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Characterization of an antifungal β-1,3-glucanase from Ficus microcarpa latex and comparison of plant and bacterial β-1,3-glucanases for fungal cell wall β-glucan degradation

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Abstract

Main conclusion

Each β-1,3-glucanase with antifungal activity or yeast lytic activity hydrolyzes different structures of β-1,3-glucans in the fungal cell wall, respectively.

Abstract

Plants express several glycoside hydrolases that target chitin and β-glucan in fungal cell walls and inhibit pathogenic fungal infection. An antifungal β-1,3-glucanase was purified from gazyumaru (Ficus microcarpa) latex, designated as GlxGluA, and the corresponding gene was cloned and expressed in Escherichia coli. The sequence shows that GlxGluA belongs to glycoside hydrolase family 17 (GH17). To investigate how GlxGluA acts to degrade fungal cell wall β-glucan, it was compared with β-1,3-glucanase with different substrate specificities. We obtained recombinant β-1,3-glucanase (designated as CcGluA), which belongs to GH64, from the bacterium Cellulosimicrobium cellulans. GlxGluA inhibited the growth of the filamentous fungus Trichoderma viride but was unable to lyse the yeast Saccharomyces cerevisiae. In contrast, CcGluA lysed yeast cells but had a negligible inhibitory effect on the growth of filamentous fungi. GlxGluA degraded the cell wall of T. viride better than CcGluA, whereas CcGluA degraded the cell wall of S. cerevisiae more efficiently than GlxGluA. These results suggest that the target substrates in fungal cell walls differ between GlxGluA (GH17 class I β-1,3-glucanase) and CcGluA (GH64 β-1,3-glucanase).

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Data availability

The sequence of the cDNA containing the GlxGluA gene has been deposited in the GenBank database under accession no. LC618823. The authors confirm that other experimental data are available and accessible via the main text and/or the supplemental data.

Abbreviations

CcGluA:

Cellulosimicrobium cellulans β-1,3-Glucanase

CM-curdlan:

Carboxymethyl curdlan

GlxGluA:

Gazyumaru latex glucanase-A

GH:

Glycoside hydrolase family

IFC:

Insoluble fraction of fungal cell

IPTG:

Isopropyl-β-D-1-thiogalactopyranoside

PDA:

Potato dextrose broth with agar

pNP:

para-Nitrophenol

References

  • Aimanianda V, Simenel C, Garnaud C, Clavaud C, Tada R, Barbin L, Mouyna I, Heddergott C, Popolo L, Ohya Y, Delepierre M, Latge JP (2017) The dual activity responsible for the elongation and branching of β-(1,3)-glucan in the fungal cell Wwall. Mbio 8:e00619-e717

    CAS  PubMed  PubMed Central  Google Scholar 

  • Beauvais A, Bruneau JM, Mol PC, Buitrago MJ, Legrand R, Latgé JP (2001) Glucan synthase complex of Aspergillus fumigatus. J Bacteriol 183:2273–2279

    CAS  PubMed  PubMed Central  Google Scholar 

  • Benítez T, Villa TG, Acha IG (1975) Chemical and structural differences in mycelial and regeneration walls of Trichoderma viride. Arch Microbiol 105:277–282

    PubMed  Google Scholar 

  • Branco AT, Dos Santos FB, Lourenco GF, Lopes Marques VC, Tavares Machado OL, Pereira MG, Aquino Almeida JC, de Souza Filho GA (2011) Induction of β-1,3-glucanase in seeds of maize defective-kernel mutant (827Kpro1). Protein Pept Lett 18:651–657

    CAS  PubMed  Google Scholar 

  • Cabib E (2009) Two novel techniques for determination of polysaccharide cross-links show that Crh1p and Crh2p attach chitin to both beta(1–6)- and beta(1–3)glucan in the Saccharomyces cerevisiae cell wall. Eukaryot Cell 8:1626–1636

