BackgroundTermitomyces heimii is a basidiomycete fungus that has a symbiotic relationship with termites, and it is an edible mushroom with a unique flavour and texture. T. heimii is also one of the most difficult mushrooms to cultivate throughout the world. Little is known about the growth and development of these mushrooms, and the available information is insufficient or poor. The purpose of this study was to provide a base of knowledge regarding the biological processes involved in the development of T. heimii. The proteomic method of 2 dimensional difference gel electrophoresis 2D-DIGE was used to determine and examine the protein profiles of each developmental stage (mycelium, primordium and fruiting body). Total proteins were extracted by TCA-acetone precipitation.ResultsA total of 271 protein spots were detected by electrophoresis covering pH 3–10 and 10–250 kDa. Selected protein spots were subjected to mass spectrometric analyses with matrix-assisted laser desorption/ionisation (MALDI TOF/TOF). Nineteen protein spots were identified based on peptide mass fingerprinting by matching peptide fragments to the NCBI non-redundant database using MASCOT software. The 19 protein spots were categorised into four major groups through KEGG pathway analysis, as follows: carbohydrate metabolism, energy metabolism, amino acid metabolism and response to environmental stress.ConclusionsThe results from our study show that there is a clear correlation between the changes in protein expression that occur during different developmental stages. Enzymes related to cell wall synthesis were most highly expressed during fruiting body formation compared to the mycelium and primordial stages. Moreover, enzymes involved in cell wall component degradation were up-regulated in the earlier stages of mushroom development.

Tiger's milk mushroom is known for its valuable medicinal properties, especially the tuber part. However, wild tuber is very hard to obtain as it grows underground. This study first aimed to cultivate tiger's milk mushroom tuber through a cultivation technique, and second to compare nutritional and mycochemical contents, antioxidant and cytotoxic activities and compound screening of the cultivated tuber with the wild tuber. Results showed an increase in carbohydrate content by 45.81% and protein content by 123.68% in the cultivated tuber while fat content reduced by 13.04%. Cultivated tuber also showed an increase of up to 64.21% for total flavonoid-like compounds and 62.51% of total β-D-glucan compared to the wild tuber. The antioxidant activity of cultivated tuber and wild tuber was 760 and 840 µg mL, respectively. The cytotoxic activity of boiled water extract of cultivated tuber against a human lung cancer cell line (A549) was 65.50 ± 2.12 µg mL and against a human breast cancer cell line (MCF7) was 19.35 ± 0.11 µg mL. β-D-glucan extract from the purification of boiled water extract of cultivated tuber showed cytotoxic activity at 57.78 ± 2.29 µg mL against A549 and 33.50 ± 1.41 µg mL against MCF7. However, the β-glucan extract from wild tuber did not show a cytotoxic effect against either the A549 or MCF7 cell lines. Also, neither of the extracts from cultivated tuber and wild tuber showed an effect against a normal cell line (MRC5). Compound profiling through by liquid chromatography mass spectrometry (LC/MS) showed the appearance of new compounds in the cultivated tuber. In conclusion, our cultivated tuber of tiger's milk mushroom using a new recipe cultivation technique showed improved nutrient and bioactive compound contents, and antioxidant and cytotoxic activities compared to the wild tuber. Further investigations are required to obtain a better quality of cultivated tuber.

Coffee is one of the most preferred beverages due to its flavour and aroma, which is partially contributed by the presence of caffeine. However, there are many negative effects of caffeine on human health. Alternative products like date seed and barley beverage have become an interest to switch from caffeine to caffeine-free beverages. The present work thus aimed to evaluate the nutritional properties and toxicity of date seed and barley powders as compared to Arabica coffee powder. Samples were analysed for its caffeine content, antioxidant activity, and toxicity activity from boiled water extract, whereas the nutrition compositions and heavy metal contents were analysed based on respective extraction method performed. A mass spectral peak of caffeine was detected in the Arabica coffee but not in the date seed and barley powders. All three samples were shown to possess antioxidant activities with Arabica coffee yielding the highest. Arabica coffee, however, exhibited a moderate level of toxicity to human lung fibroblast (MRC-5) cell line with IC50 of 230 ± 40 µg/mL at the extract concentration. There was no inhibition on 50% MRC-5 cell viability showed by the date seed and barley powders up to 10 mg/mL extract concentration. The abundance of heavy metals detected in all samples was lower than the regulatory limits. Our findings therefore further supported the advantages of date seed and barley powders as alternatives to coffee beverage as both contained undetected amount of caffeine, low fat and high carbohydrate contents, and possessed good antioxidant activity with low potential health risks.