1887

Abstract

Difficulty overexpressing (eukaryotic) membrane proteins is generally considered as the major impediment in their structural and functional research. possesses many properties ideal for membrane protein expression. In order to investigate membrane protein expression in , we created a novel expression system by introducing Mistic, a short peptide previously identified in , into . The potential of this system was demonstrated in the overexpression of a eukaryotic membrane protein (pkjDes4) and a prokaryotic membrane protein (pkjLi), a newly isolated linoleate isomerase from . The expression levels reached up to 4.4 % and 45.2 % for pkjDes4 and pkjLi, respectively, which represented an exceptionally robust ability to overproduce membrane proteins. Moreover, the expressed pkjLi was functional, with its catalysing nature characterized for the first time in this species. Up to 0.852 mg ml conjugated linoleic acid was obtained during the linoleic acid conversion catalysed by the recombinant lactococcal strains. In summary, we established a membrane protein expression system in and examined its functionality. Our results demonstrate that the Mistic chaperoning strategy can be efficiently applied to hosts and show its extraordinary capacity to facilitate the high-yield production of intractable membrane proteins.

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2013-06-01
2022-08-18
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