Abstract
Several species of fungus from the genus Aspergillus are implicated in pulmonary infections in immunocompromised patients. Broad screening methods for fungal infections are desirable, as cultures require a considerable amount of time to provide results. Herein, we developed degradation and detection methods to produce and detect D-glucosamine (GlcN) from Aspergillus niger, a species of filamentous fungus. Ultimately, these techniques hold the potential to contribute to the diagnosis of pulmonary fungal infections in immunocompromised patients. In the following studies, we produced GlcN from fungal-derived chitin to serve as a marker for Aspergillus niger. To accomplish this, A. niger cells were lysed and subjected to a hydrochloric acid degradation protocol. Products were isolated, reconstituted in aqueous solutions, and analyzed using hydrophilic interaction liquid chromatography (HILIC) in tandem with electrospray ionization time-of-flight mass spectrometry. Our results indicated that GlcN was produced from A. niger. To validate these results, products obtained via fungal degradation were compared to products obtained from the degradation of two chitin polymers. The observed retention times and mass spectral extractions provided a two-step validation confirming that GlcN was produced from fungal-derived chitin. Our studies qualitatively illustrate that GlcN can be produced from A. niger; applying these methods to a more diverse range of fungi offers the potential to render a broad screening method for fungal detection pertinent to diagnosis of fungal infections.Graphical abstract
Highlights
Aspergillus niger is a ubiquitous species of filamentous fungi [1, 2]
We propose that liquid chromatographymass spectrometry (LC-MS) can be used as a broad screening method to assist in diagnosing pulmonary fungal infections (e.g., Chronic pulmonary aspergillosis (CPA)) via the detection of chitin contained within fungal cell walls
LC-MS analysis indicated the presence of a chromatographic peak with a retention time matching that of GlcN produced from the degradation of chitin
Summary
Aspergillus niger is a ubiquitous species of filamentous fungi [1, 2]. Colony formation can be visualized after a few days of growth by characteristic black coloration, which is produced by conidial spores [3]. A. niger is a species of filamentous fungus which produces branching hyphae [4] and is widely studied for its ability to produce citric acid, along with industrially significant enzymes such as glucoamylase [5,6,7]. It is a well-known plant pathogen [8], and is responsible for black mold formation which can lead to food spoilage in human food sources such as onions and grapes [5]. These methods hold the potential to screen for the presence of filamentous fungi, and play a role in fungal infection diagnostics
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