Identification and enzymatic characterization of an endo-1,3-β-glucanase from Euglena gracilis
Abstract
Euglena produces paramylon as a storage polysaccharide, and is thought to require β-1,3-glucan degrading enzymes to release and utilize the accumulated carbohydrate. To investigate β-1,3-glucan degradation in Euglena, endo-1,3-β-glucanases were partially purified from Euglena gracilis by hydrophobic, gel filtration and anion-exchange chromatography. Tryptic digests and mass-spectrometric analysis identified three proteins in the purified fraction as a member of glycoside hydrolase family (GH) 17 and two members of GH81. These genes were cloned from an Euglena cDNA pool by PCR. EgCel17A fused with a histidine-tag at the carboxy terminus was heterologously produced by Aspergillus oryzae and purified by immobilized metal affinity chromatography. Purified EgCel17A had a molecular weight of about 40kDa by SDS-PAGE, which was identical to that deduced from its amino acid sequence. The enzyme showed hydrolytic activity towards β-1,3-glucans such as laminarin and paramylon. Maximum activity of laminarin degradation by EgCel17A was attained at pH 4.0-5.5 and 60°C after 1h incubation or 50°C after 20h incubation. The enzyme had a Km of 0.21mg/ml and a Vmax of 40.5units/mg protein for laminarin degradation at pH 5.0 and 50°C. Furthermore, EgCel17A catalyzed a transglycosylation reaction by which reaction products with a higher molecular weight than the supplied substrates were initially generated; however, ultimately the substrates were degraded into glucose, laminaribiose and laminaritriose. EgCel17A effectively produced soluble β-1,3-glucans from alkaline-treated Euglena freeze-dried powder containing paramylon. Thus, EgCel17 is the first functional endo-1,3-β-glucanase to be identified from E. gracilis.
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Degradation of paramylon to glucose is simpler than laminarin and requires the same enzymatic arsenal, with the exception of the debranching endo-β-(1→6) glucanases.
“Therefore, degradation of paramylon to glucose is simpler than laminarin and requires the same enzymatic arsenal, with the exception of the debranching endo-β-(1→6) glucanases (Takeda et al., 2015).”
From The Enzymatic Conversion of Major Algal and Cyanobacterial Carbohydrates to Bioethanol (2016) · cites this paper as Takeda et al., 2015
Cites
A classification of glycosyl hydrolases based on amino acid sequence similarities.
Henrissat, B. (1991) · Biochemical Journal · doi:10.1042/bj2800309 · Cited by 3,188 · see citing works
Structural and sequence-based classification of glycoside hydrolases
Henrissat, B., & Davies, G. J. (1997) · Current Opinion in Structural Biology · doi:10.1016/s0959-440x(97)80072-3 · Cited by 1,668 · see citing works
Updating the sequence-based classification of glycosyl hydrolases
Henrissat, B., & Bairoch, A. (1996) · Biochemical Journal · doi:10.1042/bj3160695 · Cited by 1,362 · see citing works
A new reaction for colorimetric determination of carbohydrates
Lever, M. (1972) · Analytical Biochemistry · doi:10.1016/0003-2697(72)90301-6 · Cited by 900 · see citing works
Xyloglucan endotransglycosylase, a new wall-loosening enzyme activity from plants
Fry, S. C., Smith, R. C., Renwick, K. F., Martin, D. J., Hodge, S., & Matthews, K. J. (1992) · Biochemical Journal · doi:10.1042/bj2820821 · Cited by 833 · see citing works
McCormick, S. P. (1993) · The Plant Cell · doi:10.1105/tpc.5.10.1265 · Cited by 517 · see citing works
Microbial β-Glucosidases: Cloning, Properties, and Applications
Bhatia, Y., Mishra, S., & Bisaria, V. S. (2002) · Critical Reviews in Biotechnology · doi:10.1080/07388550290789568 · Cited by 498 · see citing works
Nishitani, K., & Tominaga, R. (1992) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(19)36797-3 · Cited by 496 · see citing works
Callose synthesis in higher plants
Chen, X., & Kim, J. (2009) · Plant Signaling & Behavior · doi:10.4161/psb.4.6.8359 · Cited by 479 · see citing works
Gomi, K., Iimura, Y., & Hara, S. (1987) · Agricultural and Biological Chemistry · doi:10.1271/bbb1961.51.2549 · Cited by 307 · see citing works
A Cell Plate-Specific Callose Synthase and Its Interaction with Phragmoplastin
Hong, Z., Delauney, A. J., & Verma, D. P. S. (2001) · The Plant Cell · doi:10.1105/tpc.13.4.755 · Cited by 288 · see citing works
Genome-wide identification of fungal GPI proteins.
