Fruit softening in Fragaria (strawberry) is proposed to be associated with the modification of cell wall components such as xyloglucan by the action of cell wall-modifying enzymes. This study focuses on the in vitro and in vivo characterization of two recombinant xyloglucan endotransglucosylase/hydrolases (XTHs) from Fragaria vesca, FvXTH9 and FvXTH6. Mining of the publicly available F. vesca genome sequence yielded 28 putative XTH genes. FvXTH9 showed the highest expression level of all FvXTHs in a fruit transcriptome data set and was selected with the closely related FvXTH6 for further analysis. To investigate their role in fruit ripening in more detail, the coding sequences of FvXTH9 and FvXTH6 were cloned into the vector pYES2 and expressed in Saccharomyces cerevisiae. FvXTH9 and FvXTH6 displayed xyloglucan endotransglucosylase (XET) activity towards various acceptor substrates using xyloglucan as the donor substrate. Interestingly, FvXTH9 showed activity of mixed-linkage glucan:xyloglucan endotransglucosylase (MXE) and cellulose:xyloglucan endotransglucosylase (CXE). The optimum pH of both FvXTH9 and FvXTH6 was 6.5. The prediction of subcellular localization suggested localization to the secretory pathway, which was confirmed by localization studies in Nicotiana tabacum. Overexpression showed that Fragaria 3 ananassa fruits infiltrated with FvXTH9 and FvXTH6 ripened faster and showed decreased firmness compared with the empty vector control pBI121. Thus FvXTH9 and also FvXTH6 might promote strawberry fruit ripening by the modification of cell wall components.

Recently, the architectural and physical properties of the fungus comb from subterranean termite Macrotermes gilvus Hagen (Isoptera: Termitidae) mounds had been studied and it is important to determine its chemical profile as well as to evaluate its anti-staining-fungi activity. The results showed that fungus comb of M. gilvus has a high crude ash (30.57%), fiber (25.46%), starch (7.76%), protein (5.80%, 5.53% amino acid), acid-insoluble ash (3.45%), and fat (0.73%). It also contained phenol hydroquinone, steroids, terpenoids, and saponin compounds. Seventeen amino acids were identified via high-performance liquid chromatography analysis, of which arginine, leucine, glutamate, and aspartic acid were the majority. According to gas chromatography-mass spectrometry analysis, the n-hexane extract consists of several types of fatty acid derivatives. Meanwhile, the ethyl acetate (EtOAc) extracts were primarily phenol groups with 1,2,3-propanetriol (glycerol) at the highest relative concentration. Four fungus-comb extracts (n-hexane, EtOAc, MeOH, and water) inhibited the Aspergillus foetidus fungus, with inhibition rates ranging from 24.17% to 100% and EtOAc extract as the most active extract. It appears that EtOAc extracts from the M. gilvus fungus comb can be considered an active ingredient source of novel organic fungicide in preventing wood-staining fungi attacks on susceptible wood.

Cocoa fermentation is an essential process that produces flavor precursors. However, many small farmers in Indonesia directly dry their cocoa beans without fermentation due to low yield and long fermentation time, resulting in fewer flavor precursors and cocoa flavor. Therefore, this study aimed to enhance the flavor precursors, particularly free amino acids and volatile compounds, of unfermented cocoa beans by hydrolysis, using bromelain. Unfermented cocoa beans were previously hydrolyzed with bromelain at concentrations of 3.5, 7, and 10.5 U/mL for 4, 6, and 8 h, respectively. An analysis of enzyme activity, degree of hydrolysis, free amino acids, reducing sugar, polyphenols, and volatile compounds was then conducted using unfermented and fermented cocoa beans as negative and positive controls, respectively. The results showed that the highest degree of hydrolysis was 42.95% at 10.5 U/mL for 6 h, although it was not significantly different from the hydrolysis at 3.5 U/mL for 8 h. This indicates a higher reducing sugar and lower polyphenols content than unfermented cocoa beans. There was also an increase in free amino acids, especially hydrophobic amino acids, such as phenylalanine, valine, leucine, alanine, and tyrosine, and desirable volatile compounds, such as pyrazines. Therefore, this suggests that hydrolysis with bromelain increased the flavor precursors and cocoa-bean flavors.

Thermostable proteases that optimally withstand the high‐temperature conditions of thermophilic bacteria could be produced and purified, which would be highly beneficial for use in industry. Geobacillus sp. is a thermophilic bacterium that can be found in various environmental conditions. The goal of this study was to isolate and characterize thermostable serine protease that had been produced by thermophilic Geobacillus sp. strain DS3. The proteolytic index was measured in a solid medium. The expression of protease was optimized by Geobacillus sp. DS3 at 50 °C for 18 h. Targeted protease was purified using ammonium sulfate (40‐60%) and DEAE Sephadex A‐25 resin. Using SDS‐PAGE, the molecular weight of the enzyme was predicted to be around 32 kDa. Purified thermostable protease was highly activated at 70 °C, pH 9.6 stable for 1 h, and inhibited by PMSF. Therefore, this enzyme is classified as a thermostable alkaline serine protease. Its kinetic study revealed specific activity of 0.41 U/mg (Vmax) and 0.25 mg/mL (KM). Overall, a thermostable alkaline serine protease from Geobacillus sp. DS3 showed high activity at high temperatures and alkaline pH, which is vital for application in industries such as leather processing and detergent formulation.