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Transformable facultative thermophile Geobacillus stearothermophilus NUB3621 as a host strain for metabolic engineering

  • Applied genetics and molecular biotechnology
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Abstract

Metabolic engineers develop inexpensive enantioselective syntheses of high-value compounds, but their designs are sometimes confounded by the misfolding of heterologously expressed proteins. Geobacillus stearothermophilus NUB3621 is a readily transformable facultative thermophile. It could be used to express and properly fold proteins derived from its many mesophilic or thermophilic Bacillaceae relatives or to direct the evolution of thermophilic variants of mesophilic proteins. Moreover, its capacity for high-temperature growth should accelerate chemical transformation rates in accordance with the Arrhenius equation and reduce the risks of microbial contamination. Its tendency to sporulate in response to nutrient depletion lowers the costs of storage and transportation. Here, we present a draft genome sequence of G. stearothermophilus NUB3621 and describe inducible and constitutive expression plasmids that function in this organism. These tools will help us and others to exploit the natural advantages of this system for metabolic engineering applications.

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Acknowledgments

We thank the Bacillus Genetic Stock Center for GsNUB3621, the pNW33N plasmid, and their protocols. We also thank the Emory Integrated Genomics Core for their service and Paul Doetsch for the use of his spectrofluorimeter. This work was supported by the National Institute of General Medicine at the National Institutes for Health. KB and IM were supported by R01 GM086824 and KB was also supported by 5T32GM008490-19.

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Authors and Affiliations

  1. Department of Biochemistry, Emory University School of Medicine, 1510 Clifton Road NE, room 4119, Atlanta, GA, 30322, USA

    Kristen Blanchard & Ichiro Matsumura

  2. Department of Biology, Agnes Scott College, 141 E. College Ave., Decatur, GA, 30030, USA

    Srebrenka Robic

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  1. Kristen Blanchard
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  2. Srebrenka Robic
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Correspondence to Ichiro Matsumura.

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Blanchard, K., Robic, S. & Matsumura, I. Transformable facultative thermophile Geobacillus stearothermophilus NUB3621 as a host strain for metabolic engineering. Appl Microbiol Biotechnol 98, 6715–6723 (2014). https://doi.org/10.1007/s00253-014-5746-z

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