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Identification and Biochemical Characterization of Major β-Mannanase in Talaromyces cellulolyticus Mannanolytic System

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Abstract

Talaromyces cellulolyticus is a promising fungus for providing a cellulase preparation suitable for the hydrolysis of lignocellulosic material, although its mannan-degrading activities are insufficient. In the present study, three core mannanolytic enzymes, including glycosyl hydrolase family 5–7 (GH5–7) β-mannanase (Man5A), GH27 α-galactosidase, and GH2 β-mannosidase, were purified from a culture supernatant of T. cellulolyticus grown with glucomannan, and the corresponding genes were identified based on their genomic sequences. Transcriptional analysis revealed that these genes were specifically induced by glucomannan. Two types of Man5A products, Man5A1 and Man5A2, were found as major proteins in the mannanolytic system. Man5A1 was devoid of a family 1 carbohydrate-binding module (CBM1) at the N-terminus, whereas Man5A2 was devoid of both CBM1 and Ser/Thr-rich linker region. The physicochemical and catalytic properties of both Man5A1 and Man5A2 were identical to those of recombinant Man5A (rMan5A) possessing CBM1, except for the cellulose-binding ability. Man5A CBM1 had little effect on mannan hydrolysis of pretreated Hinoki cypress. The results suggest that an improvement in Man5A CBM1 along with the augmentation of identified mannanolytic enzyme components would aid in efficient hydrolysis of softwood using T. cellulolyticus cellulase preparation.

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Acknowledgments

The authors are grateful to Dr. Akio Kumagai (National Institute of Advanced Industrial Science and Technology) for helpful discussions.

Funding

This work was financially supported by a Basic Research Funding grant from the National Institute of Advanced Industrial Science and Technology.

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Authors and Affiliations

  1. Department of Bioscience and Biotechnology, Faculty of Agriculture, University of the Ryukyus, 1 Senbaru, Nishihara, Okinawa, 903-0213, Japan

    Keiko Uechi

  2. Research Institute for Sustainable Chemistry, National Institute of Advanced Industrial Science and Technology (AIST), 3-11-32 Kagamiyama, Higashi-Hiroshima, Hiroshima, 739-0046, Japan

    Masahiro Watanabe, Tatsuya Fujii, Saori Kamachi & Hiroyuki Inoue

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  1. Keiko Uechi
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  2. Masahiro Watanabe
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  3. Tatsuya Fujii
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Contributions

All authors contributed to the study conception and design. K.U, S.K, and H.I prepared and characterized the enzymes. T.F characterized the gene expression. M.W and H.I coordinated the study. All authors contributed to the writing of this manuscript and approved the final version.

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Correspondence to Hiroyuki Inoue.

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Uechi, K., Watanabe, M., Fujii, T. et al. Identification and Biochemical Characterization of Major β-Mannanase in Talaromyces cellulolyticus Mannanolytic System. Appl Biochem Biotechnol 192, 616–631 (2020). https://doi.org/10.1007/s12010-020-03350-6

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