An extracellular β-galactofuranosidase from Aspergillus niger and its use as a tool for glycoconjugate analysis

Wallis, Gregg; Hemming, Frank W.; Peberdy, John F.

doi:10.1016/s0304-4165(00)00150-1

Cited by 30 publications

(36 citation statements)

References 31 publications

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“…A comparison between enzyme levels shows that A. fumigatus (this study) produced about the same level as the Penicillium species (5 to 50 U/liter) but not as much as A. niger (250 U/liter) (56). However, growth conditions and substrates differed between different studies.…”

Section: Vol 44 2006 Release Of Surrogate Markers By Aspergillus Fumentioning

confidence: 57%

“…The production of this enzyme seemed to be medium dependent, with glucose as a repressor (10). Furthermore, a galactofuranosidase was purified from A. niger with an optimum pH of 3 to 4 (56). The results of this study suggest that ␤-galactofuranosidase activity is involved in the decrease of galf epitopes in the culture supernatant.…”

Section: Vol 44 2006 Release Of Surrogate Markers By Aspergillus Fumentioning

confidence: 83%

“…A specific ␤-D-galactofuranosidase has been detected in only a few species of fungi: P. fellutanum (44), Penicillum and Aspergillus species (10), T. harzianum (53), Helminthosporium sacchari (11), and A. niger (56). A comparison between enzyme levels shows that A. fumigatus (this study) produced about the same level as the Penicillium species (5 to 50 U/liter) but not as much as A. niger (250 U/liter) (56).…”

Section: Vol 44 2006 Release Of Surrogate Markers By Aspergillus Fumentioning

confidence: 99%

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In Vitro Release by Aspergillus fumigatus of Galactofuranose Antigens, 1,3-β- d -Glucan, and DNA, Surrogate Markers Used for Diagnosis of Invasive Aspergillosis

et al. 2006

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Aspergillus markers are becoming increasingly important for the early diagnosis of invasive aspergillosis. The kinetics of release of these surrogate markers, however, is largely unknown. We investigated the release of ␤-(1-5)-galactofuranosyl (galf) antigens (Platelia Aspergillus), 1,3-␤-D-glucan (BG) (Fungitell), and DNA (PCR) in an in vitro model of Aspergillus fumigatus. The results showed that release is correlated to the growth phase of the fungus, which depends on available nutrients. Whereas galf antigens and BG are released during logarithmic growth, DNA is released only after mycelium breakdown. During early logarithmic growth, galf antigens seem to be released somewhat earlier than BG. Furthermore, galf antigen concentrations of more than 120,000 times the serum cutoff value (0.5 ng/ml) can be measured, while BG concentrations reach a value only 978 times the serum cutoff value (60 pg/ml). During lytical growth, release of galf antigens further increased to a maximum level, which depended on pH. After that, the concentration of galf antigens stayed high (pH 7.4) or decreased to zero within 4 days (pH 5.0). In contrast to galf antigens, BG concentration decreased after 1 day of growth. The decrease of galf components seems to be due to the enzyme ␤-galactofuranosidase, which is able to destroy galf epitopes and whose activity fluctuates in the culture filtrates in parallel with galf antigen concentration. Fungal DNA seems to be released only due to autolysis caused by nutrient limitation. In conclusion, several factors clearly influence the release of surrogate markers in vitro. These same factors might also play a role at the infection site of Aspergillus disease in humans.Invasive aspergillosis (IA) has become a leading cause of death among immunocompromised hosts, including transplant patients, those treated for hematological malignancy, and those treated with high-dose corticosteroids (41, 49). In addition, IA is increasingly observed in the nonneutropenic phase after hematopoietic stem cell transplantation and in nonclassic settings, such as intensive care units with critically ill patients (13, 16). The high mortality is due partly to the difficulty in establishing a diagnosis at an early stage of infection, since presenting symptoms are nonspecific and sensitivity of cultures is low. Techniques to improve timely diagnosis have focused on the detection of circulating surrogate markers released by the fungus (43,45,50). With the development of nonculture-based methods, such as PCR and antigen detection, circulating markers can be detected at an early stage of infection in patients with invasive disease (12, 27, 28).The commercially available sandwich enzyme-linked immunosorbent assay (ELISA) (Platelia Aspergillus [PA]; Bio-Rad, Marnes-la-Coquette, France) is based on the rat immunoglobulin M monoclonal antibody EB-A2, which binds the ␤-(1-5)-galactofuranosyl (galf) side chains of the Aspergillus galactomannan (GM) molecule (24,50,51). In addition to GM, fungal glycoproteins also react with the EB-...

