Purification and characterization of an extracellular (1 → 6)-<i>β</i>-glucanase from the filamentous fungus <i>Acremonium persicinum</i>

Pitson, Stuart M.; Seviour, Robert J.; McDougall, Barbara M.; Stone, Bruce; Sadek, Maruse

doi:10.1042/bj3160841

Cited by 38 publications

(15 citation statements)

References 35 publications

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“…␤-1,6-Glucanases have been purified from a number of fungal species including Acremonium persicinum, Neurospora crassa, Rhizopus chinensis, Penicillium spp., and Saccharomycopsis fibuligera (Yamamoto et al, 1974;Santos et al, 1979;Schep et al, 1984;Hiura et al, 1987;Mulenga and Berry, 1994;Pitson et al, 1996). The only DNA sequences reported to date, however, are from two mycoparasitic fungi, T. harzianum and T. virens (Lora et al, 1995;Kim et al, 2002).…”

Section: Discussionmentioning

confidence: 99%

Endophytic Fungal β-1,6-Glucanase Expression in the Infected Host Grass

Moy

Sullivan

et al. 2002

Plant Physiology

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Mutualistic fungal endophytes infect many grass species and often confer benefits to the hosts such as reduced herbivory by insects and animals. The physiological interactions between the endophytes and their hosts have not been well characterized. Fungal-secreted proteins are likely to be important components of the interaction. In the interaction between Poa ampla and the endophyte Neotyphodium sp., a fungal ␤-1,6-glucanase is secreted into the apoplast, and activity of the enzyme is detectable in endophyte-infected plants. Sequence analysis indicates the ␤-1,6-glucanase is homologous to enzymes secreted by the mycoparasitic fungi Trichoderma harzianum and Trichoderma virens. DNA gel-blot analysis indicated the ␤-1,6-glucanase was encoded by a single gene. As a secreted protein, the ␤-1,6-glucanase may have a nutritional role for the fungus. In culture, ␤-1,6-glucanase activity was induced in the presence of ␤-1,6-glucans. From RNA gel blots, similar ␤-1,6-glucanases were expressed in tall fescue (Festuca arundinacea Schreb.) and Chewings fescue (Festuca rubra L. subsp. fallax [Thuill] Nyman) infected with the endophyte species Neotyphodium coenophialum and Epichloë festucae, respectively.Fungal endophytes of the genus Neotyphodium (formerly Acremonium; Glenn et al., 1996) infect many grass species, some of which are important turf and forage grasses. The fungi colonize the intercellular spaces of the aerial plant parts but do not invade the plant cells. The endophyte-grass associations are generally considered to be mutualistic symbioses (Clay, 1988). In many associations, the production of alkaloids by the fungus results in reduced herbivory by insects and animals, thus benefiting the host (Breen, 1994;Bush et al., 1997). The fungi benefit from the access to nutrients provided by the plants.Within the past 20 years, considerable knowledge has been gained on the synthesis and effects of alkaloids, the genetics and taxonomic relationships of endophytes, and the ecological effects of endophyte infection (Clay, 1990;Siegel and Schardl, 1991;Schardl, 1996;Bush et al., 1997). The physiological aspects of the endophyte-grass interactions have not, however, been well characterized in any system. We are investigating the physiology of the fungus-grass interaction with the long-range objective of eventually being able to manipulate agriculturally important interactions. We are using the Poa ampla cv Service (big bluegrass)/Neotyphodium sp. interaction as a model system for the grass/fungus interaction (Lindstrom et al., 1993). P. ampla is apomictic, so we have a ready supply of plants of identical genotype. We also have uninfected plants of the identical genotype, which were identified in older seed lots in which the endophyte had lost viability.Almost nothing is known of the proteins relevant to the interaction between the plant hosts and the fungal endophytes. We are interested in fungalsecreted proteins because they are likely to be important components of the mutualistic interaction because they are located at the...

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Section: Discussionmentioning

confidence: 99%

Endophytic Fungal β-1,6-Glucanase Expression in the Infected Host Grass

Moy

Sullivan

et al. 2002

Plant Physiology

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“…For determination of Michaelis-Menten constants on laminarin and pustulan, the initial velocities were measured for substrate concentrations ranging from 1.25 to 20 mg ml Ϫ1 . The degrees of polymerization of pustulan and laminarin used to calculate their molar concentrations were 94 (22) and 28 (17), respectively. The kinetic parameters were estimated using weighted nonlinear least-squares regression analysis with Grafit software (Erithacus Software, Horley, United Kingdom).…”

Section: Methodsmentioning

confidence: 99%

Characterization of a Broad-Specificity β-Glucanase Acting on β-(1,3)-, β-(1,4)-, and β-(1,6)-Glucans That Defines a New Glycoside Hydrolase Family

