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Production of glycogen, PHB, biohydrogen, NAD(P)H, and proteins in Synechocystis sp. PCC 6803 disrupted in metabolically linked biosynthetic pathway(s)

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Abstract

Under nitrogen deprivation (-N), cell growth and protein synthesis of Synechocystis sp. PCC 6803 were inhibited but production of glycogen (GL) and poly-3-hydroxybutyrate (PHB) was enhanced, indicating the importance of -N for increasing the production of such bioproducts. Upon transition from N-deprived to N-supplied medium, GL and PHB were utilized for cell growth recovery. Here, we systematically disrupted the biosynthesis of GL, PHB and/or H2 and examined changes on amount of the remaining bioproducts. Disruption of PHB synthesis increased H2 evolution rate up to 1.7-fold under -N. Disruption of GL synthesis increased PHB level up to 1.4-fold, but did not affect H2 production under -N. Cellular NAD(P)H was elevated 1.6-fold after the disruption of GL synthesis, and by 3.6-fold after the disruption of both GL and PHB synthesis under -N. The double disruption of GL and PHB, GL and H2, or PHB and H2, significantly affected the initial (day 0–4) growth rate upon switching from -N to nitrogen repletion (+N). Under -N to +N condition at day 0–4, the disrupted synthesis of both GL and PHB significantly decreased the levels of total proteins, phycobilins, carotenoids, and chlorophyll a by 32%, 44%, 47%, and 59%, respectively. Thus, both PHB and GL storage are likely required for normal growth, as well as for the maximal production of proteins and photosynthetic pigments upon growth recovery under nitrogen repletion. The results demonstrated that the cyanobacterial production of GL, PHB, H2, NAD(P)H, and proteins can be affected by the disruption of metabolically connected biosynthetic pathway(s).

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Data availability

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

This work was funded by National Research Council of Thailand (NRCT): NRCT5-RSA63001-21 (to T.M.), and the Royal Golden Jubilee Ph.D. Program: PHD/0011/2560 (to J.K. and T.M.). J.K. and T.M. was supported for graduate-student research by the 90th Anniversary Fund (Ratchadaphiseksomphot Endowment Fund) of Chulalongkorn University grant (GCUGR1125633052D). The authors thank Dr. Robert Butcher for critical proofreading.

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Authors and Affiliations

  1. Department of Biochemistry, Faculty of Science, Chulalongkorn University, Bangkok, 10330, Thailand

    Janine Kaewbai-ngam, Nannaphat Sukkasam, Onuma Phoraksa, Aran Incharoensakdi & Tanakarn Monshupanee

  2. Program in Biotechnology, Faculty of Science, Chulalongkorn University, Bangkok, 10330, Thailand

    Janine Kaewbai-ngam & Nannaphat Sukkasam

  3. Academy of Science, Royal Society of Thailand, Bangkok, 10300, Thailand

    Aran Incharoensakdi

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  1. Janine Kaewbai-ngam
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Contributions

J.K. and T.M. developed the research questions and designed the experiments with the input from A.I.; J.K. mainly conducted the experiments; O.P., N.S., and T.M. generated the ΔG and ΔH strains; J.K. and T.M. analyzed the data and wrote the manuscript. All authors reviewed the manuscript.

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Correspondence to Tanakarn Monshupanee.

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Kaewbai-ngam, J., Sukkasam, N., Phoraksa, O. et al. Production of glycogen, PHB, biohydrogen, NAD(P)H, and proteins in Synechocystis sp. PCC 6803 disrupted in metabolically linked biosynthetic pathway(s). J Appl Phycol 34, 1983–1995 (2022). https://doi.org/10.1007/s10811-022-02759-2

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