Genetic Toolbox for Controlled Expression of Functional Proteins in Geobacillus Spp

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2017 Feb 3

PMID 28152017

Citations 11

Authors

Ivan Pogrebnyakov

Christian Bille Jendresen

Alex Toftgaard Nielsen

Affiliations

Soon will be listed here.

Abstract

Species of genus Geobacillus are thermophilic bacteria and play an ever increasing role as hosts for biotechnological applications both in academia and industry. Here we screened a number of Geobacillus strains to determine which industrially relevant carbon sources they can utilize. One of the strains, G. thermoglucosidasius C56-YS93, was then chosen to develop a toolbox for controlled gene expression over a wide range of levels. It includes a library of semi-synthetic constitutive promoters (76-fold difference in expression levels) and an inducible promoter from the xylA gene. A library of synthetic in silico designed ribosome binding sites was also created for further tuning of translation. The PxylA was further used to successfully express native and heterologous xylanases in G. thermoglucosidasius. This toolbox enables fine-tuning of gene expression in Geobacillus species for metabolic engineering approaches in production of biochemicals and heterologous proteins.

Citing Articles

Development of a thermophilic host-vector system for the production of recombinant proteins at elevated temperatures.

Kurashiki R, Koyama K, Sakaguchi Y, Okumura Y, Ohshiro T, Suzuki H Appl Microbiol Biotechnol. 2023; 107(24):7475-7488.

PMID: 37755510 DOI: 10.1007/s00253-023-12805-9.

An improved integrative GFP-based vector for genetic engineering of Parageobacillus thermoglucosidasius facilitates the identification of a key sporulation regulator.

Millgaard M, Bidart G, Pogrebnyakov I, Nielsen A, Welner D AMB Express. 2023; 13(1):44.

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New Platform for Screening Genetic Libraries at Elevated Temperatures: Biological and Genomic Information and Genetic Tools of Geobacillus thermodenitrificans K1041.

Koyama K, Mikawa Y, Nakagawa S, Kurashiki R, Ohshiro T, Suzuki H Appl Environ Microbiol. 2022; 88(18):e0105122.

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Establishment of a functional system for recombinant production of secreted proteins at 50 °C in the thermophilic Bacillus methanolicus.

Irla M, Drejer E, Brautaset T, Hakvag S Microb Cell Fact. 2020; 19(1):151.

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A plasmid vector that directs hyperproduction of recombinant proteins in the thermophiles Geobacillus species.

Kurashiki R, Mizuno T, Murata K, Ohshiro T, Suzuki H Extremophiles. 2019; 24(1):147-156.

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