Abstract
(2S, 3R)-Hydroxycitric acid (HCA) from Hibiscus subdariffa inhibits pancreatic α-amylase and intestine α-glucosidase, leading to reduction of carbohydrate metabolism. In our previous study, Streptomyces sp. U121 was identified as a producer of (2S, 3R)-HCA [Hida et al. (2005) Bioscience, Biotechnology, and Biochemistry 69:1555–1561]. Here, we applied genome shuffling of Streptomyces sp. U121 to achieve rapid improvement of HCA production. The initial mutant population was generated by nitrosoguanidine treatment of the spores, and an improved population producing fivefold more HCA over wild type was obtained by three rounds of genome shuffling. For efficient screening of the mutant library, trans-epoxyaconitic acid (EAA), an antibiotic analog of HCA, was utilized. EAA inhibited the regeneration of nonfused protoplasts, resulting in selective screening of shuffled strains. Mutant strains with enhanced EAA resistance exhibited significantly higher HCA production in liquid media. Furthermore, the best mutant showed increased cell growth in flask culture, as well as increased HCA production.
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Acknowledgments
We thank Y. Nakaoka and M. Tokugawa for screening of the mutant library. This work was in part supported by the “High-Tech Research Center” Project for Private Universities on Evolution of Green Science for Quality Improvement of Environmental and Health: matching fund subsidy from the Ministry of Education, Culture, Sports, Science and Technology, 2004–2008.
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Hida, H., Yamada, T. & Yamada, Y. Genome shuffling of Streptomyces sp. U121 for improved production of hydroxycitric acid. Appl Microbiol Biotechnol 73, 1387–1393 (2007). https://doi.org/10.1007/s00253-006-0613-1
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DOI: https://doi.org/10.1007/s00253-006-0613-1