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Heterologous gene expression in Thermus thermophilus: β-galactosidase, dibenzothiophene monooxygenase, PNB carboxy esterase, 2-aminobiphenyl-2,3-diol dioxygenase, and chloramphenicol acetyl transferase

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Journal of Industrial Microbiology and Biotechnology

Abstract

Enzymes from thermophiles are preferred for industrial applications because they generally show improved tolerance to temperature, pressure, solvents, and pH as compared with enzymes from mesophiles. However, nearly all thermostable enzymes used in industrial applications or available commercially are produced as recombinant enzymes in mesophiles, typically Escherichia coli. The development of high-temperature bioprocesses, particularly those involving cofactor-requiring enzymes and/or multi-step enzymatic pathways, requires a thermophilic host. The extreme thermophile most amenable to genetic manipulation is Thermus thermophilus, but the study of expression of heterologous genes in T. thermophilus is in its infancy. While several heterologous genes have previously been expressed in T. thermophilus (Fridjonsson et al. in J Bacteriol 184:3385–3391, 2002, Koyama et al. in Appl Environ Microbiol 56:2251–225, 1990, Lasa et al. in J Bacteriol 174:6424–6431, 1992, Mathew et al. in Appl Environ Microbiol 58:421–425, 1992, Takagi et al. in J Ind Microbiol Biotechnol 23:214–217, 1999, Tamakoshi et al. in Extremophiles 5:17–22 2001), the data reported here include the first examples of the functional expression of a gene from an archaeal hyperthermophile (bglA from Pyrococcus woesei), a cofactor-requiring enzyme (dszC from Rhodococcus erythropolis IGTS8), and a two-component enzyme (carBa and carBb from Sphingomonas sp. GTIN11). A thermostable derivative of pnbA from Bacillus subtilis was also expressed, further expanding the list of genes from heterologous hosts that have been expressed in T. thermophilus.

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Acknowledgements

This project was supported by DOE contract DE-AC26–99BC15219.

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Authors and Affiliations

  1. Department of Microbiology, University of Illinois at Urbana-Champaign, 601 S. Goodwin Ave., Urbana, IL 61801, USA

    Ho-Shin Park

  2. Sigma-Aldrich, 2909 Lacleide Ave, St. Louis, MO 63103, USA

    Kevin J. Kayser

  3. Department of Biochemistry, University of Washington, Box 357350, Seattle, WA 98195-7350, USA

    Jung-Ho Kwak

  4. Gas Technology Institute, 1700 S. Mt. Prospect Road, Des Plaines, IL 60018, USA

    John J. Kilbane II

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  1. Ho-Shin Park
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Correspondence to John J. Kilbane II.

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Park, HS., Kayser, K.J., Kwak, JH. et al. Heterologous gene expression in Thermus thermophilus: β-galactosidase, dibenzothiophene monooxygenase, PNB carboxy esterase, 2-aminobiphenyl-2,3-diol dioxygenase, and chloramphenicol acetyl transferase. J IND MICROBIOL BIOTECHNOL 31, 189–197 (2004). https://doi.org/10.1007/s10295-004-0130-0

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