Shotgun proteomic analysis of tiger milk mushroom (Lignosus rhinocerotis) and the isolation of a cytotoxic fungal serine protease from its sclerotium

Yap, Hui-Yeng Y.; Fung, Shin-Yee; Ng, Szu-Ting; Tan, Cher Siang; Tan, Nget-Hong

doi:10.1016/j.jep.2015.08.042

Cited by 44 publications

(44 citation statements)

References 50 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…In particular, its anticancer activities have been well explored by several research groups [9, 51, 52]. A partially purified fraction made up of two putative cytotoxic subtilisin-like fungal serine proteases has been recently reported to exhibit apoptosis-inducing activity in MCF7 cells [9] and more recently, Pushparajah et al [52] synthesised a functional recombinant lectin-like fungal immunomodulatory protein using cDNA of L. rhinocerotis , which exhibit cytotoxicity against cancer cell lines. While the reported bioactive compounds are of high molecular weight protein molecules, here, we characterised three STSs from L. rhinocerotis using yeast heterologous system, which led to the discovery of (+)-torreyol that exhibited potent cytotoxicity against MCF7 and α-cadinol that is less active.…”

Section: Discussionmentioning

confidence: 99%

“…The genome and transcriptome studies of L. rhinocerotis have provided insights into the biology of this medicinal mushroom and opened up new opportunities for exploration via genomic approaches [6–9]. Transcriptomics analysis revealed genes encoding several small secreted cysteine-rich proteins (cerato-platanins and hydrophobins) and lectins were among the highest expressed genes in the sclerotium of L. rhinocerotis and may have important implications for various bioactivities relevant to human diseases [7].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Heterologous expression of cytotoxic sesquiterpenoids from the medicinal mushroom Lignosus rhinocerotis in yeast

et al. 2017

Self Cite

View full text Add to dashboard Cite

BackgroundGenome mining facilitated by heterologous systems is an emerging approach to access the chemical diversity encoded in basidiomycete genomes. In this study, three sesquiterpene synthase genes, GME3634, GME3638, and GME9210, which were highly expressed in the sclerotium of the medicinal mushroom Lignosus rhinocerotis, were cloned and heterologously expressed in a yeast system.ResultsMetabolite profile analysis of the yeast culture extracts by GC–MS showed the production of several sesquiterpene alcohols (C15H26O), including cadinols and germacrene D-4-ol as major products. Other detected sesquiterpenes include selina-6-en-4-ol, β-elemene, β-cubebene, and cedrene. Two purified major compounds namely (+)-torreyol and α-cadinol synthesised by GME3638 and GME3634 respectively, are stereoisomers and their chemical structures were confirmed by 1H and 13C NMR. Phylogenetic analysis revealed that GME3638 and GME3634 are a pair of orthologues, and are grouped together with terpene synthases that synthesise cadinenes and related sesquiterpenes. (+)-Torreyol and α-cadinol were tested against a panel of human cancer cell lines and the latter was found to exhibit selective potent cytotoxicity in breast adenocarcinoma cells (MCF7) with IC50 value of 3.5 ± 0.58 μg/ml while α-cadinol is less active (IC50 = 18.0 ± 3.27 μg/ml).ConclusionsThis demonstrates that yeast-based genome mining, guided by transcriptomics, is a promising approach for uncovering bioactive compounds from medicinal mushrooms.Electronic supplementary materialThe online version of this article (doi:10.1186/s12934-017-0713-x) contains supplementary material, which is available to authorized users.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Heterologous expression of cytotoxic sesquiterpenoids from the medicinal mushroom Lignosus rhinocerotis in yeast

et al. 2017

Self Cite

View full text Add to dashboard Cite

show abstract

“…Solvent‐free plugs were digested using Pierce™ Trypsin Protease (Thermo Scientific, MA) and proteins retrieved ZipTip C18 cartridge column pipette tip (Merck Millipore) according to the respective manufacturers' instruction. The peptides were separated using Agilent 1200HPLC‐Chip/MS Interface, coupled with Agilent 6520 Accurate‐Mass Q‐TOF LC/MS (Agilent Technologies, CA) and analyzed according to parameters used previously . The peptide sample was redissolved in Solution A (0.1% formic acid) and loaded onto the 160 ηL enrichment column (C18, 300 Å) of microfluidic nanospray chip at 4 μL/min.…”

