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Cysteine-rich peptide from the gigantic edible mushroom Kusaghiporia usambarensis (Laetiporaceae)
Systematisk biologi, Systematic Biology. (Tibell)ORCID iD: 0000-0003-3442-9262
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Biochemistry and Microbiology.ORCID iD: 0000-0003-4154-2378
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Organismal Biology, Systematic Biology.ORCID iD: 0000-0003-4143-9856
University of Dar es Salaam, Department of molecular Biology and Biotechnology.ORCID iD: 0000-0002-3105-8374
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(English)Manuscript (preprint) (Other academic)
Abstract [en]

Cysteine-rich peptides are produced by various organisms across all kingdoms and have triggered an interest in isolation of molecules for novel drug development. In this study, we report a novel cysteine-rich peptide, kusaghitide, isolated from the gigantic medicinal mushroom Kusaghiporia usambarensis. It is highly expressed in the K. usambarensis transcriptome and it is the most abundant compound in the methanol-water extract. The 54 amino acid residue long peptide was isolated through aqueous methanol 50% and a sample was reduced, alkylated and cleaved enzymatically. De novo sequencing was done by LC-MS/MS and obtained sequences were used for mining the transcriptome to search for the complete gene. The peptide was recombinantly expressed in One Shot BL21 Star Escherichia coli using lysogenic broth and minimal media. Its 3D NMR structure was determined using 2D and 3D NMR. Three hypothetical protein sequences similar to kusaghitide originate from Laetiporus sulphureusWolfiporia cocos and Sparassis crispa with per cent similarity of 76% and 58% and 53% respectively and were found by BLAST search in the NCBI database. Kusaghitide did not inhibit the growth of either Escherichia coli or Staphylococcus aureus. This is first report of a peptide from K. usambarensis in Laetiporaceae.

Keywords [en]
Kusaghiporia, medicinal mushroom, recombinant protein production, cysteine-rich peptides, peptide structure.
National Category
Natural Sciences
Identifiers
URN: urn:nbn:se:uu:diva-405387OAI: oai:DiVA.org:uu-405387DiVA, id: diva2:1399540
Available from: 2020-02-28 Created: 2020-02-28 Last updated: 2020-02-28
In thesis
1. Polyporoid fungi of Tanzania: Taxonomy, transcriptomics and biochemical analyses of Kusaghiporia usambarensis and Piptoporellus baudonii
Open this publication in new window or tab >>Polyporoid fungi of Tanzania: Taxonomy, transcriptomics and biochemical analyses of Kusaghiporia usambarensis and Piptoporellus baudonii
2020 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Polyporoid fungi refers to basidiomycetes with fruiting bodies with the hymenium located to the inner surfaces of pores or narrow tubes. The majority of polyporoids belongs to Polyporales. Most Polyporales are saprobes, but some are plant pathogens. The overall aim of this thesis was to study the taxonomy, systematics and chemistry of the two species Kusaghiporia usambarensis (saprobic) and Piptoporellus baudonii (a plant pathogen) collected from Tanzania, using morphological and molecular approaches, combined with transcriptomics and pharmacognostic investigations.

The main contribution of this thesis includes the description a new genus with the new species K. usambarensis from the Usambara Mountains, Tanzania; investigation of the chemical composition of volatile compounds from this medicinal mushroom; isolation and structure determination of a novel and most abundant peptide in K. usambarensis, and further to elucidate the phylogenetic position of Piptoporellus baudonii (formerly known as Laetiporus baudonii) by using a four molecular markers dataset.

Paper I was conducted applying a classical taxonomic approach, including both morphological and phylogenetic analyses, to describe a new genus and species K. usambarensis. Paper II, investigated volatiles and volatile derivatives in dichloromethane extracts of K. usambarensis analysed by GC-MS and NMR spectroscopy. The main elements were phenols, and esters, compounds that may explain the formerly reported antioxidant activity and traditional medicinal use of the mushroom. In paper III, screening of peptides in K. usambarensis revealed a novel cysteine-rich peptide, highly expressed at gene level and the most abundant compound in the fruiting body. Combined LC-MS and transcriptome analyses were used to determine the peptide sequence, and subsequently NMR spectroscopy to determine the 3D structure of the novel peptide, kusaghitide. In paper IV molecular techniques were used to elucidate the phylogenetic position of the parasitic Laetiporus baudonii. Phylogenetic analyses of combined 5.8S, nrLSU, nrSSU, and TEF1 gene sequences placed L. baudonii in the genus Piptoporellus, hence the new combination Piptoporellus baudonii was proposed. This thesis has contributed to build capacity in the fields of mycology, systematics and pharmacognosy in order to reinforce ecological knowledge and ethnopharmaceutical research for future drug discovery in Tanzania and Africa at large.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2020. p. 56
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 1908
Keywords
Systematics, Laetiporaceae, pharmacognosy, cysteine-rich peptide, baudonii
National Category
Biological Sciences
Research subject
Biology with specialization in Systematics
Identifiers
urn:nbn:se:uu:diva-405548 (URN)978-91-513-0881-4 (ISBN)
Public defence
2020-04-17, Ekmansalen, Evolutionsbiologiskt centrum (EBC), Norbyvägen 14-18, Uppsala, 10:00 (English)
Opponent
Supervisors
Available from: 2020-03-26 Created: 2020-02-28 Last updated: 2020-03-26

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Chi, Celestine N.Jacobsson, ErikWedén, ChristinaGöransson, Ulf

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Hussein, JumaChi, Celestine N.Tibell Savić, SanjaTibuhwa, DonathaJacobsson, ErikWedén, ChristinaGöransson, Ulf
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