Methods and Compositions for Targeting Heterologous Integral Membrane Proteins to the Cyanobacterial Plasma Membrane
AssigneeJoule Unlimited Technologies, Inc.
InventorNikos B. Reppas
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Abstract
This disclosure pertains to the functional localization of heterologous integral plasma membrane proteins (HIPMPs) lacking cleavable signal sequences into the plasma membrane (PM) of cyanobacterial hosts, e.g., JCC138 (Synechococcus sp. PCC 7002) or an engineered derivative thereof. More specifically, the disclosure provides chimeric integral plasma membrane proteins comprising pseudo leader sequences (PLSs) that promote increased hydrocarbon (e.g., alkane) export capabilities when expressed in a photosynthetic organism, e.g., a cyanobacterium.
Claims17
A method for modifying a heterologous integral plasma membrane protein (HIPMP) to improve its functionality in a target cyanobacterial cell, wherein said method comprises:
- (i) fusing a pseudo leader sequence (PLS) to the N-terminus of said HIPMP, wherein said HIPMP has, in its native state, its N-terminus within the cytoplasm, and wherein said PLS consists of two transmembrane alpha helices and a single periplasmic loop sequence linking the two transmembrane alpha helices;
- or
- (ii) adding a PLS to the N-terminus of said HIPMP, wherein said HIPMP has, in its native state, its N-terminus within the periplasm, and wherein said PLS consists of a single transmembrane alpha helix.
A chimeric integral plasma membrane protein (CIPMP) for facilitating hydrocarbon efflux by a target photosynthetic microorganism, wherein said CIPMP comprises, at its N-terminus, a pseudo leader sequence, wherein said pseudo leader sequence is covalently fused to a heterologous integral plasma membrane protein (IPMP), and wherein said pseudo leader sequence comprises at least one but no more than two transmembrane alpha helices, and wherein the N-terminus of said CIPMP is in the cytoplasm when expressed in said target photosynthetic microorganism.
Dependencies are inferred from the claim text.