Section: Methodsmentioning

confidence: 99%

“…Grifola frondosa lectin (GFL) was cytotoxic against HeLa cells . Selective cytotoxicity toward cancer cells without harming normal cells was demonstrated in mushroom properties from Agaricus blazei , Clitocybe nebularis , Coprinus comatus , Ganoderma lucidum , and Lignosus rhinocerus …”

Section: Introductionmentioning

confidence: 99%

Cloning, overexpression, purification, and modeling of a lectin (Rhinocelectin) with antiproliferative activity from Tiger Milk Mushroom, Lignosus rhinocerus

et al. 2019

View full text Add to dashboard Cite

A lectin gene from the Tiger Milk Mushroom Lignosus rhinocerus TM02® was successfully cloned and expressed via vector pET28a in Escherichia coli BL21(DE3). The recombinant lectin, Rhinocelectin, with a predicted molecular mass of 22.8 kDa, was overexpressed in water‐soluble form without signal peptide and purified via native affinity chromatography Ni‐NTA agarose. Blast protein analysis indicated the lectin to be homologous to jacalin‐related plant lectin. In its native form, Rhinocelectin exists as a homo‐tetramer predicted with four chains of identical proteins consisting of 11 beta‐sheet structures with only one alpha‐helix structure. The antiproliferative activity of the Rhinocelectin against human cancer cell lines was concentration dependent and selective. The IC50 values against triple negative breast cancer cell lines MDA‐MB‐231 and breast cancer MCF‐7 are 36.52 ± 13.55 μg mL−1 and 53.11 ± 22.30 μg mL−1, respectively. Rhinocelectin is only mildly cytotoxic against the corresponding human nontumorigenic breast cell line 184B5 with IC50 value at 142.19 ± 36.34 μg mL−1. The IC50 against human lung cancer cell line A549 cells is 46.14 ± 7.42 μg mL−1 while against nontumorigenic lung cell line NL20 is 41.33 ± 7.43 μg mL−1. The standard anticancer drug, Doxorubicin exhibited IC50 values mostly below 1 μg mL−1 for the cell lines tested. Flow cytometry analysis showed the treated breast cancer cells were arrested at G0/G1 phase and apoptosis induced. Rhinocelectin agglutinated rat and rabbit erythrocytes at a minimal concentration of 3.125 μg mL−1 and 6.250 μg mL−1, respectively.

show abstract

“…Its protease and cytotoxic activities were attenuated in the presence of phenylmethylsulfonyl fluoride, indicating a relationship between the two activities (Yap et al 2015).…”

Section: Fungal Proteins and Peptides With Antitumor Activitymentioning

confidence: 99%

Fungal proteinaceous compounds with multiple biological activities

Cheung

Wong

et al. 2016

Appl Microbiol Biotechnol

View full text Add to dashboard Cite

Fungi comprise organisms like molds, yeasts and mushrooms. They have been used as food or medicine for a long time. A large number of fungal proteins or peptides with diverse biological activities are considered as antibacterial, antifungal, antiviral and anticancer agents. They encompass proteases, ribosome inactivating proteins, defensins, hemolysins, lectins, laccases, ribonucleases, immunomodulatory proteins, and polysaccharopeptides. The target of the present review is to update the status of the various bioactivities of these fungal proteins and peptides and discuss their therapeutic potential.

show abstract

Shotgun proteomic analysis of tiger milk mushroom (Lignosus rhinocerotis) and the isolation of a cytotoxic fungal serine protease from its sclerotium

Cited by 44 publications

References 50 publications

Heterologous expression of cytotoxic sesquiterpenoids from the medicinal mushroom Lignosus rhinocerotis in yeast

Heterologous expression of cytotoxic sesquiterpenoids from the medicinal mushroom Lignosus rhinocerotis in yeast

Cloning, overexpression, purification, and modeling of a lectin (Rhinocelectin) with antiproliferative activity from Tiger Milk Mushroom, Lignosus rhinocerus

Fungal proteinaceous compounds with multiple biological activities

Contact Info

Product

Resources

About