de Groot, P. W. J., Hellingwerf, K. J., & Klis, F. M. (2003) · Yeast · doi:10.1002/yea.1007 · Cited by 276 · see citing works
Kawamura, Y., & Uemura, M. (2003) · The Plant Journal · doi:10.1046/j.1365-313x.2003.01864.x · Cited by 269 · see citing works
McCormick, S. M. (1993) · The Plant Cell · doi:10.2307/3869779 · Cited by 238 · see citing works
Michel, G., Tonon, T., Scornet, D., Cock, J. M., & Kloareg, B. (2010) · New Phytologist · doi:10.1111/j.1469-8137.2010.03345.x · Cited by 188 · see citing works
Ruel, K., & Joseleau, J. P. (1991) · Applied and Environmental Microbiology · doi:10.1128/aem.57.2.374-384.1991 · Cited by 131 · see citing works
A Novel β-( , , )-Glucanosyltransferase from the Cell Wall of Aspergillus fumigatus
Hartland, R. P., Fontaine, T., Debeaupuis, J. P., Simenel, C., Delepierre, M., & Latgé, J. (1996) · Journal of Biological Chemistry · doi:10.1074/jbc.271.43.26843 · Cited by 117 · see citing works
Sietsma, J. H., & Wessels, J. G. H. (1981) · Microbiology · doi:10.1099/00221287-125-1-209 · Cited by 106 · see citing works
van Bueren, A. L., Morland, C., Gilbert, H. J., & Boraston, A. B. (2004) · Journal of Biological Chemistry · doi:10.1074/jbc.m410113200 · Cited by 100 · see citing works
High yield media for photosynthesizing euglena gracilis abstract
Koren, L. E., & Hutner, S. H. (1967) · Journal of Eukaryotic Microbiology · Cited by 76 · see citing works
Screening of various glycosidases for the synthesis of octyl glucoside
Ducret, A., Trani, M., & Lortie, R. (2002) · Biotechnology and Bioengineering · doi:10.1002/bit.10156 · Cited by 63 · see citing works
Cline, K., & Albersheim, P. (1981) · PLANT PHYSIOLOGY · doi:10.1104/pp.68.1.207 · Cited by 54 · see citing works
Peumans, W. J., Barre, A., Derycke, V., Rougé, P., Zhang, W., May, G. D., Delcour, J. A., LEUVEN, F. V., & Damme, E. J. M. V. (2000) · European Journal of Biochemistry · doi:10.1046/j.1432-1327.2000.01117.x · Cited by 53 · see citing works
β-1,3-glucan hydrolases from Euglena gracilis
Barras, D. R., & Stone, B. A. (1969) · Biochimica et Biophysica Acta (BBA) - Enzymology · doi:10.1016/0005-2744(69)90253-8 · Cited by 53 · see citing works
β1,3-Glucan hydrolases from Euglena gracilis
Barras, D. R., & Stone, B. A. (1969) · Biochimica et Biophysica Acta (BBA) - Enzymology · doi:10.1016/0005-2744(69)90252-6 · Cited by 47 · see citing works
Johnston, S. L., Prakash, R., Chen, N. J., Kumagai, M. H., Turano, H., Cooney, J. M., Atkinson, R. G., Paull, R. E., Cheetamun, R., Bacic, A., et al. (2012) · Planta · doi:10.1007/s00425-012-1766-z · Cited by 40 · see citing works
Gupta, R., Govil, T., Capalash, N., & Sharma, P. (2012) · Applied Biochemistry and Biotechnology · doi:10.1007/s12010-012-9889-z · Cited by 36 · see citing works
Cheng, R., Chen, J., Yu, X., Wang, Y., Wang, S., & Zhang, J. (2013) · BMC Biotechnology · doi:10.1186/1472-6750-13-105 · Cited by 34 · see citing works
Secretory expression of the non-secretory-type Lentinula edodes laccase by Aspergillus oryzae
Yano, A., Kikuchi, S., Nakagawa, Y. S., Sakamoto, Y., & Sato, T. (2009) · Microbiological Research · doi:10.1016/j.micres.2008.12.001 · Cited by 31 · see citing works
Biochemical characterization of Magnaporthe oryzae β-glucosidases for efficient β-glucan hydrolysis
Takahashi, M., Konishi, T., & Takeda, T. (2011) · Applied Microbiology and Biotechnology · doi:10.1007/s00253-011-3340-1 · Cited by 24 · see citing works
