titer of the resulting strain reached 1058.1 mg/L in a 5 L bioreactor, which is the highest titer in the engineered Y. lipolytica cell factory reported to date. Overall, our study has provided valuable insights into the mitochondrial engineering of Y. lipolytica for sustainable and green production of valuable compounds.

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“…lipolytica , are the favored hosts for studies on the production of valuable terpenoids using metabolic engineering and synthetic biology. In recent years, some emerging microbial hosts have shown robust potential for the production of precious terpenoids due to their unique advantages. ,− It is well known that the antibacterial activity and toxicity of most monoterpenoids hinder the wide application of microorganisms in their production. Rather than using various engineering strategies to improve the tolerance and carbon flux of the chassis, other microorganisms should be selected because natural genetic advantages can provide a more suitable metabolic framework for the biosynthesis of target products.…”

Section: Results and Discussionmentioning

confidence: 99%

Tuning Geraniol Biosynthesis via a Novel Decane-Responsive Promoter in Candida glycerinogenes

Zhao

Wang

et al. 2022

ACS Synth. Biol.

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Geraniol is a rose-scented monoterpene with significant commercial and industrial value in medicine, condiments, cosmetics, and bioenergy. Here, we first targeted geraniol as a reporter metabolite and explored the suitability and potential of Candida glycerinogenes as a heterologous host for monoterpenoid production. Subsequently, dual-pathway engineering was employed to improve the production of geraniol with a geraniol titer of 858.4 mg/L. We then applied a synthetic hybrid promoter approach to develop a decane-responsive hybrid promoter based on the native promoter P GAP derived from C. glycerinogenes itself. The hybrid promoter was able to be induced by n-decane with 3.6 times higher transcriptional intensity than the natural promoter P GAP . In particular, the hybrid promoter effectively reduces the conflict between cell growth and product formation in the production of geraniol. Ultimately, 1194.6 mg/L geraniol was obtained at the shake flask level. The strong and tunable decane-responsive hybrid promoter developed in this study provides an important tool for fine regulation of toxic terpenoid production in cells.

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High-Yielding Terpene-Based Biofuel Production in Rhodobacter capsulatus

Cited by 22 publications

References 45 publications

Engineering Escherichia coli BL21 (DE3) for high‐yield production of germacrene A, a precursor of β‐elemene via combinatorial metabolic engineering strategies

Engineering Escherichia coli BL21 (DE3) for high‐yield production of germacrene A, a precursor of β‐elemene via combinatorial metabolic engineering strategies

Mitochondrial Engineering of Yarrowia lipolytica for Sustainable Production of α-Bisabolene from Waste Cooking Oil

Tuning Geraniol Biosynthesis via a Novel Decane-Responsive Promoter in Candida glycerinogenes

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