Application of carbohydrate arrays coupled with mass spectrometry to detect activity of plant-polysaccharide degradative enzymes from the fungus Aspergillus niger

The aim of this study was to develop and characterize antigens for the diagnosis of aspergillosis. Nine strains of Aspergillus species Aspergillus fumigatus , Aspergillus flavus , and Aspergillus niger were grown in Sabouraud and Smith broth to produce exoantigens. The antigens were tested by immunodiffusion against sera from patients with aspergillosis and other systemic mycoses. The protein fraction of the antigens was detected by SDS–PAGE; Western blot and representative bands were assessed by mass spectrometry coupled to a nano Acquity UltraPerformance LC and analyzed by the Mascot search engine. Concurrently, all sera were tested with Platelia Aspergillus EIA. The most reactive antigens to sera from patients infected by A. fumigatus were produced by A. fumigatus MG2 Sabouraud and pooled A. fumigatus Sabouraud samples, both with a sensitivity of 93% and specificity of 100% and 97%, respectively. Aspergillus niger and A. flavus antigens were reactive against A. niger and A. flavus sera, each one with a sensitivity and specificity of 100%. Two proteins, probably responsible for antigenic activity, β-glucosidase in A. fumigatus and α-amylase in A. niger were attained. The commercial kit had a specificity of 22%, sensitivity of 100%, positive predictive value of 48%, and negative predictive value of 100%. The antigens produced showed high sensitivity and specificity and can be exploited for diagnostics of aspergilloma.

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Shotgun Proteomics of Aspergillus niger Microsomes upon d-Xylose Induction

Applied and Environmental Microbiology ◽

10.1128/aem.00482-10 ◽

2010 ◽

Vol 76 (13) ◽

pp. 4421-4429 ◽

Cited By ~ 31

Author(s):

José Miguel P. Ferreira de Oliveira ◽

Mark W. J. van Passel ◽

Peter J. Schaap ◽

Leo H. de Graaff

Keyword(s):

Mass Spectrometry ◽

Aspergillus Niger ◽

Protein Secretion ◽

Secretory Pathway ◽

Extracellular Enzymes ◽

Protein Composition ◽

Shotgun Proteomics ◽

Small Gtpases ◽

Hypothetical Proteins ◽

Extracellular Environment

ABSTRACT Protein secretion plays an eminent role in cell maintenance and adaptation to the extracellular environment of microorganisms. Although protein secretion is an extremely efficient process in filamentous fungi, the mechanisms underlying protein secretion have remained largely uncharacterized in these organisms. In this study, we analyzed the effects of the d-xylose induction of cellulase and hemicellulase enzyme secretion on the protein composition of secretory organelles in Aspergillus niger. We aimed to systematically identify the components involved in the secretion of these enzymes via mass spectrometry of enriched subcellular microsomal fractions. Under each condition, fractions enriched for secretory organelles were processed for tandem mass spectrometry, resulting in the identification of peptides that originate from 1,081 proteins, 254 of which—many of them hypothetical proteins—were predicted to play direct roles in the secretory pathway. d-Xylose induction led to an increase in specific small GTPases known to be associated with polarized growth, exocytosis, and endocytosis. Moreover, the endoplasmic-reticulum-associated degradation (ERAD) components Cdc48 and all 14 of the 20S proteasomal subunits were recruited to the secretory organelles. In conclusion, induction of extracellular enzymes results in specific changes in the secretory subproteome of A. niger, and the most prominent change found in this study was the recruitment of the 20S proteasomal subunits to the secretory organelles.

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Gas chromatography-mass spectrometry analysis of principal lipid-soluble components of Pinellia ternate fermented with Bacillus subtilis, Aspergillus niger and Meyerozyma guillermondii

Tropical Journal of Pharmaceutical Research ◽

10.4314/tjpr.v18i6.23 ◽

2021 ◽

Vol 18 (6) ◽

pp. 1305-1303

Author(s):

Lin Wang ◽

Jiabin Sun ◽

Meiying Yue ◽

Jin Wang ◽

Qinwan Huang ◽

...

Keyword(s):

Mass Spectrometry ◽

Gas Chromatography ◽

Bacillus Subtilis ◽

Aspergillus Niger ◽

Gas Chromatography Mass Spectrometry ◽

Chemical Components ◽

Fermentation Products ◽

Chromatography Mass Spectrometry ◽

Pure Cultures ◽

Soluble Components

Purpose: To study the differences in lipid-soluble compounds from naturally-fermented Rhizoma Pinelliae fermentata (BXQ) samples, and fermentation products of BXQ using pure cultures of Bacillus subtilis, Aspergillus niger, and Meyerozyma guillermondii. Methods: First, unfermented BXQ (CTFJ-Q), traditional, naturally-fermented BXQ (CTFJ-H), and fermentation products of BXQ using pure cultures of Bacillus subtilis (XJFJ), Aspergillus niger (MJFJ), and Meyerozyma guillermondii (JMJFJ) were obtained. Their lipid-soluble components were then analyzed using gas chromatography-mass spectrometry (GC-MS) technology and principal component analysis (PCA). Results: GC-MS results showed that there were 26, 24, 27, 31 and 32 types of chemical components in CTFJ-Q, CTFJ-H, XJFJ, MJFJ and JMJFJ, respectively. Furthermore, PCA revealed that samples obtained using fermentation with pure cultures of the three microorganisms had unique chemical components. Conclusion: These results suggest that the microorganisms used for fermentation greatly influence the lipid-soluble components of BXQ. This finding is considered beneficial for the optimization of BXQ fermentation process.

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Comprehensive Study on the chemical profile and anti-tumor activity of secondary metabolites produced by Aspergillus niger

Current Pharmaceutical Analysis ◽

10.2174/1573412917666210121152317 ◽

2021 ◽

Vol 17 ◽

Author(s):

Yung-Husan Chen ◽

Pinghong Chen ◽

Ching-Hsu Yang ◽

Xiuna Wu ◽

Lianzhong Luo ◽

...

Keyword(s):

Mass Spectrometry ◽

Aspergillus Niger ◽

Inhibitory Effect ◽

Specific Tumor ◽

Orbitrap Mass Spectrometry ◽

Hela S3 ◽

Diagnostic Ions ◽

First Time ◽

Anti Tumor Activity

Objective: Chemical investigation of the extract of sponge-derived fungus, Aspergillus niger, was performed by liquid chromatography coupled with QExactive mass spectrometry for the first time. Method: A total number of 444 constituents were detected, 288 of which were identiﬁed positively or tentatively by the comprehensive utilization of accurate molecular weight and fragmentation information acquired from quadrupole orbitrap mass spectrometry. The identified compounds were divided into several types, namely, organic acid, alkaloid, saccharide, amino acid and cyclopeptide, terpenoid, polyketone, phenylpropanoid and other types of compounds. Systematic diagnostic ions and featured fragment patterns were summarized for each type, based on which 8 novel compounds belonging to the same type were characterized. Results: This work provided a rapid approach for the research of microconstituents in a complex analyte. Furthermore, the anti-tumor activity of the extract was evaluated on two different cell lines—Bel-7402 and Hela-S3 in vitro. The tumor-inhibitory effect of the Aspergillus niger extract was confirmed, and may be mainly derived from its pro-apoptotic action. Moreover, the extract exerted more significant cytotoxicity in Bel-7402 cells than Hela-S3 cells, indicative of its selectivity on specific tumor cells. Conclusion: The evidence suggested that the Aspergillus niger extract may potentially serve as a remedy for prevention and therapy of hepatic and breast carcinoma.

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