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Regulating Pyruvate Carboxylase in the Living Culture of Aspergillus Terreus Nrrl 1960 by l-Aspartate for Enhanced Itaconic Acid Production

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Abstract

Aspergillus terreus was reported as the promising fungal strain for itaconic acid; however, the commercial production suffers from the low yield. Low production yield was claimed as the result of completing the tricarboxylic acid (TCA) cycle towards biomass synthesis while under limiting phosphate and nitrogen; TCA cycle was somewhat shunted and consequently, the metabolite fluxes move towards itaconic acid production route. By regulating enzymes in TCA cycle, it is believed that itaconic acid production can be improved. One of the key responsible enzymes involved in itaconic acid production was triggered in this study. Pyruvate carboxylase was allosterically inhibited by l-aspartate. The presence of 10 mM l-aspartate in the production medium directly repressed PC expression in the living A. terreus while the limited malate flux regulated the malate/citrate antiporters resulting in the increasing cis-aconitate decarboxylase activity to simultaneously convert cis-aconitate, citrate isomer, into itaconic acid. The transport of cis-aconitate via the antiporters induced citrate synthase and 6-phosphofructo-1-kinase activities in response to balance the fluxes of TCA intermediates. Successively, itaconic acid production yield and final concentration could be improved by 8.33 and 60.32 %, respectively, compared to those obtained from the control fermentation with the shortened lag time to produce itaconic acid during the production phase.

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Acknowledgments

This work was supported by the Ratchadapisek Somphot Endowment Fund (2014), Chulalongkorn University (CU-57-041-AM) and the National Research Council of Thailand via the annual statement of expenditure (GRB_APS_45_57_61_03). Partial financial support from Grant for International Research Integration: Chula Research Scholar, Ratchadaphiseksomphot Endowment Fund and Thailand Research Fund via the Distinguished Research Professor Grant (DPG5880003) was also highly acknowledged. Pajareeya Songserm is the recipient of the Royal Jubilee Scholarship Program, Thailand Research Fund. The supported expenditure via this program throughout her Ph.D. study is highly acknowledged.

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Authors and Affiliations

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

    Pajareeya Songserm

  2. Institute of Biotechnology and Genetic Engineering, Chulalongkorn University, Bangkok, 10330, Thailand

    Sitanan Thitiprasert, Vasana Tolieng, Jiraporn Piluk, Aphichart Karnchanatat & Nuttha Thongchul

  3. Department of Microbiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, 10330, Thailand

    Somboon Tanasupawat

  4. Department of Chemical Engineering, Faculty of Engineering, Chulalongkorn University, Bangkok, 10330, Thailand

    Sutthichai Assabumrungrat

  5. William G. Lowrie Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, OH, 43210, USA

    Shang-Tian Yang

Authors
  1. Pajareeya Songserm
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  2. Sitanan Thitiprasert
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  3. Vasana Tolieng
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  6. Sutthichai Assabumrungrat
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  7. Shang-Tian Yang
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  8. Aphichart Karnchanatat
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  9. Nuttha Thongchul
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Correspondence to Nuttha Thongchul.

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Songserm, P., Thitiprasert, S., Tolieng, V. et al. Regulating Pyruvate Carboxylase in the Living Culture of Aspergillus Terreus Nrrl 1960 by l-Aspartate for Enhanced Itaconic Acid Production. Appl Biochem Biotechnol 177, 595–609 (2015). https://doi.org/10.1007/s12010-015-1763-3

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  • DOI: https://doi.org/10.1007/s12010-015-1763-3

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