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Shotgun proteomics approaches for authentication, biological analyses, and allergen detection in feed and food -grade insect species

ABSTRACT: Untargeted proteomics approaches were shown to be suitable for composition and authenticity analyses of highly processed mixed food and feed products. In this work, a normal-flow tandem mass spectrometry-based proteomics method was setup for analysis and authentication of insect meal from five different species. Novel data acquired on a Q Exactive Orbitrap (QE) was compared with previously published data obtained on QTOF. Data from both proteomics workflows were compared using compareMS2 and a Trans-Proteomics Pipeline (TPP). The results obtained for species differentiation, peptides, and protein markers detection were comparable across both approaches. The collected mass spectrometry data from both instruments also were used to build spectral libraries for insect species and matching was performed successfully. Lastly, both datasets were screened for known allergens and arginine kinase and tropomyosin were consistently detected confirming the presence of potential allergenic risks associated with the consumption of edible insects.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Alphitobius Diaperinus (ncbitaxon:27448) Hermetia Illucens (ncbitaxon:343691) Zophobas Morio Acheta Domesticus (ncbitaxon:6997) Tenebrio Molitor (ncbitaxon:7067)

SUBMITTER: Josef Daniel Rasinger

PROVIDER: MSV000088034 | MassIVE | Mon Aug 30 04:06:00 BST 2021

REPOSITORIES: MassIVE

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Project description:Arabidopsis thaliana plants were infested i) with sucking insect herbivores (the generalist aphid Myzus persicae and the specialist aphid Brevicoryne brassicae), ii) with chewing insect herbivores (generalist caterpillars of Spodoptera exigua and specialist caterpillars of Pieris rapae) or iii) were treated by wounding. For each treatment, rosette leaves were harvested at two time points (6h and 24h) after removal of insects. For chewing herbivores and wounding both local, i.e. immediately damaged leaves, and systemic, i.e. undamaged leaves from the same plant, were collected. Control plants were uninfested, but otherwise equally treated and harvested in parallel. We tested the hypothesis that Arabidopsis can recognize and respond differentially to insect species at the transcriptional level using a genome wide microarray. Transcriptional reprogramming was characterized using co-expression analysis in damaged and undamaged leaves at two times in response to mechanical wounding and four insect species. In all, 2778 (10.6%) of annotated genes on the array were differentially expressed in at least one treatment. Responses differed mainly between aphid and caterpillar and sampling times. Responses to aphids and caterpillars shared only 10% of up-regulated and 8% of down-regulated genes. Responses to two caterpillars shared 21% and 12% of up- and down-regulated genes, whereas responses to the two aphids shared only 7% and 4% of up-regulated and down-regulated genes. Overlap in genes expressed between 6h and 24h was 3-15%, and depended on the insect species. Responses in attacked and unattacked leaves differed at 6h but converged by 24h. Genes responding to the insects are also responsive to many stressors and included primary metabolism. Aphids down-regulated amino acid catabolism; caterpillars stimulated production of amino acids involved in glucosinolate synthesis. Co-expression analysis revealed 17 response networks. Transcription factors were a major portion of differentially expressed genes throughout and responsive genes shared most of the known or postulated binding sites.

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Shotgun proteomics approaches for authentication, biological analyses, and allergen detection in feed and food -grade insect species

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