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Caffeic acid production by simultaneous saccharification and fermentation of kraft pulp using recombinant Escherichia coli

  • Biotechnological products and process engineering
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Abstract

Caffeic acid (3,4-dihydroxycinnamic acid) serves as a building block for thermoplastics and a precursor for biologically active compounds and was recently produced from glucose by microbial fermentation. To produce caffeic acid from inedible cellulose, separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF) reactions were compared using kraft pulp as lignocellulosic feedstock. Here, a tyrosine-overproducing Escherichia coli strain was metabolically engineered to produce caffeic acid from glucose by introducing the genes encoding a 4-hydroxyphenyllactate 3-hydroxylase (hpaBC) from Pseudomonas aeruginosa and tyrosine ammonia lyase (fevV) from Streptomyces sp. WK-5344. Using the resulting recombinant strain, the maximum yield of caffeic acid in SSF (233 mg/L) far exceeded that by SHF (37.9 mg/L). In the SSF with low cellulase loads (≤2.5 filter paper unit/g glucan), caffeic acid production was markedly increased, while almost no glucose accumulation was detected, indicating that the E. coli cells experienced glucose limitation in this culture condition. Caffeic acid yield was also negatively correlated with the glucose concentration in the fermentation medium. In SHF, the formation of by-product acetate and the accumulation of potential fermentation inhibitors increased significantly with kraft pulp hydrolysate than filter paper hydrolysate. The combination of these inhibitors had synergistic effects on caffeic acid fermentation at low concentrations. With lower loads of cellulase in SSF, less potential fermentation inhibitors (furfural, 5-hydroxymethyfurfural, and 4-hydroxylbenzoic acid) accumulated in the medium. These observations suggest that glucose limitation in SSF is crucial for improving caffeic acid yield, owing to reduced by-product formation and fermentation inhibitor accumulation.

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Acknowledgments

We gratefully acknowledge Nippon Paper Group (Japan) for providing kraft pulp samples and Kouji Uematsu and Teruaki Iwanari for providing technical assistance. This work was partly supported by JST CREST Grant Number JPMJCR13B3, Japan, and by Special Coordination Funds for Promoting Science and Technology, Creation of Innovation Centers for Advanced Inter-disciplinary Research Areas (Innovative Bioproduction, Kobe), from the Ministry of Education, Culture, Sports, Science and Technology, Japan.

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Authors and Affiliations

  1. Graduate School of Science, Technology and Innovation, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501, Japan

    Hideo Kawaguchi, Hiroshi Teramura, Keiko Wakai, Kumiko Yoshihara & Ahikiko Kondo

  2. Department of Biotechnology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1, Yayoi, Bunkyo, Tokyo, 113-8657, Japan

    Yohei Katsuyama, Du Danyao & Yasuo Ohnishi

  3. Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501, Japan

    Prihardi Kahar, Sachiko Nakamura-Tsuruta & Chiaki Ogino

  4. Research Institute for Bioresources and Biotechnology, Ishikawa Prefectural University, 1-308, Suematsu, Nonoichi, Ishikawa, 921-8836, Japan

    Hiromichi Minami

  5. Biomass Engineering Research Division, RIKEN, 1-7-22 Suehiro, Turumi, Yokohama, Kanagawa, 230-0045, Japan

    Ahikiko Kondo

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  1. Hideo Kawaguchi
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  2. Yohei Katsuyama
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Correspondence to Chiaki Ogino.

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Kawaguchi, H., Katsuyama, Y., Danyao, D. et al. Caffeic acid production by simultaneous saccharification and fermentation of kraft pulp using recombinant Escherichia coli . Appl Microbiol Biotechnol 101, 5279–5290 (2017). https://doi.org/10.1007/s00253-017-8270-0

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