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.
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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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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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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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DOI: https://doi.org/10.1007/s12010-020-03350-6