Title:
HIGH-THROUGHPUT METHODS FOR ISOLATING AND CHARACTERIZING AMMONIUM-EXCRETING MUTANT LIBRARIES GENERATED BY CHEMICAL MUTAGENESIS
Document Type and Number:
WIPO Patent Application WO/2020/219893
Kind Code:
A3
Abstract:
The present disclosure provides high-throughput methods for rapidly mutagenizing, screening, and targeting candidate microbes that are capable of fixing atmospheric nitrogen in the presence of exogenous nitrogen. The methods utilize a microbial biosensor capable of detecting the presence/absence of ammonium and/or glutamine in a composition and signaling with a fluorescent reporter. The present disclosure further utilizes rapid visual detection assays capable of processing thousands of candidate microbes. The disclosed methods and biosensor can be used to identify mutant bacteria with improved nitrogen fixing capabilities. Mutant bacteria with improved nitrogen fixing capabilities are also disclosed, as well as methods of utilizing these novel bacteria to provide fixed nitrogen to a plant.
Inventors:
RYU MIN-HYUNG (US)
ESKIYENENTURK BILGE OZAYDIN (US)
TAMSIR ALVIN (US)
ESKIYENENTURK BILGE OZAYDIN (US)
TAMSIR ALVIN (US)
Application Number:
PCT/US2020/029831
Publication Date:
December 24, 2020
Filing Date:
April 24, 2020
Export Citation:
Assignee:
PIVOT BIO INC (US)
International Classes:
A01N63/00; C12N1/20; C05C1/00; C05C9/00; C12N1/21; C12N15/09; C12R1/22
Domestic Patent References:
| WO2019032926A1 | 2019-02-14 |
Other References:
AUSUBEL ET AL.: "Glutamine Synthetase Mutations Which Affect Expression of Nitrogen Fixation Genes in Klebsiella pneumoniae", J BACTERIOL, vol. 140, no. 2, 1979, pages 597 - 606, XP055773961
BENDER ET AL.: "Regulatory mutations in the Klebsiella aerogenes structural gene for glutamine synthetase", J BACTERIOL, vol. 132, no. 1, October 1977 (1977-10-01), pages 100 - 105, XP055773964
BENDER ET AL.: "Regulatory mutations in the Klebsiella aerogenes structural gene for glutamine synthetase", J BACTERIOL, vol. 132, no. 1, October 1977 (1977-10-01), pages 100 - 105, XP055773964
Attorney, Agent or Firm:
HOLLY, David Christopher et al. (US)
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