	US 12,084,705 B2
	Methylotrophic microorganisms expressing soluble methane monooxygenase proteins
Jonathan Matthew Galazka, Mountain View, CA (US); Asif Rahman, San Francisco, CA (US); and Samantha Therese Fleury, Littleton, MA (US)
Assigned to United States of America as Represented by the Administrator of NASA, Washington, DC (US)
Filed by United States of America as Represented by the Administrator of the NASA, Washington, DC (US)
Filed on Apr. 4, 2019, as Appl. No. 16/375,824.
Claims priority of provisional application 62/652,834, filed on Apr. 4, 2018.
Prior Publication US 2020/0095611 A1, Mar. 26, 2020
Int. Cl. C12N 9/04 (2006.01); C12N 1/16 (2006.01); C12N 9/02 (2006.01); C12N 15/70 (2006.01); C12P 7/00 (2006.01); C12P 7/04 (2006.01)

CPC C12P 7/04 (2013.01) [C12N 1/16 (2013.01); C12N 9/0071 (2013.01); C12Y 114/13025 (2013.01)]

19 Claims

1. An engineered methylotrophic yeast, which is a strain of Pichia pastoris, transformed to comprise:

three methane monooxygenase hydroxylase (MMOH) protein subunits of a soluble methane monooxygenase (MMO) of a methanotrophic bacterium, the three MMOH protein subunits comprising MMOH alpha, MMOH beta and MMOH gamma, and

a methane monooxygenase reductase (MMOR) of the methanotrophic bacterium,

wherein the methylotrophic yeast exhibits oxidation of methane to methanol, and

wherein the three MMOH protein subunits are expressed from a DNA construct of SEQ ID NO:42, SEQ ID NO:43, or from a DNA construct having at least 95% sequence identity to either SEQ ID NO:42, or SEQ ID NO:43.