Isolation, screening, and characterization of the newly isolated osmotolerant yeast Wickerhamomyces anomalus BKK11-4 for the coproduction of glycerol and arabitol.

This study explored the isolation and screening of an osmotolerant yeast, Wickerhamomyces anomalus BKK11-4, which is proficient in utilizing renewable feedstocks for sugar alcohol production. In batch fermentation with high initial glucose concentrations, W. anomalus BKK11-4 exhibited notable production of glycerol and arabitol. The results of the medium optimization experiments revealed that trace elements, such as H3BO3, CuSO4, FeCl3, MnSO4, KI, H4MoNa2O4, and ZnSO4, did not increase glucose consumption or sugar alcohol production but substantially increased cell biomass. Osmotic stress, which was manipulated by varying initial glucose concentrations, influenced metabolic outcomes. Elevated glucose levels promoted glycerol and arabitol production while decreasing citric acid production. Agitation rates significantly impacted the kinetics, enhancing glucose utilization and metabolite production rates, particularly for glycerol, arabitol, and citric acid. The operational pH dictated the distribution of the end metabolites, with glycerol production slightly reduced at pH 6, while arabitol production remained unaffected. Citric acid production was observed at pH 6 and 7, and acetic acid production was observed at pH 7. Metabolomic analysis using GC/MS identified 29 metabolites, emphasizing the abundance of sugar/sugar alcohols. Heatmaps were generated to depict the variations in metabolite levels under different osmotic stress conditions, highlighting the intricate metabolic dynamics occurring post-glucose uptake, affecting pathways such as the pentose phosphate pathway and glycerolipid metabolism. These insights contribute to the optimization of W. anomalus BKK11-4 as a whole-cell factory for desirable products, demonstrating its potential applicability in sustainable sugar alcohol production from renewable feedstocks.

A Comparative Analysis of Phenolic Content, Antioxidant Activity, Antimicrobial Activity, and Chemical Profile of Coffea robusta Extracts Using Subcritical Fluid Extraction and Supercritical Carbon Dioxide Extraction.

In this study, extracts of Robusta-roasted coffee were obtained using various extraction techniques, including subcritical fluid extractions using HFC-134a and HCFC-22 under room-temperature batch extraction, frozen-temperature batch extraction, and continuous extraction conditions. Additionally, supercritical carbon dioxide (SCCO2) extraction was performed using ethanol and tetrahydrofuran as co-solvents. These extractions were performed due to the presence of potent antioxidants and antibacterial substances in the extracts. Extraction machines were built to process the extraction. The antioxidant potential of the extracts was evaluated using total phenolic content and DPPH and FRAP assays, while antibacterial potential was identified using the disk diffusion method. The results showed that HCFC-22 extraction produced the highest yield compared to other extraction methods, but HFC-134a extraction had the highest antioxidant potential values. The yield and antioxidant potential of the extracts obtained using room-temperature batch extraction were slightly higher than those obtained using frozen-temperature batch extraction and continuous extraction. The yield and antioxidant potential of the extracts obtained using SCCO2 extraction were similar to those obtained using HFC-134a and HCFC-22 extractions, and co-solvents slightly improved the extraction performance. The extracts were found to be more effective as inhibitors of Gram-positive bacteria than Gram-negative bacteria. Caffeine was the most prominent tentative chemical compound in all coffee extracts. This research study provides a better understanding of various extraction techniques using HFC-134a, HCFC-22, and SCCO2 when applied to roasted Robusta coffee beans, with a focus on yield, antioxidant potential, antimicrobial potential, and tentative chemical profiles.

Metabolomics approach to identify key volatile aromas in Thai colored rice cultivars.

