To retrieve a local isolate, a loopful of spores was scraped from moldy bread and cultured on potato dextrose agar (PDA). The PDA contained potato extract (4.0 g) (fresh, unpeeled potatoes), glucose (20 g) (Merck KGaA, Darmstadt, Germany), and agar (20 g) (Oxoid, Basingstoke, UK) in distilled water (1000 mL) [11 (link)]. The agar was dissolved by boiling the mixture, followed by sterilization before pouring it onto plates. Sabouraud dextrose agar (SDA) (HiMedia, Mumbai, India) was also used for the parallel culturing of the isolate. Both media were incubated for five to seven days at 25 ± 2 °C [12 (link)]. Then, a cork borer was used to inoculate a fungal disk on Aspergillus differentiation agar. This agar was prepared by adding 10 g of yeast extract (HiMedia, Thane, India), 15 g of tryptone (HiMedia), 0.5 g of ferric citrate (Merck KGaA), and 15 g of agar (Oxoid) to distilled water (1000 mL). The media were boiled and autoclaved, and the inoculated plates were incubated for five to seven days at 25 ± 2 °C [13 (link)].
Potato extract
Manufactured by Merck Group
Sourced in Germany
Potato extract is a laboratory product that is derived from potatoes. It contains various organic compounds and nutrients found naturally in potatoes. The core function of potato extract is to serve as a source of these compounds for research and analysis purposes. However, a detailed description of its specific properties and applications would require more information than can be provided in a concise, unbiased manner.
Automatically generated - may contain errors
Lab products found in correlation
Dextrose, by Merck Group (3 mentions)
Glucose, by Merck Group (2 mentions)
Agar, by Merck Group (2 mentions)
Agar, by AppliChem (2 mentions)
Xevo g2 xs qtof mass spectrometer, by Waters Corporation (2 mentions)
Agar, by Thermo Fisher Scientific (1 mentions)
Tryptone, by HiMedia (1 mentions)
Ferric citrate, by Merck Group (1 mentions)
Sabouraud dextrose agar, by HiMedia (1 mentions)
Yeast extract, by HiMedia (1 mentions)
Malt extract, by Merck Group (1 mentions)
Yeast extract, by Merck Group (1 mentions)
Acquity uplc 1 class system, by Waters Corporation (1 mentions)
Arium water purification system, by Sartorius (1 mentions)
Casein hydrolysate, by Carl Roth (1 mentions)
Yeast extract, by BD (1 mentions)
Etoac, by Avantor (1 mentions)
Uplc 1 class system, by Waters Corporation (1 mentions)
Acetonitrile, by Biosolve (1 mentions)
Uplc grade methanol, by Biosolve (1 mentions)
6 protocols using potato extract
1
Isolation and Culturing of Fungal Isolates
2
Cultivation Protocols for Fungal Fruiting
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According to the identification of conidia, phialides and colony coloration, the isolate cultures were grown on YMG media, composed of 4 g/l yeast extract (Sigma-Aldrich, Germany), 10 g/l malt extract (Sigma-Aldrich, Germany), 4 g/l glucose (Sigma-Aldrich, Germany), and incubated at 20 °C for a period of 20 days with PDA media (potato extract 4 g/l, dextrose 20 g/l, agar 15 g/l; Merck, Germany).
For fruit body induction, cultures were grown on millet substrate (millet/silkworm pupae powder = 20:1 (w/w)) and brown rice substrate (brown rice/silkworm pupae powder = 20:1 (w/w)) at 20 °C under 12 h light and 12 h darkness with relative humidity of over 90%.
For fruit body induction, cultures were grown on millet substrate (millet/silkworm pupae powder = 20:1 (w/w)) and brown rice substrate (brown rice/silkworm pupae powder = 20:1 (w/w)) at 20 °C under 12 h light and 12 h darkness with relative humidity of over 90%.
3
Tandem Mass Spectrometry of Fungal Metabolites
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Tandem mass (MS/MS) spectrometry data were recorded on a Waters Xevo G2-XS QToF Mass Spectrometer (Waters®, Milford, MA, USA) connected to a Waters Acquity I-Class UPLC system (Waters®, Milford, MA, USA) in positive mode. The organic solvent used for MS/MS analysis was ULC/MS grade. An in-house Arium® Water Purification System (Sartorius, Goettingen, Germany) was used for the preparation of milli Q water. Potato extract and dextrose used for fungal cultivation were purchased from Sigma-Aldrich (Schnelldorf, Germany) and Merck (Darmstadt, Germany), respectively. Agar was purchased from Applichem (Darmstadt, Germany).
