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Inactivation of phosphomannose isomerase gene abolishes sporulation and antibiotic production in Streptomyces coelicolor

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Abstract

Phosphomannose isomerases (PMIs) in bacteria and fungi catalyze the reversible conversion of d-fructose-6-phosphate to d-mannose-6-phosphate during biosynthesis of GDP-mannose, which is the main intermediate in the mannosylation of important cell wall components, glycoproteins, and certain glycolipids. In the present study, the kinetic parameters of PMI from Streptomyces coelicolor were obtained, and its function on antibiotic production and sporulation was studied. manA (SCO3025) encoding PMI in S. coelicolor was deleted by insertional inactivation. Its mutant (S. coelicolor∆manA) was found to exhibit a bld-like phenotype. Additionally, S. coelicolor∆manA failed to produce the antibiotics actinorhodin and red tripyrolle undecylprodigiosin in liquid media. To identify the function of manA, the gene was cloned and expressed in Escherichia coli BL21 (DE3). The purified recombinant ManA exhibited PMI activity (K cat/K m (mM−1 s−1 = 0.41 for d-mannose-6-phosphate), but failed to show GDP-d-mannose pyrophosphorylase [GMP (ManC)] activity. Complementation analysis with manA from S. coelicolor or E. coli resulted in the recovery of bld-like phenotype of S. coelicolor∆manA. SCO3026, another ORF that encodes a protein with sequence similarity towards bifunctional PMI and GMP, was also tested for its ability to function as an alternate ManA. However, the purified protein of SCO3026 failed to exhibit both PMI and GMP activity. The present study shows that enzymes involved in carbohydrate metabolism could control cellular differentiation as well as the production of secondary metabolites.

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Acknowledgments

This work was supported by WCU program (R322009000102130), Basic Science Research Program (2010–0009942), and Priority Research Centers Program (2009–0094021) through the National Research Foundation (NRF) grant funded by the Korean Government (MEST). This subject is also partially supported by the Korea Ministry of Environment as a “Converging Technology Project (201-101-007)” and the Converging Research Center Program through the National Research Foundation (NRF 2009-0082832).

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Authors and Affiliations

  1. Department of Microbial Engineering, College of Engineering, Konkuk University, 1 Hwayang-dong, Gwangjin-gu, Seoul, 143-701, South Korea

    Thangamani Rajesh & Yung-Hun Yang

  2. School of Chemical and Biological Engineering, Bioengineering Institute, Seoul National University, Gwanak-gu, 151-742, Seoul, South Korea

    Eunjung Song, Ji-Nu Kim, Bo-Rahm Lee, Eun-Jung Kim, Sung-Hee Park, Yun-Gon Kim, Dongwon Yoo, Yun-Hui Choi & Byung-Gee Kim

  3. Institute of Molecular Biology and Genetics, Seoul National University, Gwanak-gu, 151-742, Seoul, South Korea

    Eunjung Song, Ji-Nu Kim, Bo-Rahm Lee, Eun-Jung Kim, Sung-Hee Park, Yun-Gon Kim, Dongwon Yoo, Yun-Hui Choi & Byung-Gee Kim

  4. Bio-MAX Institute, Seoul National University, Gwanak-gu, Seoul, 151-742, South Korea

    Hyung-Yeon Park

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  1. Thangamani Rajesh
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Correspondence to Yung-Hun Yang.

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Thangamani Rajesh and Eunjung Song contributed equally to this work.

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Rajesh, T., Song, E., Kim, JN. et al. Inactivation of phosphomannose isomerase gene abolishes sporulation and antibiotic production in Streptomyces coelicolor . Appl Microbiol Biotechnol 93, 1685–1693 (2012). https://doi.org/10.1007/s00253-011-3581-z

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