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Section: Vol 44 2006 Release Of Surrogate Markers By Aspergillus Fumentioning

confidence: 57%

Section: Vol 44 2006 Release Of Surrogate Markers By Aspergillus Fumentioning

confidence: 83%

Section: Vol 44 2006 Release Of Surrogate Markers By Aspergillus Fumentioning

confidence: 99%

See 1 more Smart Citation

In Vitro Release by Aspergillus fumigatus of Galactofuranose Antigens, 1,3-β- d -Glucan, and DNA, Surrogate Markers Used for Diagnosis of Invasive Aspergillosis

et al. 2006

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“…This issue has not been investigated yet. Molecules recognized (32,36,37). (ii) When present, Galf is at the nonreducing terminal position.…”

Section: Discussionmentioning

confidence: 99%

Galactomannoproteins of Aspergillus fumigatus

et al. 2005

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Galactofuranose-containing molecules have been repeatedly shown to be important antigens among human fungal pathogens, including Aspergillus fumigatus. Immunogenic galactofuran determinants have been poorly characterized chemically, however. We reported here the characterization of two glycoproteins of A. fumigatus with an N-glycan containing galactofuranose. These proteins are a phospholipase C and a phytase. Chemical characterization of the N-glycan indicates that it is a mixture of Hex 5-13 HexNAc 2 oligosaccharides, the major molecular species corresponding to Hex 6-8 HexNAc 2 . The N-glycan contained one galactofuranose unit that was in a terminal nonreducing position attached to the 2 position of Man. This single terminal nonreducing galactofuranose is essential for the immunoreactivity of the N-glycans assessed either with a monoclonal antibody that recognizes a tetra-␤-1,5-galactofuran chain of galactomannan or with Aspergillus-infected patient sera.

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“…An example of such an unattributed activity is the enzymatic degradation of Gal f -containing glycoconjugates. This activity has been described for Aspergillus niger (Cousin et al , 1989 ;Wallis et al , 2001 ), for several other fi lamentous fungi (Daley and Strobel , 1983 ;Cousin et al , 1989 ;Reyes et al , 1992 ;Van Bruggen -Van Der Lugt et al, 1992 ), and for a prokaryote, Bacillus sp. (Ramli et al , 1995 ).…”

Section: Introductionmentioning

confidence: 77%

Fungal α-arabinofuranosidases of glycosyl hydrolase families 51 and 54 show a dual arabinofuranosyl- and galactofuranosyl-hydrolyzing activity

Tefsen¹,

Lagendijk²,

Park

et al. 2012

Biological Chemistry

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Aspergillus niger possesses a galactofuranosidase activity, however, the corresponding enzyme or gene encoding this enzyme has never been identifi ed. As evidence is mounting that enzymes exist with affi nity for both arabinofuranose and galactofuranose, we investigated the possibility that α -l -arabinofuranosidases, encoded by the abfA and abfB genes, are responsible for the galactofuranosidase activity of A. niger . Characterization of the recombinant AbfA and AbfB proteins revealed that both enzymes do not only hydrolyze p -nitrophenyl-α -l -arabinofuranoside (pNp-α -Ara f ) but are also capable of hydrolyzing p -nitrophenyl-β -d -galactofuranoside (pNp-β -Gal f ). Molecular modeling of the AbfB protein with pNp-β -Gal f confi rmed the possibility for AbfB to interact with this substrate, similarly as with pNp-α -Ara f . We also show that galactomannan, a cell wall compound of A. niger , containing β -linked terminal and internal galactofuranosyl moieties, can be degraded by an enzyme activity that is present in the supernatant of inulin-grown A. niger . Interestingly, purifi ed AbfA and AbfB did not show this hydrolyzing activity toward A. niger galactomannan. In summary, our studies demonstrate that AbfA and AbfB, α -l -arabinofuranosidases from different families, both contain a galactofuranose (Galf) -hydrolyzing activity. In addition, our data support the presence of a Gal fhydrolase activity expressed by A. niger that is capable of degrading fungal galactomannan.

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An extracellular β-galactofuranosidase from Aspergillus niger and its use as a tool for glycoconjugate analysis

Cited by 30 publications

References 31 publications

In Vitro Release by Aspergillus fumigatus of Galactofuranose Antigens, 1,3-β- d -Glucan, and DNA, Surrogate Markers Used for Diagnosis of Invasive Aspergillosis

In Vitro Release by Aspergillus fumigatus of Galactofuranose Antigens, 1,3-β- d -Glucan, and DNA, Surrogate Markers Used for Diagnosis of Invasive Aspergillosis

Galactomannoproteins of Aspergillus fumigatus

Fungal α-arabinofuranosidases of glycosyl hydrolase families 51 and 54 show a dual arabinofuranosyl- and galactofuranosyl-hydrolyzing activity

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