Engineering CrtW and CrtZ for improving biosynthesis of astaxanthin in Escherichia coli

Li, Di; Li, Yang; Xu, Jiaoyang; Li, Qingyan; Tang, Ji‐Xin; Jia, Shiru; Bi, Chen; Zhu, Xinna; Zhang, Xueli

doi:10.1016/s1875-5364(20)60005-x

Cited by 16 publications

(21 citation statements)

References 27 publications

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“…agglomerans using an RBS library, followed by the use of multicopy expression of crtW* and crtZ . The best chassis produced astaxanthin as the predominant carotenoid (99%) with a specific production of 0.88 g/L in fed-batch fermentation . The crtW* mutant with A6T, T105A, and L239M mutations has improved 5.35-fold astaxanthin production compared with the wild-type control .…”

Section: Discussionmentioning

confidence: 99%

“…The best chassis produced astaxanthin as the predominant carotenoid (99%) with a specific production of 0.88 g/L in fed-batch fermentation . The crtW* mutant with A6T, T105A, and L239M mutations has improved 5.35-fold astaxanthin production compared with the wild-type control . Therefore, crtW* and crtZ were used to construct a pathway for astaxanthin biosynthesis in this study.…”

Section: Discussionmentioning

confidence: 99%

“…1.2244 (GenBank accession number HQ003247) using the primer pair CrtZ-RBS-99A-CPEC-F/CrtZ-taa-R (Table S1), and crtW* was codon-optimized according to the sequence of crtW from Brevundimonas sp. SD212 (GenBank accession number AB181388) and optimized by random mutation and directed evolution . Its coding sequence was amplified using the primer pair CrtW-RBS-crtZ-F/CrtW-CPEC-R, and the plasmid backbone was amplified from pSC103 using the primer pair 99A-CPEC-F/99A-CPEC-R. Then, the three PCR fragments were ligated using the Seamless Cloning Kit (Beyotime), resulting in the plasmid psc104-crtZW* (Figure S1B).…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Coordinated Expression of Astaxanthin Biosynthesis Genes for Improved Astaxanthin Production in Escherichia coli

Gong

Wang

Tang

et al. 2020

J. Agric. Food Chem.

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Astaxanthin has great potential commercial value in the feed, cosmetics, and nutraceutical industries due to its strong antioxidant capacity. In this study, the Escherichia coli strain CAR026 with completely balanced metabolic flow was selected as the starting strain for the production of astaxanthin. The expression of β-carotene ketolase (CrtW) and β-carotene hydroxylase (CrtZ), which catalyze the conversion of β-carotene to astaxanthin, was coordinated, and a bottleneck was eliminated by increasing the copy number of crtY in CAR026. The resulting strain Ast007 produced 21.36 mg/L and 4.6 mg/g DCW of astaxanthin in shake flasks. In addition, the molecular chaperone genes groES-groEL were regulated to further improve the astaxanthin yield. The best strain Gro-46 produced 26 mg/L astaxanthin with a yield of 6.17 mg/g DCW in shake flasks and 1.18 g/L astaxanthin after 60 h of fermentation under fed-batch conditions. To the best of our knowledge, this is the highest astaxanthin obtained using engineered E. coli to date.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Coordinated Expression of Astaxanthin Biosynthesis Genes for Improved Astaxanthin Production in Escherichia coli

Gong

Wang

Tang

et al. 2020

J. Agric. Food Chem.

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“…For instance, replacement of the CrtZ gene promoter with a stronger one increased the enzyme level ratio of CrtZ to CrtW and yielded 30.4% increase in astaxanthin [ 136 ]. In order to finely optimize the ratio the two enzymes, screening of several RBS libraries obtained through introducing six random nucleotides to the RBS region of these genes resulted in optimized cassette with a titer of 3.46 mg/g DCW and 99% astaxanthin [ 137 ]. Further increase by 67.6% was obtained by multiple integration of the optimized cassette.…”

Section: Strategies For Microbial Cell Factories Optimizationmentioning

confidence: 99%

“…Another triple mutant (A6T, T105A and L239 M) of CrtW from Brevundimonas sp. SD212 enhanced astaxanthin level by 5.35 fold in E. coli [ 137 ]. The mutant showed enhanced conversion of the intermediates and astaxanthin reached 92% of the total carotenoids produced by this strain.…”

Section: Strategies For Microbial Cell Factories Optimizationmentioning

confidence: 99%

Optimization of microbial cell factories for astaxanthin production: Biosynthesis and regulations, engineering strategies and fermentation optimization strategies

Basiony

Ouyang

Wang

et al. 2022

Synthetic and Systems Biotechnology

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Rapid construction of Escherichia coli chassis with genome multi‐position integration of isopentenol utilization pathway for efficient and stable terpenoid accumulation

Wang

Huang

et al. 2023

Biotechnology Journal

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The isopentenol utilization pathway (IUP) is potential in terpenoids synthesis. This study aimed to construct IUP‐employed Escherichia coli chassis for stably synthesizing terpenoids. As to effectiveness, promotor engineering strategy was employed to regulate IUP expression level, while ribosome‐binding site (RBS) library of the key enzyme was constructed for screening the optimal RBS, followed by optimization of concentration of inducer and substrates, the titer of reporting production, lycopene, from 0.087 to 8.67 mg OD600−1. As about stability, the IUP expression cassette was integrated into the genome through transposition tool based on CRISPR‐associated transposases. Results showed that the strain with 13 copies produced 1.78‐fold lycopene titer that of the controlled strain with IUP‐harbored plasmid, and it exhibited stable expression after ten successions while the plasmid loss was observed in the controlled strain in the 3rd succession. This strategy provides valuable information for rapid construction of highly effective and stable chassis employing IUP for terpenoids production.

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Engineering CrtW and CrtZ for improving biosynthesis of astaxanthin in Escherichia coli

Cited by 16 publications

References 27 publications

Coordinated Expression of Astaxanthin Biosynthesis Genes for Improved Astaxanthin Production in Escherichia coli

Coordinated Expression of Astaxanthin Biosynthesis Genes for Improved Astaxanthin Production in Escherichia coli

Optimization of microbial cell factories for astaxanthin production: Biosynthesis and regulations, engineering strategies and fermentation optimization strategies

Rapid construction of Escherichia coli chassis with genome multi‐position integration of isopentenol utilization pathway for efficient and stable terpenoid accumulation

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