    CAS  PubMed  PubMed Central  Google Scholar 

  • Cabib E, Blanco N, Grau C, Rodríguez-Peña JM, Arroyo J (2007) Crh1p and Crh2p are required for the cross-linking of chitin to beta(1–6)glucan in the Saccharomyces cerevisiae cell wall. Mol Microbiol 63:921–935

    CAS  PubMed  Google Scholar 

  • Cabib E, Blanco N, Arroyo J (2012) Presence of a large β(1–3)glucan linked to chitin at the Saccharomyces cerevisiae mother-bud neck suggests involvement in localized growth control. Eukaryot Cell 11:388–400

    CAS  PubMed  PubMed Central  Google Scholar 

  • Chye ML, Cheung KY (1995) β-1,3-Glucanase is highly-expressed in laticifers of Hevea brasiliensis. Plant Mol Biol 29:397–402

    CAS  PubMed  Google Scholar 

  • Dijkgraaf GJP, Abe M, Ohya Y, Bussey H (2002) Mutations in FskIp affect the cell wall content of β-1,3- and β-1,6-glucan in Saccharomyces cerevisiae. Yeast 19:671–690

    CAS  PubMed  Google Scholar 

  • Douglas CM, Foor F, Marrinan JA, Morin N, Nielsen JB, Dahl AM, Mazur P, Baginsky W, Li W, El-Sherbeini M, Clemas JA, Mandala SM, Frommer BR, Kurtz MB (1994) The Saccharomyces cerevisiae FKS1 (ETG1) gene encodes an integral membrane protein which is a subunit of 1,3-β-D-glucan synthase. Proc Natl Acad Sci USA 91:12907–12911

    CAS  PubMed  PubMed Central  Google Scholar 

  • Doxey AC, Yaish MW, Moffatt BA, Griffith M, McConkey BJ (2007) Functional divergence in the Arabidopsis beta-1,3-glucanase gene family inferred by phylogenetic reconstruction of expression states. Mol Biol Evol 24(4):1045–1055

    CAS  PubMed  Google Scholar 

  • Fang W, Sanz AB, Bartual SG, Wang B, Ferenbach AT, Farkaš V, Hurtado-Guerrero R, Arroyo J, van Aalten DMF (2019) Mechanisms of redundancy and specificity of the Aspergillus fumigatus Crh transglycosylases. Nat Commun 10:1–10

    Google Scholar 

  • Gastebois A, Mouyna I, Simenel C, Clavaud C, Coddeville B, Delepierre M, Latgé JP, Fontaine T (2010) Characterization of a new β(1–3)-glucan branching activity of Aspergillus fumigatus. J Biol Chem 285:2386–2396

    CAS  PubMed  Google Scholar 

  • Goldman RC, Sullivan PA, Zakula D, Capobianco JO (1995) Kinetics of β-1,3 glucan interaction at the donor and acceptor sites of the fungal glucosyltransferase encoded by the BGL2 gene. Eur J Biochem 227:372–378

    CAS  PubMed  Google Scholar 

  • Henrissat B, Davies G (1997) Structural and sequence-based classification of glycoside hydrolases. Curr Opin Struct Biol 7:637–644

    CAS  PubMed  Google Scholar 

  • Iseli B, Boller T, Neuhaus JM (1993) The N-terminal cysteine-rich domain of tobacco class I chitinase is essential for chitin binding but not for catalytic or antifungal activity. Plant Physiol 103:221–226

    CAS  PubMed  PubMed Central  Google Scholar 

  • Kalebina TS, Farkaš V, Laurinavichiute DK, Gorlovoy PM, Fominov GV, Bartek P, Kulaev IS (2003) Deletion of BGL2 results in an increased chitin level in the cell wall of Saccharomyces cerevisiae. Antonie Van Leeuwenhoek 84:179–184

