Elsevier

Bioresource Technology

Volume 343, January 2022, 126138
Bioresource Technology

Effect of manganese peroxidase on the decomposition of cellulosic components: Direct cellulolytic activity and synergistic effect with cellulase

https://doi.org/10.1016/j.biortech.2021.126138Get rights and content

Highlights

  • Previously unknown MnP activity on cellulose decomposition was demonstrated.

  • MnP directly decomposed various cellulosic components to produce reducing sugar.

  • MnP-driven MnIII-acetate complex seemed to be essential for cellulolytic activity.

  • In addition, MnP boosted cellulase activity on carboxymethyl cellulose and Avicel®.

  • The results would contribute to construct economically feasible biorefinery system.

Abstract

Herein, it was unearthed that manganese peroxidase (MnP) from Phanerochaete chrysosporium, a lignin-degrading enzyme, is capable of not only directly decomposing cellulosic components but also boosting cellulase activity. MnP decomposes various cellulosic substrates (carboxymethyl cellulose, cellobiose [CMC], and Avicel®) and produces reducing sugars rather than oxidized sugars such as lactone and ketoaldolase. MnP with MnII in acetate buffer evolves the MnIII-acetate complex functioning as a strong oxidant, and the non-specificity of MnIII-acetate enables cellulose-decomposition. The catalytic mechanism was proposed by analyzing catalytic products derived from MnP-treated cellopentaose. Notably, MnP also boosts cellulase activity on CMC and Avicel®, even considering the cellulolytic activity of MnP itself. To the best of the authors’ knowledge, this is the first report demonstrating a previously unknown fungal MnP activity in cellulose-decomposition in addition to a known delignification activity. Consequently, the results provide a promising insight for further investigation of the versatility of lignin-degrading biocatalysts.

Introduction

As biorefinery has been burgeoning as an alternative to petroleum-based refineries (Lee et al., 2021, Moon et al., 2021, Oh et al., 2019), pretreatment for delignification and saccharification to obtain fermentable sugars are important topics in recent biorefineries utilizing lignocellulosic biomass as a sustainable feedstock (Lee et al., 2015, Min et al., 2015, Min et al., 2019, Regmi et al., 2020). Various pretreatments such as steam explosion and hydrothermolysis have been attempted, but these often suffer from a lack of selectivity, high energy consumption, and generation of inhibitors that hinder microbial fermentation (Sun & Cheng, 2002). Hence, biotechnological pretreatment using microorganism is fascinating owing to its superior selectivity and low energy consumption. White-rot fungi (e.g., Phanerochaete chrysosporium) are microorganisms capable of substantial lignin decay in nature and thus have been used for biotechnological pretreatment (Fang et al., 2020). Additionally, white-rot fungi have played as intriguing and noteworthy role in geology; the emergence of white-rot fungi might trigger a sharp decline in the rate of organic carbon burial at the end of the Carboniferous period (Floudas et al., 2012). Accordingly, it is essential to concentrate on the significance of biocatalysts secreted from white-rot fungi from the viewpoint of lignin, cellulose, and hemicellulose decomposition.

It is well-known that heme-containing oxidative enzymes (e.g. manganese peroxidase [E.C. 1.11.1.13] and lignin peroxidase [E.C 1.11.1.14]) originating from white-rot fungi are usually responsible for delignification in nature, whereas saccharification is performed by in concert with hydrolases including endo-glucanase (E.C. 3.2.1.4), exo-glucanase (E.C. 3.2.1.91), and β-glucosidase (E.C. 3.2.1.21). Recently, it was revealed that the newly classified lytic polysaccharide monooxygenase (LPMO) is also involved in saccharification by oxidative cleavage of recalcitrant polysaccharides (Beeson et al., 2012, Vaaje-Kolstad et al., 2010). This discovery has profoundly changed the understanding of biocatalytic saccharification (Forsberg et al., 2014). Given that oxidative reaction catalyzed by LPMO is involved in the saccharification of lignocellulosic biomass (Corrêa et al., 2019, Guo et al., 2020, Müller et al., 2018, Zhou et al., 2020), the raised question is whether nature-driven conversion of lignocellulosic biomass is closely related to not only hydrolysis but also oxidation in this study; if delignification and saccharification might be concurrently processed in nature, would lignin-degrading oxidative biocatalyst be able to catalyze the oxidative decomposition of cellulose as with LPMO? To answer to this question, it was examined whether lignolytic manganese peroxidase (MnP, E.C. 1.11.1.13) could catalyze recalcitrant polysaccharides in this study.

Section snippets

Materials

MnP from P. chrysosporium, cellulase from Trichoderma reesei ATCC26921, xylanase from Thermomyces lanuginosus, carboxymethyl cellulose (CMC, M.W. 90,000), p-nitrophenol cellobioside (pNPC), cellobiose, Avicel®, MnSO4, and xylan from beechwood were purchased from Sigma-Aldrich (St. Louis, USA). Hydrogen peroxide (H2O2) was purchased from Junsei (Tokyo, Japan). Regenerated amorphous cellulose (RAC) was prepared by treating Avicel® with 85 % phosphoric acid at 50 °C for 6 h, washed with distilled

MnP-driven catalysis decomposes cellulose and hemicellulose

To examine whether the lignin-degrading oxidative biocatalyst can catalyze cellulose decomposition as with LPMO, CMC was used as a soluble cellulosic substrate as a first step. Among ligninolytic biocatalysts such as lignin peroxidase, MnP, versatile peroxidase, and laccase, commercially available MnP (E.C. 1.11.1.13) from P. chrysosporium was chosen, because it oxidizes MnII to highly reactive MnIII, which is easily stabilized by carboxylic acid chelators (Hofrichter, 2002), thereby oxidizing

Conclusion

Herein, it was unearthed that MnP not only exhibited the cellulose-decomposing activities similar to endo-glucanase, exo-glucanase, xylanase, and β-glucosidase but also boosted cellulase activity as with cellulase-enhancing factor. This is the first report describing a previously unknown MnP activity in direct cellulose decomposition in addition to its primary delignification activity. The results described herein would lead to in-depth understanding of biocatalytic saccharification in nature

CRediT authorship contribution statement

Kyoungseon Min: Conceptualization, Data curation, Methodology, Formal analysis, Validation, Funding acquisition, Writing – original draft, Writing – review & editing. Yong Hwan Kim: Conceptualization, Supervision. Jiye Kim: Formal analysis, Validation. Yunje Kim: Conceptualization, Supervision. Gyeongtaek Gong: Data curation, Formal analysis, Validation, Funding acquisition. Youngsoon Um: Conceptualization, Supervision, Funding acquisition, Writing – original draft, Writing – review & editing.

Declaration of Competing Interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Acknowledgment

This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIT) (No. 2020R1A2C2008943). The authors also appreciate the additional support by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIT) (No. 2021R1A5A1028138) and the National Research Council of Science & Technology (NST) grant (No. CAP-20-02-KITOX).

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