CRISPR-assisted rational flux-tuning and arrayed CRISPRi screening of an l-proline exporter for l-proline hyperproduction

Jiao Liu, Moshi Liu, Tuo Shi, Guannan Sun, Ning Gao, Xiaojia Zhao, Xuan Guo, Xiaomeng Ni, Qianqian Yuan, Jinhui Feng, Zhemin Liu, Yanmei Guo, Jiuzhou Chen, Yu Wang (), Ping Zheng () and Jibin Sun
Additional contact information
Jiao Liu: Chinese Academy of Sciences
Moshi Liu: Chinese Academy of Sciences
Tuo Shi: Chinese Academy of Sciences
Guannan Sun: Chinese Academy of Sciences
Ning Gao: Chinese Academy of Sciences
Xiaojia Zhao: Chinese Academy of Sciences
Xuan Guo: Chinese Academy of Sciences
Xiaomeng Ni: Chinese Academy of Sciences
Qianqian Yuan: Chinese Academy of Sciences
Jinhui Feng: Chinese Academy of Sciences
Zhemin Liu: Chinese Academy of Sciences
Yanmei Guo: Chinese Academy of Sciences
Jiuzhou Chen: Chinese Academy of Sciences
Yu Wang: Chinese Academy of Sciences
Ping Zheng: Chinese Academy of Sciences
Jibin Sun: Chinese Academy of Sciences

Nature Communications, 2022, vol. 13, issue 1, 1-16

Abstract: Abstract Development of hyperproducing strains is important for biomanufacturing of biochemicals and biofuels but requires extensive efforts to engineer cellular metabolism and discover functional components. Herein, we optimize and use the CRISPR-assisted editing and CRISPRi screening methods to convert a wild-type Corynebacterium glutamicum to a hyperproducer of l-proline, an amino acid with medicine, feed, and food applications. To facilitate l-proline production, feedback-deregulated variants of key biosynthetic enzyme γ-glutamyl kinase are screened using CRISPR-assisted single-stranded DNA recombineering. To increase the carbon flux towards l-proline biosynthesis, flux-control genes predicted by in silico analysis are fine-tuned using tailored promoter libraries. Finally, an arrayed CRISPRi library targeting all 397 transporters is constructed to discover an l-proline exporter Cgl2622. The final plasmid-, antibiotic-, and inducer-free strain produces l-proline at the level of 142.4 g/L, 2.90 g/L/h, and 0.31 g/g. The CRISPR-assisted strain development strategy can be used for engineering industrial-strength strains for efficient biomanufacturing.

Date: 2022
References: View references in EconPapers View complete reference list from CitEc
Citations: View citations in EconPapers (1)

Downloads: (external link)
https://www.nature.com/articles/s41467-022-28501-7 Abstract (text/html)

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:13:y:2022:i:1:d:10.1038_s41467-022-28501-7

Ordering information: This journal article can be ordered from
https://www.nature.com/ncomms/

DOI: 10.1038/s41467-022-28501-7

Access Statistics for this article

Nature Communications is currently edited by Nathalie Le Bot, Enda Bergin and Fiona Gillespie

More articles in Nature Communications from Nature
Bibliographic data for series maintained by Sonal Shukla () and Springer Nature Abstracting and Indexing ().