    CAS  PubMed  Google Scholar 

  • Kaneko T, Kitamura K, Yamamoto Y (1969) Arthrobacter luteus nov. sp. isolated from brewery sewage. J Gen Appl Microbiol 15:317–326

    Google Scholar 

  • Kaneko T, Kitamura K, Yamamoto Y (1973) Susceptibilities of yeasts to yeast cell wall lytic enzyme of Arthrobacter luteus. Agric Biol Chem 37:2295–2302

    CAS  Google Scholar 

  • Kim Y-T, Kim E-H, Cheong C, Williams DL, Kim C-W, Lim S-T (2000) Structural characterization of β-d-(1→3, 1→6)-linked glucans using NMR spectroscopy. Carbohydr Res 328:331–341

    CAS  PubMed  Google Scholar 

  • Kitajima S, Aoki W, Shibata D, Nakajima D, Sakurai N, Yazaki K, Munakata R, Taira T, Kobayashi M, Aburaya S, Savadogo EH, Hibino S, Yano H (2018) Comparative multi-omics analysis reveals diverse latex-based defense strategies against pests among latex-producing organs of the fig tree (Ficus carica). Planta 247:1423–1438

    CAS  PubMed  Google Scholar 

  • Kitamura K (1982a) Re-examination of zymolyase purification. Agric Biol Chem 46:963–969

    CAS  Google Scholar 

  • Kitamura K (1982b) A protease that participates in yeast cell wall lysis during zymolyase digestion. Agric Biol Chem 46:2093–2099

    CAS  Google Scholar 

  • Kitamura K, Yamamoto Y (1972) Purification and properties of an enzyme, zymolyase, which lyses viable yeast cells. Arch Biochem Biophys 153:403–406

    CAS  PubMed  Google Scholar 

  • Kitamura K, Kaneko T, Yamamoto Y (1971) Lysis of viable yeast cells by enzymes of Arthrobacter luteus. Arch Biochem Biophys 145:402–404

    CAS  PubMed  Google Scholar 

  • Kollár R, Petráková E, Ashwell G, Robbins PW, Cabib E (1995) Architecture of the yeast cell wall. The linkage between chitin and beta(1–>3)-glucan. J Biol Chem 270(3):1170–1178

    PubMed  Google Scholar 

  • Konno K (2011) Plant latex and other exudates as plant defense systems: Roles of various defense chemicals and proteins contained therein. Phytochemistry 72:1510–1530

    CAS  PubMed  Google Scholar 

  • Konno K, Hirayama C, Nakamura M, Tateishi K, Tamura Y, Hattori M, Kohno K (2004) Papain protects papaya trees from herbivorous insects: role of cysteine proteases in latex. Plant J 37:370–378

    CAS  PubMed  Google Scholar 

  • Kumagai Y, Okuyama M, Kimura A (2016) Heat treatment of curdlan enhances the enzymatic production of biologically active β-(1,3)-glucan oligosaccharides. Carbohydr Polym 146:396–401

    CAS  PubMed  Google Scholar 

  • Latgé JP, Beauvais A (2014) Functional duality of the cell wall. Curr Opin Microbiol 20:111–117

    PubMed  Google Scholar 

  • Leah R, Tommerup H, Svendsen I, Mundy J (1991) Biochemical and molecular characterization of three barley seed proteins with antifungal properties. J Biol Chem 266:1564–1573

    CAS  PubMed  Google Scholar 

  • Lesage G, Bussey H (2006) Cell wall assembly in Saccharomyces cerevisiae. Microbiol Mol Biol 70:317–343. https://doi.org/10.1128/MMBR.00038-05

    Article  CAS  Google Scholar 

  • Leubner-Metzger G, Meins F Jr (2000) Sense transformation reveals a novel role for class I beta-1, 3-glucanase in tobacco seed germination. Plant J 23:215–221

