US 12,448,609 B2
Heterologous expression of short-chain monooxygenases in microorganisms
Elizabeth Jane Clarke, San Francisco, CA (US); Baolong Zhu, San Diego, CA (US); Derek Lorin Greenfield, Kensington, CA (US); Stephanie Rhianon Jones, Berkeley, CA (US); and Noah Charles Helman, El Cerrito, CA (US)
Assigned to Industrial Microbes, Inc., Emeryville, CA (US)
Filed by INDUSTRIAL MICROBES, INC., Alameda, CA (US)
Filed on Jun. 30, 2023, as Appl. No. 18/217,021.
Application 18/217,021 is a continuation of application No. 17/149,394, filed on Jan. 14, 2021, granted, now 11,692,177.
Application 17/149,394 is a continuation of application No. 15/777,158, granted, now 10,894,951, issued on Jan. 19, 2021, previously published as PCT/US2016/062623, filed on Nov. 17, 2016.
Claims priority of provisional application 62/320,725, filed on Apr. 11, 2016.
Claims priority of provisional application 62/270,039, filed on Dec. 21, 2015.
Claims priority of provisional application 62/257,061, filed on Nov. 18, 2015.
Prior Publication US 2024/0043816 A1, Feb. 8, 2024
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 9/02 (2006.01); C12P 7/04 (2006.01); C12P 7/06 (2006.01); C12P 7/22 (2006.01)
CPC C12N 9/0073 (2013.01) [C12N 9/0071 (2013.01); C12P 7/04 (2013.01); C12P 7/06 (2013.01); C12P 7/22 (2013.01); C12Y 114/13025 (2013.01)] 13 Claims
 
1. A method for producing diiron monooxygenase, comprising:
a) culturing a synthetic microorganism comprising:
(i) at least one synthetic polynucleotide comprising at least one monooxygenase coding region encoding a diiron monooxygenase comprising SEQ ID NO: 10, wherein the diiron monooxygenase comprises one or more subunits and
(ii) at least one protein folding chaperone coding region encoding a protein folding chaperone, each linked to at least one promoter, under suitable culture conditions and for a sufficient period of time to produce the diiron monooxygenase;
b) lysing the cells, and
optionally, purifying the diiron monooxygenase to produce a lysate or purified diiron monooxygenase.