Lafond

Navarro

Haon

et al. 2012

Appl Environ Microbiol

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Here we report the cloning of the Pa_3_10940 gene from the coprophilic fungus Podospora anserina, which encodes a C-terminal family 1 carbohydrate binding module (CBM1) linked to a domain of unknown function. The function of the gene was investigated by expression of the full-length protein and a truncated derivative without the CBM1 domain in the yeast Pichia pastoris. Using a library of polysaccharides of different origins, we demonstrated that the full-length enzyme displays activity toward a broad range of ␤-glucan polysaccharides, including laminarin, curdlan, pachyman, lichenan, pustulan, and cellulosic derivatives. Analysis of the products released from polysaccharides revealed that this ␤-glucanase is an exo-acting enzyme on ␤-(1,3)-and ␤-(1,6)-linked glucan substrates and an endo-acting enzyme on ␤-(1,4)-linked glucan substrates. Hydrolysis of short ␤-(1,3), ␤-(1,4), and ␤-(1,3)/␤-(1,4) gluco-oligosaccharides confirmed this striking feature and revealed that the enzyme performs in an exo-type mode on the nonreducing end of gluco-oligosaccharides. Excision of the CBM1 domain resulted in an inactive enzyme on all substrates tested. To our knowledge, this is the first report of an enzyme that displays bifunctional exo-␤-(1,3)/(1,6) and endo-␤-(1,4) activities toward beta-glucans and therefore cannot readily be assigned to existing Enzyme Commission groups. The amino acid sequence has high sequence identity to hypothetical proteins within the fungal taxa and thus defines a new family of glycoside hydrolases, the GH131 family.

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“…Therefore, since the rate o" hydrolysis must be greater than that of mutarotation, these substrates are not particularly suitable for stereochemical aaalysis of endo-arabinanases where very rapid mutarotation c" the hydrolysis products (arabinofuranosyl oligosaccharides) as observed to occur. Since several recent reports have desc ibed the successful use of natural polymers as substrates for slereochemical analysis with 1H-NMR [37][38][39], we have empoyed linear (apple) (1 ~ 5)-Ct-L-arabinan for this purpose in ti e current study. Fig.…”

Section: Endo-arabinanasesmentioning

confidence: 99%

Stereochemical course of hydrolysis catalyzed by arabinofuranosyl hydrolases

Pitson¹,

Voragen²,

Beldman³

1996

FEBS Letters

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1~ IntroductionEnzymic hydrolysis of glycosidic linkages occurs via two m ~jor mechanisms, which result in either net retention or inversion of anomeric configuration [1]. Both hydrolytic mechmisms involve general acid catalysis and require two critical reddues (a proton donor and a nucleophile/base), which in m 9st glycosyl hydrolases are aspartate and/or glutamate resid~es 2.1.99)hydrolyse the (1 ~ 5)-C~-L-arabinofuranosyl linkages in linear arabinan in a random pattern, initially releasing arabinotriose and larger arabino-oligosaccharides as the major hydrolysis products [22]. In contrast, C~-L-arabinofuranosidases (a-L-arabinofuranoside arabinofuranohydrolase, EC 3.2.1.55) hydrolyse nonreducing C~-L-arabinofuranosyl residues from arabinofuranosecontaining substrates, releasing arabinose as the only hydrolysis product [21]. Two major types of C~-L-arabinofuranosidases can be classified according to substrate specificity, and are typified by the two enzymes isolated from Aspergillus niger [21,24]. Arabinofuranosidase A is only active against small substrates, like 4-nitrophenyl a-L-arabinofuranoside (pNPA) and short-chain arabino-oligosaccharides, while arabinofuranosidase B has similar activity on these substrates, but is also able to hydrolyse polymeric substrates like branched arabinan and arabinoxylan. Recently, other enzymes have been isolated [25,26] that are specific for arabinofuranosyl residues in arabinoxylan, and are termed arabinoxylan arabinohydrolases ((1 ~4)-[3-D-arabinoxylan arabinofuranohydrolase, EC 3.2.1.) (AXH). The AXH from Aspergillus awamori has been studied in some detail [23,25,27] and found to only cleave arabinofuranose from single-substituted xylopyranosyl residues in the xylan backbone, and has only low activity against substrates like pNPA or branched arabinan. Similar enzymes from B/fidobacterium adolescentis [26] and Trichoderma reesei (R. Kavitha, H. Gruppen and A.G.J. Voragen, unpublished) have also been recently isolated that appear to have a slightly different specificity since they are active against arabinofuranosyl groups linked to double-substituted xylopyranosyl residues, and are therefore termed AXH-d.Apart from studies with the c~-e-arabinofuranosidases from Monilinia fructigena [28,29], there has been little work examining the mechanisms by which these enzymes cleave arabinofuranosyl linkages. In this current study the stereochemical course of hydrolysis catalyzed by endo-(l ~5)-Ot-L-arabina-

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Purification and characterization of an extracellular (1 → 6)-β-glucanase from the filamentous fungus Acremonium persicinum

Cited by 38 publications

References 35 publications

Endophytic Fungal β-1,6-Glucanase Expression in the Infected Host Grass

Endophytic Fungal β-1,6-Glucanase Expression in the Infected Host Grass

Characterization of a Broad-Specificity β-Glucanase Acting on β-(1,3)-, β-(1,4)-, and β-(1,6)-Glucans That Defines a New Glycoside Hydrolase Family

Stereochemical course of hydrolysis catalyzed by arabinofuranosyl hydrolases

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