US 12,410,458 B2
Semi-continuous culture methods
Zhiyong Sun, Dartmouth (CA); Michael Milway, Dartmouth (CA); Kevin Berryman, Dartmouth (CA); Mercia Valentine, Dartmouth (CA); Roberto E. Armenta, Dartmouth (CA); and Laura Purdue, Dartmouth (CA)
Assigned to Mara Renewables Corporation, Dartmouth (CA)
Filed by Mara Renewables Corporation, Dartmouth (CA)
Filed on Apr. 25, 2022, as Appl. No. 17/728,833.
Application 17/728,833 is a continuation of application No. 16/864,128, filed on Apr. 30, 2020, granted, now 11,345,943.
Application 16/864,128 is a continuation of application No. 14/882,734, filed on Oct. 14, 2015, granted, now 10,676,775, issued on Jun. 9, 2020.
Claims priority of provisional application 62/064,668, filed on Oct. 16, 2014.
Prior Publication US 2022/0356506 A1, Nov. 10, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. C12Q 1/06 (2006.01); C12N 1/04 (2006.01); C12N 1/12 (2006.01); C12N 1/14 (2006.01); C12P 7/6409 (2022.01); C12P 7/6427 (2022.01); C12P 7/6431 (2022.01); C12P 7/6432 (2022.01); C12P 7/6434 (2022.01); C12P 7/6463 (2022.01); C12P 7/6472 (2022.01)
CPC C12Q 1/06 (2013.01) [C12N 1/04 (2013.01); C12N 1/12 (2013.01); C12N 1/14 (2013.01); C12P 7/6409 (2013.01); C12P 7/6427 (2013.01); C12P 7/6431 (2022.01); C12P 7/6432 (2022.01); C12P 7/6434 (2022.01); C12P 7/6463 (2013.01); C12P 7/6472 (2013.01)] 20 Claims
 
1. A method of producing a biomass of oil-producing microorganisms comprising the steps of:
(a) culturing the oil-producing microorganisms in a container with a medium comprising a first carbon to nitrogen ratio until the culture reaches a threshold indicator, wherein the threshold indicator is the concentration of carbon in the culture medium and wherein the concentration of carbon is at or near zero;
(c) harvesting a 5% to 20% of the culture and maintaining the majority of the culture in the container; and
(d) promoting production of the biomass of oil-producing microorganisms by adding fresh medium to the container at a volume of substantially the same volume as the harvested portion, wherein the fresh medium comprises a second carbon to nitrogen ratio.