Cloning and characterization of a new β-mannosidase from Streptomyces sp. S27
Shi, P., Yao, G., Cao, Y., Yang, P., Yuan, T., Huang, H., Bai, Y., & Yao, B. (2011) · Enzyme and Microbial Technology · doi:10.1016/j.enzmictec.2011.06.003 · Cited by 24 · see citing works
Nakajima, M., Yamashita, T., Takahashi, M., Nakano, Y., & Takeda, T. (2012) · Applied and Environmental Microbiology · doi:10.1128/aem.00483-12 · Cited by 20 · see citing works
Ammonia- and Light-Induced Degradation of Paramylum in Euglena gracilis
Sumida, S., Ehara, T., Osafune, T., & Hase, E. (1987) · Plant and Cell Physiology · doi:10.1093/oxfordjournals.pcp.a077454 · Cited by 16 · see citing works
Takahashi, M., Yoshioka, K., Imai, T., Miyoshi, Y., Nakano, Y., Yoshida, K., Yamashita, T., Furuta, Y., Watanabe, T., Sugiyama, J., et al. (2013) · Journal of Biological Chemistry · doi:10.1074/jbc.m112.448902 · Cited by 11 · see citing works
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Cited by
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Microalgae and cyanobacteria as enzyme biofactories
Brasil, B. S. A. F., de Siqueira, F. G., Salum, T. F. C., Zanette, C. M., & Spier, M. R. (2017) · Algal Research · doi:10.1016/j.algal.2017.04.035 · Cited by 199 · see citing works
Bioproducts From Euglena gracilis: Synthesis and Applications
Gissibl, A., Sun, A., Care, A., Nevalainen, H. K. M., & Sunna, A. (2019) · Frontiers in Bioengineering and Biotechnology · doi:10.3389/fbioe.2019.00108 · Cited by 199 · see citing works
Yoshida, Y., Tomiyama, T., Maruta, T., Tomita, M., Ishikawa, T., & Arakawa, K. (2016) · BMC Genomics · doi:10.1186/s12864-016-2540-6 · Cited by 103 · see citing works
The Enzymatic Conversion of Major Algal and Cyanobacterial Carbohydrates to Bioethanol
Al Abdallah, Q., Nixon, B. T., & Fortwendel, J. R. (2016) · Frontiers in Energy Research · doi:10.3389/fenrg.2016.00036 · Cited by 102 · see citing works
Huang, Y., Jiang, H., Mao, X., & Ci, F. (2021) · Journal of Ocean University of China · doi:10.1007/s11802-021-4584-8 · Cited by 58 · see citing works
Wax Ester Fermentation and Its Application for Biofuel Production
Inui, H., Ishikawa, T., & Tamoi, M. (2017) · Advances in experimental medicine and biology · doi:10.1007/978-3-319-54910-1_13 · Cited by 58 · see citing works
Xu, J., Zhao, Y., Zhang, X., Zhang, L., Hou, Y., & Dong, W. (2016) · Frontiers in Plant Science · doi:10.3389/fpls.2016.01524 · Cited by 45 · see citing works
Kuhaudomlarp, S., Patron, N. J., Henrissat, B., Rejzek, M., Saalbach, G., & Field, R. A. (2018) · Journal of Biological Chemistry · doi:10.1074/jbc.ra117.000936 · Cited by 45 · see citing works
Laminarans and 1,3-β-D-glucanases
Usoltseva, R. V., Belik, A. A., Игоревич, К. М., Malyarenko, O. S., Николаевна, З. Т., & Ermakova, S. P. (2020) · International Journal of Biological Macromolecules · doi:10.1016/j.ijbiomac.2020.07.034 · Cited by 45 · see citing works
Euglena Central Metabolic Pathways and Their Subcellular Locations
Inwongwan, S., Kruger, N. J., Ratcliffe, R. G., & O’Neill, E. C. (2019) · Metabolites · doi:10.3390/metabo9060115 · Cited by 33 · see citing works
Cell wall structure of secreted laccase-silenced strain in Lentinula edodes
Sakamoto, Y., Nakade, K., Sato, S., Yoshimi, A., Sasaki, K., Konno, N., & Abe, K. (2018) · Fungal Biology · doi:10.1016/j.funbio.2018.09.005 · Cited by 32 · see citing works
