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Recombinant Candida utilis for the production of biotin

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Abstract

Biotin is an important nutritional supplement but is difficult to manufacture effectively. Here we present a trial of biotin production using the food yeast Candida utilis. In this system, we cloned the C. utilis biotin synthase (BIO2) gene, the gene of the rate-limiting enzyme for biotin biosynthesis, and assembled it under the control of a strong promoter. A series of plasmids were constructed to direct the integration of the BIO2 gene, either high-copy integration with 18S rDNA fragment or low-copy integration with URA3 or HIS3 fragment. The BIO2 gene can be successfully integrated into the C. utilis chromosome and can drive biotin production using these plasmids. The biotin yield in this system can reach 100-fold above the endogenous level in a small-scale culture. Although the biotin production is not stable if the selection pressure is removed, this system has the potential to produce biotin-rich feed or food additives directly without the requirement of further purification.

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Acknowledgements

This work was partially supported by the research grants from National Science Council ROC (NSC89-2317-B110-001, NSC89-2622-B110-001, and NSC92-2311-B259-003). We also thank Professor Kuo-Chih Lin for reading the manuscript and for many valuable suggestions and the Taiwan Sugar Research Institute of Taiwan Sugar Company for their support of the work through the Academy-Enterprise Cooperation Project managed by the National Science Council.

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Correspondence to David Shiuan.

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Hong, Y., Chen, Y., Farh, L. et al. Recombinant Candida utilis for the production of biotin. Appl Microbiol Biotechnol 71, 211–221 (2006). https://doi.org/10.1007/s00253-005-0133-4

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Keywords

  • Biotin
  • Integrative Plasmid
  • Southern Hybridization Analysis
  • Pimelic Acid
  • Biotin Level