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iTRAQ-facilitated proteomic analysis of Bacillus cereus via degradation of malachite green
Bobo Wang , Jing Lu , Junfang Zheng , Zhisheng Yu
J. Microbiol. 2021;59(2):142-150.   Published online February 1, 2021
DOI: https://doi.org/10.1007/s12275-021-0441-0
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AbstractAbstract
The wide use of malachite green (MG) as a dye has caused substantial concern owing to its toxicity. Bacillus cereus can against the toxic effect of MG and efficiently decolourise it. However, detailed information regarding its underlying adaptation and degradation mechanisms based on proteomic data is scarce. In this study, the isobaric tags for relative and absolute quantitation (iTRAQ)-facilitated quantitative method was applied to analyse the molecular mechanisms by which B. cereus degrades MG. Based on this analysis, 209 upregulated proteins and 198 downregulated proteins were identified with a false discovery rate of 1% or less during MG biodegradation. Gene ontology and KEGG analysis determined that the differentially expressed proteins were enriched in metabolic processes, catalytic activity, antioxidant activity, and responses to stimuli. Furthermore, real-time qPCR was utilised to further confirm the regulated proteins involved in benzoate degradation. The proteins BCE_4076 (Acetyl-CoA acetyltransferase), BCE_5143 (Acetyl-CoA acetyltransferase), BCE_5144 (3-hydroxyacyl-CoA dehydrogenase), BCE_4651 (Enoyl-CoA hydratase), and BCE_5474 (3-hydroxyacyl-CoA dehydrogenase) involved in the benzoate degradation pathway may play an important role in the biodegradation of MG by B. cereus. The results of this study not only provide a comprehensive view of proteomic changes in B. cereus upon MG loading but also shed light on the mechanism underlying MG biodegradation by B. cereus.

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  • Engineering globins for efficient biodegradation of malachite green: two case studies of myoglobin and neuroglobin
    Jiao Liu, Jia-Kun Xu, Hong Yuan, Xiao-Juan Wang, Shu-Qin Gao, Ge-Bo Wen, Xiang-Shi Tan, Ying-Wu Lin
    RSC Advances.2022; 12(29): 18654.     CrossRef
The putative polysaccharide synthase AfCps1 regulates Aspergillus fumigatus morphogenesis and conidia immune response in mouse bone marrow-derived macrophages
Sha Wang , Anjie Yuan , Liping Zeng , Sikai Hou , Meng Wang , Lei Li , Zhendong Cai , Guowei Zhong
J. Microbiol. 2021;59(1):64-75.   Published online November 17, 2020
DOI: https://doi.org/10.1007/s12275-021-0347-x
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AbstractAbstract
Aspergillus fumigatus is a well-known opportunistic pathogen that causes invasive aspergillosis (IA) infections with high mortality in immunosuppressed individuals. Morphogenesis, including hyphal growth, conidiation, and cell wall biosynthesis is crucial in A. fumigatus pathogenesis. Based on a previous random insertional mutagenesis library, we identified the putative polysaccharide synthase gene Afcps1 and its paralog Afcps2. Homologs of the cps gene are commonly found in the genomes of most fungal and some bacterial pathogens. Afcps1/cpsA is important in sporulation, cell wall composition, and virulence. However, the precise regulation patterns of cell wall integrity by Afcps1/cpsA and further effects on the immune response are poorly understood. Specifically, our in-depth study revealed that Afcps1 affects cell-wall stability, showing an increased resistance of ΔAfcps1 to the chitinmicrofibril destabilizing compound calcofluor white (CFW) and susceptibility of ΔAfcps1 to the β-(1,3)-glucan synthase inhibitor echinocandin caspofungin (CS). Additionally, deletion of Afcps2 had a normal sporulation phenotype but caused hypersensitivity to Na+ stress, CFW, and Congo red (CR). Specifically, quantitative analysis of cell wall composition using high-performance anion exchange chromatography- pulsed amperometric detector (HPAEC-PAD) analysis revealed that depletion of Afcps1 reduced cell wall glucan and chitin contents, which was consistent with the downregulation of expression of the corresponding biosynthesis genes. Moreover, an elevated immune response stimulated by conidia of the ΔAfcps1 mutant in marrow-derived macrophages (BMMs) during phagocytosis was observed. Thus, our study provided new insights into the function of polysaccharide synthase Cps1, which is necessary for the maintenance of cell wall stability and the adaptation of conidia to the immune response of macrophages in A. fumigatus.

Citations

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  • Study on the metabolic changes and regulatory mechanism of Aspergillus flavus conidia germination
    Sifan Jia, Chong Li, Yu An, Desheng Qi, Erik F. Y. Hom
    Microbiology Spectrum.2024;[Epub]     CrossRef
  • Chitin Biosynthesis in Aspergillus Species
    Veronica S. Brauer, André M. Pessoni, Mateus S. Freitas, Marinaldo P. Cavalcanti-Neto, Laure N. A. Ries, Fausto Almeida
    Journal of Fungi.2023; 9(1): 89.     CrossRef
  • Evidencing New Roles for the Glycosyl-Transferase Cps1 in the Phytopathogenic Fungus Botrytis cinerea
    Matthieu Blandenet, Isabelle R. Gonçalves, Christine Rascle, Jean-William Dupuy, François-Xavier Gillet, Nathalie Poussereau, Mathias Choquer, Christophe Bruel
    Journal of Fungi.2022; 8(9): 899.     CrossRef

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