Expression and Characterization of a Novel Antifungal Exo-β-1,3-glucanase from Chaetomium cupreum
Jiang, C., Song, J., Cong, H., Zhang, J., & Yang, Q. (2016) · Applied Biochemistry and Biotechnology · doi:10.1007/s12010-016-2325-z · Cited by 32 · see citing works
Wang, D., Kim, D. H., Seo, N., Yun, E. J., An, H. J., Kim, J., & Kim, K. H. (2016) · Applied and Environmental Microbiology · doi:10.1128/aem.00635-16 · Cited by 28 · see citing works
Yang, Y., Guo, J., Cheng, J., Jiang, Z., Xu, N., An, X., Chen, Z., Hao, J., Yang, S., Xu, Z., et al. (2020) · Industrial Crops and Products · doi:10.1016/j.indcrop.2020.112484 · Cited by 24 · see citing works
Gissibl, A., Care, A., Parker, L. M., Iqbal, S., Hobba, G. D., Nevalainen, H. K. M., & Sunna, A. (2018) · Carbohydrate Polymers · doi:10.1016/j.carbpol.2018.05.038 · Cited by 23 · see citing works
Takeda, T., Takahashi, M., Shimizu, M., Sugihara, Y., Yamashita, T., Saitoh, H., Fujisaki, K., Ishikawa, K., Utsushi, H., Kanzaki, E., et al. (2022) · PLoS Pathogens · doi:10.1371/journal.ppat.1010792 · Cited by 22 · see citing works
Xu, J., Zhang, Y., Qi, D., Huo, H., Dong, X., Tian, L., Liu, C., & Cao, Y. (2020) · Genomics · doi:10.1016/j.ygeno.2020.10.025 · Cited by 22 · see citing works
Advances in molecular enzymology of β-1,3-glucanases: A comprehensive review.
Jiang, Y., Chang, Z., Xu, Y., Zhan, X.-B., Wang, Y.-Y., & Gao, M. (2024) · International Journal of Biological Macromolecules · doi:10.1016/j.ijbiomac.2024.135349 · Cited by 22 · see citing works
Wang, Y., Li, D., Dong, C., Zhao, Y., Zhang, L., Yang, F., Ye, X., Huang, Y., Li, Z., & Cui, Z. (2021) · Applied Microbiology and Biotechnology · doi:10.1007/s00253-021-11513-6 · Cited by 12 · see citing works
Zhang, Z., Yu, S., Gu, J., Tan, F., Wan, S., Gao, H., & Qin, Z. (2024) · Journal of Agricultural and Food Chemistry · doi:10.1021/acs.jafc.4c08008 · Cited by 10 · see citing works
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Li, Z., Liu, W., & Lyu, Q. (2020) · Marine Drugs · doi:10.3390/md18090466 · Cited by 10 · see citing works
Abi, A., Müller, C., & Jördening, H. (2017) · Biotechnology and Bioprocess Engineering · doi:10.1007/s12257-016-0649-8 · Cited by 10 · see citing works
Jiang, C., Miao, G., Li, J., Zhang, Z., Li, J., Zhu, S., Zhang, J., & Zhou, X. (2023) · Applied Biochemistry and Biotechnology · doi:10.1007/s12010-023-04698-1 · Cited by 9 · see citing works
Zamora‐Carreras, H., Torres, M., Bustamante, N., Macedo, A. L., Rodrı́guez, R., Villalba, M., & Bruix, M. (2015) · Archives of Biochemistry and Biophysics · doi:10.1016/j.abb.2015.07.004 · Cited by 8 · see citing works
Tanaka, Y., Goto, K., Luo, J., Nishino, K., Ogawa, T., Maruta, T., & Ishikawa, T. (2022) · Algal Research · doi:10.1016/j.algal.2022.102829 · Cited by 6 · see citing works
Functional and structural characterization of an endo-β-1,3-glucanase from Euglena gracilis
Calloni, R. D., Muchut, R. J., Garay, A. S., Arias, D. G., Iglesias, A. Á., & Guerrero, S. A. (2022) · Biochimie · doi:10.1016/j.biochi.2022.12.016 · Cited by 5 · see citing works
Ebenezer, T. E., Zoltner, M., Burrell, A., Nenarokova, A., Vanclová, A. M. G. N., Prasad, B., Soukal, P., Santana‐Molina, C., O’Neill, E. C., Nankissoor, N. N., et al. (2017) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/228015 · Cited by 4 · see citing works