4
Fungal Extraction and UPLC-QToF-MS/MS Analysis
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EtOAc (used for fungal extraction) was purchased from VWR International GmbH, Hannover, Germany. UPLC grade methanol, acetonitrile and water used for UPLC/MS analysis were purchased from BiosolveChimie, Dieuze, France. Formic acid (UPLC/MS optigrade) was obtained from LGC Standards Promochem©, Wesel, Germany. UPLC-QToF-MS/MS analyses were carried out on an ACQUITY UPLC I-Class System coupled to the Xevo G2-XS QToF Mass Spectrometer (Waters®, Milford, MA, USA). Czapek broth, yeast extracts and malt extracts were purchased from BD Bioscience, Sparks, NE, USA. Agar was purchased from Applichem, Darmstadt, Germany. Peptone from soymeal and glucose were purchased from Merck, Darmstadt, Germany. Potato extract was from Sigma-Aldrich, Schnelldorf, Germany. Sucrose was purchased from Handelsmarken, Offenburg, Germany. Casein hydrolysate was purchased from Carl Roth, Karlsruhe, Germany.
5
Cultivation of Ganoderma lucidum Mycelium
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The mycelium mother culture of G. lucidum (M 9720) was purchased from Mycelia bvba (Nevele, Belgium). The mycelium mother culture was then maintained on potato dextrose agar (PDA) slopes, incubated at 28 °C for up to 7 days and stored at 4 °C for subsequent subculturing. PDA media was made from the following components in grams per litre: potato extract, 4 g; dextrose, 20 g; and agar, 15 g. Individual components were purchased from Sigma Aldrich, St. Louis, Mo.
EFB fibres were collected from the Heng Huat Group in Malaysia, and sawdust from Albizia chinensis, a common tropical plant, was collected from a San Ho timber workshop in Singapore. Mycelium growing substrates were supplemented with 10% (W/W) wheat bran (Bob’s Red Mill, Product of USA) for additional nutrition and 2% (W/W) calcium carbonate (CaCO3) (LushGro, Singapore) to adjust the mixture pH level. Some of the DMC samples were coated with an oil-based coating from Osmo Holz und Color (Warendorf, Germany) for weathering tests. Verification of the mycelium species and plant fibres used in this research was carried out with companies that handed over the respective materials for the research. Necessary documents were provided to the Singapore custom office to ensure complying with Singapore laws in terms of imported live specimens.
EFB fibres were collected from the Heng Huat Group in Malaysia, and sawdust from Albizia chinensis, a common tropical plant, was collected from a San Ho timber workshop in Singapore. Mycelium growing substrates were supplemented with 10% (W/W) wheat bran (Bob’s Red Mill, Product of USA) for additional nutrition and 2% (W/W) calcium carbonate (CaCO3) (LushGro, Singapore) to adjust the mixture pH level. Some of the DMC samples were coated with an oil-based coating from Osmo Holz und Color (Warendorf, Germany) for weathering tests. Verification of the mycelium species and plant fibres used in this research was carried out with companies that handed over the respective materials for the research. Necessary documents were provided to the Singapore custom office to ensure complying with Singapore laws in terms of imported live specimens.
6
Filamentous Fungal Strain Cultivation
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Five edible filamentous fungal strains (Rhizopus oryzae CCUG 28958, R. microsporus var. oligosporus CBS 112586, R. oryzae var. delemar CBS 145940, Aspergillus oryzae var. oryzae CBS 819.72, Neurospora intermedia CBS 131.92) were used. The fungal strains were maintained on Potato Dextrose agar (PDA; pH 5.6, containing 4 g/L potato extract (Sigma-Aldrich, Buchs, Switzerland), 20 g/L glucose (Fisher Scientific, Loughborough, UK) and 15 g/L agar (Sigma, Buchs, Switzerland)). For fungal inoculation, pre-grown plates were treated with 20 mL of distilled water, and then 100 μL of spore suspension were inoculated into fresh PDA using a L-shape sterile disposable plastic spreader. The fungal strains were cultured at 30 • C for three days and then stored at 4 • C until use for a maximum of one month.
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