    CAS  PubMed  Google Scholar 

  • Lipke PN, Ovalle R (1998) Cell wall architecture in yeast: New structure and new challenges. J Bacteriol 180:3735–3740

    CAS  PubMed  PubMed Central  Google Scholar 

  • Liu B, Lu Y, Xin Z, Zhang Z (2009) Identification and antifungal assay of a wheat β-1,3-glucanase. Biotechnol Lett 31:1005–1010

    CAS  PubMed  Google Scholar 

  • Mauch F, Mauch-Mani B, Boller T (1988) Antifungal hydrolases in pea tissue. Plant Physiol 88:936–942

    CAS  PubMed  PubMed Central  Google Scholar 

  • McIntosh M, Stone BA, Stanisich VA (2005) Curdlan and other bacterial (1→3)-β-D-glucans. Appl Microbiol Biotechnol 68:163–173

    CAS  PubMed  Google Scholar 

  • Moir GFJ (1959) Ultracentrifugation and staining of Hevea latex. Nature 184:1626–1628

    Google Scholar 

  • Mondal KK, Bhattacharya RC, Koundal KR, Chatterjee SC (2007) Transgenic Indian mustard (Brassica juncea) expressing tomato glucanase leads to arrested growth of Alternaria brassicae. Plant Cell Rep 26:247–252

    CAS  PubMed  Google Scholar 

  • Moravčíková J, Margetínyová D, Gálová Z, Zur I, Gregorová Z, Zimová M, Boszorádová E, Matušíková I (2016) Beta-1,3-glucanase activities in wheat and relative species. Nov Biotechnol Chim 15:122–132

    Google Scholar 

  • Pardini G, De Groot PW, Coste AT, Karababa M, Klis FM, de Koster CG, Sanglard D (2006) The CRH family coding for cell wall glycosylphosphatidylinositol proteins with a predicted transglycosidase domain affects cell wall organization and virulence of Candida albicans. J Biol Chem 281:40399–40411

    CAS  PubMed  Google Scholar 

  • Schlumbaum A, Mauch F, Vögeli U, Boller T (1986) Plant chitinases are potent inhibitors of fungal growth. Nature 324:365–367

    CAS  Google Scholar 

  • Sela-Buurlage MB, Ponstein AS, Bres-Vloemans SA, Melchers LS, Van Den Elzen PJM, Cornelissen BJC (1993) Only specific tobacco (Nicotiana tabacum) chitinases and β-1,3-glucanases exhibit antifungal activity. Plant Physiol 101:857–863

    CAS  PubMed  PubMed Central  Google Scholar 

  • Shen SH, Chretien P, Bastien L, Slilaty SN (1991) Primary sequence of the glucanase gene from Oerskovia xanthineolytica. Expression and purification of the enzyme from Escherichia coli. J Biol Chem 266:1058–1063

    CAS  PubMed  Google Scholar 

  • Shinshi H, Wenzler H, Neuhaus J-M, Felix G, Hofsteenge J, Meins F (1988) Evidence for N- and C-terminal processing of a plant defense-related enzyme: Primary structure of tobacco prepro- -1,3-glucanase. Proc Natl Acad Sci USA 85:5541–5545

    CAS  PubMed  PubMed Central  Google Scholar 

  • Sridevi G, Parameswari C, Sabapathi N, Raghupathy V, Veluthambi K (2008) Combined expression of chitinase and β-1,3-glucanase genes in indica rice (Oryza sativa L.) enhances resistance against Rhizoctonia solani. Plant Sci 175:283–290

    CAS  Google Scholar 

  • Suarez V, Staehelin C, Arango R, Holtorf H, Hofsteenge J, Meins F (2001) Substrate specificity and antifungal activity of recombinant tobacco class I chitinases. Plant Mol Biol 45:609–618

    CAS  PubMed  Google Scholar 

  • Taira T (2010) Structures and antifungal activity of plant chitinases. J Appl Glycosci 57:167–176