In addition to white jasmine rice, Thailand has many native-colored rice varieties with numerous health benefits and the potential to become a global economic crop. However, the chemical characteristics of aromatic substances in native-colored rice are still mostly unknown. This study aimed to identify the key volatile aroma compounds and the biosynthetic pathways possibly involved in their formation in Thai native-colored rice varieties, and thus leading to the search for potential genetic markers for breeding colored rice with better aromatic properties. Twenty-three rice varieties in four categories: aromatic white, aromatic black, non-aromatic black, and non-aromatic red, were investigated (n=10 per variety). Seed husks were removed before the analysis of rice volatile aromas by static headspace gas chromatography-mass spectrometry. Untargeted metabolomics approach was used to discover the key volatile compounds in colored rice. Forty-eight compounds were detected. Thirty-eight of the 48 compounds significantly differed among groups at p<0.05, 28 of which at p<0.0001, with the non-aromatic black and red rice containing much lower content of most volatile constituents than the aromatic black and white rice. Focusing on the aromatic black rice, the samples appeared to contain high level of both compound groups of aldehydes (3-methylbutanal, 2-methylbutanal, 2-methylpropanal, pentanal, hexanal) and alcohols (butane-2,3-diol, pentan-1-ol, hexan-1-ol). Biosynthetically, these distinctive black-rice volatile compounds were proposed to be formed from the metabolic degradation of branched-chain amino acids (L-leucine, L-isoleucine and L-valine) and polyunsaturated fatty acids (linoleic acid and α-linolenic acid), involving the branched-chain aminotransferases and keto-acid decarboxylases and the 9-lipoxygonases and 13-lipoxygeases, respectively. The proposed degradative pathways of amino acids and fatty acids were well agreed with the profiles key volatile compounds detected in the Thai native-colored rice varieties.

Effect of Overfilled Solvent and Storage Time of Subcritical Extraction of Jasminum sambac on Yield, Antioxidant Activity, Antimicrobial Activity and Tentative Volatile Compounds.

Essential oil from Jasminum sambac flowers has demonstrated the potential of antioxidant and antimicrobial properties. However, jasmine flowers contain only a small amount of essential oil; therefore, subcritical fluid extraction (SFE) with HFC-134a, one of the effective extraction methods for flower extraction, was performed in this study. The percentage of overfilled solvent and storage time of the flowers were varied during the extraction. Antioxidant potential, antimicrobial potential and tentative volatile compounds were investigated in this study to observe the quality of the essential oil. It was discovered that a greater amount of overfilled solvent resulted in thicker essential oil and a longer storage time resulted in a lower amount of total oil yield. It could be seen that almost all extraction conditions did not have any significant difference in antioxidant and antimicrobial potential. The essential oil contained primary compounds such as indole, 9-tricosene, α-farnesene, muurolene, and benzyl alcohol. This study led to the conclusion that the amount of overfilled solvent from SFE affected the thickness of jasmine essential oil and its tentative volatile compounds. The longer storage time caused the significantly lower essential oil yield, but changing the extraction conditions had no significant effect on antioxidant or antimicrobial potential.

Pollinator and floral odor specificity among four synchronopatric species of Ceropegia (Apocynaceae) suggests ethological isolation that prevents reproductive interference.

Possession of flowers that trap fly pollinators is a conservative trait within the genus Ceropegia, in which pollination systems can be generalized or highly specialized. However, little is known about the role of plant-pollinator interactions in the maintenance of species boundaries. This study examined the degree of plant-pollinator specialization and identified the parameters responsible for specificity among four co-occurring Ceropegia species with overlapping flowering times. All investigated plant species were functionally specialized on pollination by Chloropidae and/or Milichiidae flies and each Ceropegia species was, in turn, ecologically highly specialized on only two pollinating fly morphospecies, though one plant species appeared more generalist. Species-specific fly attraction was due to the differences between plant species in floral scents, floral morphology, colour patterns, and presence of other functional structures, e.g., vibratile trichomes, which were shown to contribute to pollinator attraction in one study species. The combination of these olfactory and visual cues differentially influenced pollinator preferences and thus hindered heterospecific visitation. Furthermore, a pollinator exchange experiment also highlighted that species integrity is maintained through efficient ethological isolation (pollinator attraction). The mechanical isolation mediated by the fit between floral morphology and size and/or shape of fly pollinators appears less pronounced here, but whether or not the morphological match between male (pollinium) and female (guide rails) reproductive organs can impede hybridization remains to be investigated.