Ten samples of tropical lichens collected from Doi Inthanon, Thailand, were explored for the diversity of their bacterial microbiomes through 16S rRNA-based metagenomics analysis. The five predominant lichen-associated bacteria belonged to the phyla Proteobacteria (31.84%), Planctomycetota (17.08%), Actinobacteriota (15.37%), Verrucomicrobiota (12.17%), and Acidobacteriota (7.87%). The diversity analysis metric showed that Heterodermia contained the highest bacterial species richness. Within the lichens, Ramalina conduplicans and Cladonia rappii showed a distinct bacterial community from the other lichen species. The community of lichen-associated actinobacteria was investigated as a potential source of synthesized biologically active compounds. From the total Operational Taxonomic Units (OTUs) found across the ten different lichen samples, 13.21% were identified as actinobacteria, including the rare actinobacterial genera that are not commonly found, such as Pseudonocardia, Kineosporia, Dactylosporangium, Amycolatopsis, Actinoplanes, and Streptosporangium. Evaluation of the pretreatment method (heat, air-drying, phenol, and flooding) and isolation media used for the culture-dependent actinobacterial isolation revealed that the different pretreatments combined with different isolation media were effective in obtaining several species of actinobacteria. However, metagenomics analyses revealed that there were still several strains, including rare actinobacterial species, that were not isolated. This research strongly suggests that lichens appear to be a promising source for obtaining actinobacteria.
Oil palm empty fruit bunch (OPEFB) was pretreated by NaOH-steam explosion and then fermented to ethanol by separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF) processes using Kluyveromyces marxianus G2-16-1 at 40 ºC. The maximum ethanol production by the SHF and SSF processes was 8.09 g/L (22.21 g/L reducing sugar, 0.08 g/g OPEFB) and 13.658 g/L (0.136 g/g OPEFB), respectively, at 48 h. The OPEFB hydrolysate mixed with molasses to 22% (w/v, total sugar) gave an ethanol yield of 61.60 g/L (0.38 g/g total sugar) at 72 h, while molasses alone gave 53.89 g/L (0.34 g/g total sugar). The OPEFB slurry (OPEFBS; OPEFB hydrolysate containing the solid residue of pretreated OPEFB) gave a maximum ethanol yield of 68.77 g/L (0.44 g/g total sugar) when it was mixed with molasses. Scanning electron micrographs of the solid OPEFB residue in the OPEFBS showed yeast cells adsorbed to the OPEFB fibers. The results indicated that ethanol production from molasses mixed with OPEFB hydrolysate was equal to the cumulative sum of ethanol production from each raw material, and the solid OPEFB residue in the OPEFBS increased the ethanol production in the co-fermentation of molasses and OPEFB hydrolysate.
NaOH-impregnation with catalyst steam explosion was found to be an efficient pretreatment method for oil palm empty fruit bunch (OPEFB) as a substrate for oil production by Naganishia cerealis IN1S2.5. Cellulase hydrolysis of the pretreated OPEFB yielded glucose at 0.364 g/g. Investigation of N. cerealis IN1S2.5 oil production in the OPEFB hydrolysate revealed a maximum oil yield (2.46 g/L) when the C/P molar ratio of the OPEFB hydrolysate was adjusted to 25.71, supplemented with Ca2+ and Zn2+, and set to pH 4. The N. cerealis IN1S2.5 oil was comprised of oleic (37.6%), palmitic (36.2%), and steric (17.9%) acids, all (w/w), as the major fatty acids. Predicted properties of the produced biodiesel indicated the potential of N. cerealis IN1S2.5 oil as a biodiesel feedstock.
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