Journal
ENVIRONMENTAL SCIENCE & TECHNOLOGY
Volume 54, Issue 6, Pages 3599-3608Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.est.9b06378
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Funding
- National Key Research and Development Program of China [2018YFA0901301]
- National Natural Science Foundation of China [21590812, 21806160, 51821006]
- Program for Changjiang Scholars and Innovative Research Team in University of the Ministry of Education of China
- Fundamental Research Funds for the Central Universities [WK 2060000002]
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Pursuing efficient approaches to promote the extracellular electron transfer (EET) of extracellular respiratory bacteria is essential to their application in environmental remediation and waste treatment. Here we report a new strategy of tuning electron flux by clustered regularly interspaced short palindromic repeat (CRISPR)-ddAsCpf1-based rediverting (namely STAR) to enhance the EET capacity of Shewanella oneidensis MR-1, a model extracellular respiratory bacterium widely present in the environment. The developed CRISPR-ddAsCpf1 system enabled approximately 100% gene repression with the green fluorescent protein (GFP) as a reporter. Using a WO3 probe, 10 representative genes encoding for putative competitive electron transfer proteins were screened, among which 7 genes were identified as valid targets for EET enhancement. Repressing the valid genes not only increased the transcription level of the L-lactate metabolism genes but also affected the genes involved in direct and indirect EET. Increased riboflavin production was also observed. The feasibility of this strategy to enhance the bioreduction of methyl orange, an organic pollutant, and chromium, a typical heavy metal, was demonstrated. This work implies a great potential of the STAR strategy with the CIRPSR-ddAsCpf1 system for enhancing bacterial EET to favor more efficient environmental remediation applications.
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