Development of screening strategies for the identification of paramylon-degrading enzymes
Gissibl, A., Care, A., Sun, A., Hobba, G. D., Nevalainen, H. K. M., & Sunna, A. (2019) · Journal of Industrial Microbiology & Biotechnology · doi:10.1007/s10295-019-02157-7 · Cited by 2 · see citing works
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Baylan, M., & Gamze, M. A. Z. I. (2022) · Osmaniye Korkut Ata Üniversitesi Fen Bilimleri Enstitüsü Dergisi · doi:10.47495/okufbed.1100047 · Cited by 2 · see citing works
Мокросноп, В. М. (2018) · International Journal on Algae · doi:10.1615/interjalgae.v20.i2.40 · Cited by 2 · see citing works
Dong, Y., Quan, Y., Jiang, H., Hamouda, H. I., Lu, Z., Chen, Y., Li, C., & Mao, X. (2024) · Food Science and Human Wellness · doi:10.26599/fshw.2024.9250008 · Cited by 1 · see citing works
Takeda, T., Takahashi, M., Shimizu, M., Sugihara, Y., Saitoh, H., Fujisaki, K., Ishikawa, K., Utsushi, H., Kanzaki, E., Sakamoto, Y., et al. (2022) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2021.12.31.474618 · Cited by 1 · see citing works
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Yamamoto, K., KUWAYAMA, A., Tateishi, T., Suzuki, H., Suzuki, K., & SAKAMOTO, T. (2019) · Journal of the Japan Institute of Energy · doi:10.3775/jie.98.229
Abba, Z. (2019) · UTHM Institutional Repository (Universiti Tun Hussein Onn Malaysia)
Мокросноп, В. М. (2018) · Algologia · doi:10.15407/alg28.02.136
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Li, C., Chen, Z., Chen, Z., Lei, A., Liu, Q., & Wang, J. (2023) · CABI eBooks · doi:10.1079/9781800621954.0004
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Zhang, Y., Zhang, H. Y., Lou, T., Wang, S., Fan, Y., & Niu, S. (2026) · Critical Reviews in Food Science and Nutrition · doi:10.1080/10408398.2026.2628247
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- Authors
- Takeda, T., Nakano, Y., Takahashi, M., Konno, N., Sakamoto, Y., Arashida, R., Marukawa, Y., Yoshida, E., Ishikawa, T., & Suzuki, K.
- Affiliations
- Iwate Biotechnology Research Center (JP)
- Euglena Co., Ltd. (Japan) (JP)
- Shimane University (JP)
- Corresponding author
- Takeda, Takumi
- ORCID
- Venue
- Phytochemistry
- Publisher
- Elsevier BV
- Published
- 2015-08
- Volume
- 116
- Pages
- 21-27
- ISSN
- 0031-9422 (ISSN-L)
- OpenAlex
- W406434165
- Cited by
- 39
- References
- 33
- Citations by year
- 2026: 2 · 2025: 1 · 2024: 3 · 2023: 2 · 2022: 5 · 2021: 2 · 2020: 4 · 2019: 5 · 2018: 5 · 2017: 4 · 2016: 5 · 2015: 1
- FWCI
- 4.16
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Phytochemistry (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Iwate Prefecture
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biotechnology › Enzyme Production and Characterization
- Life Sciences › Agricultural and Biological Sciences › Plant Science › Polysaccharides and Plant Cell Walls
- Physical Sciences › Engineering › Biomedical Engineering › Biofuel production and bioconversion
- Keywords
- β-1,3-glucanase, Euglena gracilis, paramylon, laminarin, transglycosylation, enzyme purification, enzyme characterization, storage polysaccharides
- MeSH terms
- Electrophoresis, Polyacrylamide Gel, Glucan Endo-1,3-beta-D-Glucosidase, Euglena gracilis, Glycoside Hydrolases, Molecular Weight, beta-Glucans
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