    CAS  Google Scholar 

  • Taira T, Ohnuma T, Yamagami T, Aso Y, Ishiguro M, Ishihara M (2002) Antifungal activity of rye (Secale cereale) seed chitinases: the different binding manner of class I and class II chitinases to the fungal cell walls. Biosci Biotechnol Biochem 66:970–977

    CAS  PubMed  Google Scholar 

  • Taira T, Ohdomari A, Nakama N, Shimoji M, Ishihara M (2005) Characterization and antifungal activity of gazyumaru (Ficus microcarpa) latex chitinases: both the chitin-binding and the antifungal activities of class I chitinase are reinforced with increasing ionic strength. Biosci Biotechnol Biochem 69(4):811–818. https://doi.org/10.1271/bbb.69.811

    Article  CAS  PubMed  Google Scholar 

  • Takashima T, Henna H, Kozome D, Kitajima S, Uechi K, Taira T (2021) cDNA cloning, expression, and antifungal activity of chitinase from Ficus microcarpa latex: difference in antifungal action of chitinase with and without chitin-binding domain. Planta 253:120. https://doi.org/10.1007/s00425-021-03639-8

    Article  CAS  PubMed  Google Scholar 

  • van Kan JAL, Joosten MHAJ, Wagemakers CAM, van den Berg-Velthuis GCM, de Wit PJGM (1992) Differential accumulation of mRNAs encoding extracellular and intracellular PR proteins in tomato induced by virulent and avirulent races of Cladosporium fulvum. Plant Mol Biol 20:513–527

    PubMed  Google Scholar 

  • Wan L, Zha W, Cheng X, Liu C, Lv L, Liu C, Wang Z, Du B, Chen R, Zhu L, He G (2011) A rice β-1,3-glucanase gene Osg1 is required for callose degradation in pollen development. Planta 233:309–323

    CAS  PubMed  Google Scholar 

  • Wojtkowiak A, Witek K, Hennig J, Jaskolski M (2013) Structures of an active-site mutant of a plant 1,3-β-glucanase in complex with oligosaccharide products of hydrolysis. Acta Crystallogr D Biol Crystallogr 69:52–62

    CAS  PubMed  Google Scholar 

  • Wu HM, Liu SW, Hsu MT, Hung CL, Lai CC, Cheng WC, Wang HJ, Li YK, Wang WC (2009) Structure, mechanistic action, and essential residues of a GH-64 enzyme, laminaripentaose-producing β-1,3-glucanase. J Biol Chem 284:26708–26715

    CAS  PubMed  PubMed Central  Google Scholar 

  • Yun DJ, Zhao Y, Pardo JM, Narasimhan ML, Damsz B, Lee H, Abad LR, D’Urzo MP, Hasegawa PM, Bressan RA (1997) Stress proteins on the yeast cell surface determine resistance to osmotin, a plant antifungal protein. Proc Natl Acad Sci USA 94:7082–7087

    CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

This work was supported by JSPS KAKENHI Grant Number JP23H00247.

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Authors and Affiliations

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

    Tomoya Takashima, Nao Komori, Keiko Uechi & Toki Taira

  2. Graduate School of Agricultural Science, Kagoshima University, Kagoshima, 890-8580, Japan

    Toki Taira

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  1. Tomoya Takashima
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Contributions

TTak and NK: performed the experiments. TTak and TTai: wrote the manuscript. TTak, TTai and KU: designed the experiments and thoroughly revised the manuscript. TTai: coordinated the research project. All the authors read and approved the manuscript.

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Correspondence to Toki Taira.

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Takashima, T., Komori, N., Uechi, K. et al. Characterization of an antifungal β-1,3-glucanase from Ficus microcarpa latex and comparison of plant and bacterial β-1,3-glucanases for fungal cell wall β-glucan degradation. Planta 258, 116 (2023). https://doi.org/10.1007/s00425-023-04271-4

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