A Novel Regulated Hybrid Promoter That Permits Autoinduction of Heterologous Protein Expression in Kluyveromyces lactis

Author:

Sakhtah Hassan1,Behler Juliane12,Ali-Reynolds Alana1,Causey Thomas B.1,Vainauskas Saulius1,Taron Christopher H.1

Affiliation:

1. New England Biolabs, Ipswich, Massachusetts, USA

2. Genetics & Experimental Bioinformatics, Institute of Biology III, University of Freiburg, Freiburg, Germany

Abstract

The yeast Kluyveromyces lactis is an important host for the expression of recombinant proteins at both laboratory and industrial scales. However, the system lacks a tightly regulated promoter that permits controlled expression of heterologous proteins. In this study, we report the engineering of a highly regulated strong hybrid promoter (termed P 350 ) for use in K. lactis . P 350 is tightly repressed by glucose or glycerol in the medium but strongly promotes gene expression once the carbon source has been consumed by the cells. This feature permits heterologous protein expression to be “autoinduced” at any scale without the addition of a gratuitous inducer molecule or changing feed solutions.

Publisher

American Society for Microbiology

Subject

Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology

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