US 12,344,878 B2
Artificial organelles for enzymatic cofactor reduction
Kyle A. Minor, Sea Bright, NJ (US); Carlo D. Montemagno, Sutton, MA (US); and David W. Wendell, Cincinnati, OH (US)
Assigned to Ensovi, Inc., Scituate, MA (US)
Filed by ENSOVI, INC., Scituate, MA (US)
Filed on Sep. 21, 2023, as Appl. No. 18/471,478.
Application 18/051,969 is a division of application No. 17/143,755, filed on Jan. 7, 2021, granted, now 11,535,880.
Application 18/471,478 is a continuation of application No. 18/051,969, filed on Nov. 2, 2022, granted, now 11,981,948.
Claims priority of provisional application 62/959,001, filed on Jan. 9, 2020.
Prior Publication US 2024/0068003 A1, Feb. 29, 2024
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 11/18 (2006.01); C07K 14/00 (2006.01); C07K 14/195 (2006.01); C07K 14/245 (2006.01); C07K 14/415 (2006.01); C12N 9/02 (2006.01); C12P 19/36 (2006.01); C12P 21/00 (2006.01)
CPC C12P 21/00 (2013.01) [C07K 14/00 (2013.01); C07K 14/195 (2013.01); C07K 14/245 (2013.01); C07K 14/415 (2013.01); C12N 9/0036 (2013.01); C12N 11/18 (2013.01); C12P 19/36 (2013.01); C12Y 106/05003 (2013.01)] 20 Claims
OG exemplary drawing
 
1. An artificial cell free organelle system comprising:
a membrane having two sides comprising an inner surface in contact with an inner aqueous medium and an outer surface in contact with an outer aqueous medium;
one or more photosynthetic proteins comprising a photosystem II complex of proteins or a bacteriorhodopsin vectorially embedded within and traversing the membrane;
one or more oxidoreductase proteins comprising a Respiratory Complex I complex of proteins vectorially embedded within and traversing the membrane;
one or more first redox active cofactors comprising ubiquinone or analogues thereof;
one or more second redox active cofactors comprising NAD+, NADP+ or an analogue thereof;
water, and
a photon energy source;
wherein:
when one or more photons are directed on the one or more photosynthetic proteins, the one or more photosynthetic proteins harvest energy from the one or more photons and catalyze an oxidation of at least one water molecule in the inner aqueous medium, generating 0.5 equivalents of oxygen gas and yielding up to two protons and two electrons per two photons that are transferred to an oxidized form of the one or more first redox active cofactors, generating a reduced form of the one or more first redox active cofactors;
accumulation of protons in the inner aqueous medium generates a proton concentration gradient between the inner aqueous medium and the outer aqueous medium; and
the one or more oxidoreductase proteins pump protons from the inner aqueous medium through the membrane to the outer aqueous medium to reduce the proton concentration gradient and simultaneously catalyzes the transfer of electrons from the reduced form of the one or more first redox active cofactors to an oxidized form of the one or more second redox active cofactors, generating a reduced form of the one or more second redox active cofactors and an oxidized form of the one or more first redox active cofactors.