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Enhanced tolerance of Cupriavidus necator NCIMB 11599 to lignocellulosic derived inhibitors by inserting NAD salvage pathway genes

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Abstract

Polyhydroxybutyrate (PHB) is a bio-based, biodegradable and biocompatible plastic that has the potential to replace petroleum-based plastics. Lignocellulosic biomass is a promising feedstock for industrial fermentation to produce bioproducts such as polyhydroxybutyrate (PHB). However, the pretreatment processes of lignocellulosic biomass lead to the generation of toxic byproducts, such as furfural, 5-HMF, vanillin, and acetate, which affect microbial growth and productivity. In this study, to reduce furfural toxicity during PHB production from lignocellulosic hydrolysates, we genetically engineered Cupriavidus necator NCIMB 11599, by inserting the nicotine amide salvage pathway genes pncB and nadE to increase the NAD(P)H pool. We found that the expression of pncB was the most effective in improving tolerance to inhibitors, cell growth, PHB production and sugar consumption rate. In addition, the engineered strain harboring pncB showed higher PHB production using lignocellulosic hydrolysates than the wild-type strain. Therefore, the application of NAD salvage pathway genes improves the tolerance of Cupriavidus necator to lignocellulosic-derived inhibitors and should be used to optimize PHB production.

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Funding

This study was supported by the National Research Foundation of Korea (NRF) (NRF-2019M3E6A1103979 and NRF2022R1A2C2003138). This work was also supported by the R&D Program of MOTIE/KEIT (20015373, 20016324) and with the support of the R&D Program for Forest Science Technology [grant number 2020261C10-2022-AC02] provided by the Korea Forest Service (Korea Forestry Promotion Institute).

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Authors and Affiliations

  1. Department of Biological Engineering, College of Engineering, Konkuk University, Seoul, Republic of Korea

    Sun Mi Lee, Do-Hyun Cho, Hee Ju Jung, Byungchan Kim, Su Hyun Kim, Shashi Kant Bhatia, Ranjit Gurav & Yung-Hun Yang

  2. Green & Sustainable Materials R&D Department, Korea Institute of Industrial Technology (KITECH), Cheonan-si, Republic of Korea

    Jong-Min Jeon & Jeong-Jun Yoon

  3. Sustainable Technology and Wellness R&D Group, Korea Institute of Industrial Technology (KITECH), Jeju-si, Republic of Korea

    Jeong-Hoon Park

  4. Bio-Evaluation Center, Korea Research Institute of Bioscience and Biotechnology, Cheongju, Republic of Korea

    Jung-Ho Park

  5. Department of Chemical Engineering, Soongsil University, Seoul, Republic of Korea

    Yun-Gon Kim

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  1. Sun Mi Lee
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Contributions

SML and Y-HY conceived and designed the study; SML, D-HC, HJJ, BK, SHK performed the experiments and drafted the manuscript; SKB., RG., and Y-HY interpreted the experimental results; SML, J-MJ, J-JY, J-HP, J-HP, Y-GK and Y-HY revised the manuscript. All authors have read and approved the final manuscript.

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Correspondence to Yung-Hun Yang.

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Lee, S.M., Cho, DH., Jung, H.J. et al. Enhanced tolerance of Cupriavidus necator NCIMB 11599 to lignocellulosic derived inhibitors by inserting NAD salvage pathway genes. Bioprocess Biosyst Eng 45, 1719–1729 (2022). https://doi.org/10.1007/s00449-022-02779-9

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