Project description:Biodegradable plastics are one possible solution for reducing plastic waste, yet the mechanisms and organisms involved in their degradation in the aquatic environment remain understudied. In this study, we have enriched a microbial community from North Sea water and sediment, capable of growing on the polyester poly(butylene succinate). This culture was grown on two other biodegradable polyesters, polycaprolactone and ecovio® FT (a PBAT-based blended biodegradable plastic), and the differences between community structure and activity on these three polymers were determined by metagenomics and metaproteomics. We have seen that the plastic supplied drives the community structure and activity. Setups growing on ecovio® FT were more diverse, yet showed the lowest degradation, while poly(butylene succinate) and polycaprolactone resulted in a less diverse community but much higher degradation efficiencies. The dominating species were Alcanivorax sp., Thalassobius sp., or Pseudomonas sp., depending on the polymer supplied. Furthermore, we have observed that Gammaproteobacteria were more abundant and active within the biofilm and Alphaproteobacteria within the free-living fraction of the enrichments. Two of the three PETase-like enzymes isolated were expressed as tandems (Ple -tan1 &Ple – tan2) and all three were produced by Pseudomonas sp. Of those, Ple-tan1 was most active on all three substrates and also the most thermostable. Overall, we could show that all three plastics investigated can be mineralized by bacteria naturally occurring within the marine environment and characterize some of the enzymes involved in the degradation process.
Project description:Objective: To identify the pathogen causing fungemia in a Chinese patient and describe its morphological and molecular characterizes. Methods: Samples of central and peripheral venous blood were collected for blood culture. Morphology and drug sensitivities of the isolated yeast-like fungus were analyzed. rDNA sequencing and molecular phylogenetic analysis of the isolated strains were performed using DNAMAN and MEGA software. Results: A strain of yeast-like fungi was repeatedly isolated from blood samples of a Chinese patient. The isolates grew well on sabouraud medium broth plate. The colonies were smooth and round at 28°C, and were of rough surface and irregular shape at 35°C. Molecular phylogenetic trees constructed based on the internal transcribed spacer (ITS) and D1/D2 domains of 28S rDNA gene demonstrated the isolated yeast-like fungus was Moesziomyces antarcticus. Drug susceptibility test showed that this isolated M. antarcticus was resistant or had relatively low susceptibility to flucytosine, fluconazole, voriconazole, and itraconazole, and only sensitive to amphotericin. Conclusion: This study provided more information for the molecular and morphology characteristics of M. antarcticus and reviewed the species information of Moesziomyces associated with human infections, which will contribute to the identification and diagnosis of Moesziomyces infections.
Project description:The development of enzymatic processes for the environmentally friendly production of 2,5-furandicarboxylic acid (FDCA), a renewable precursor for bioplastics, from 5-hydroxymethylfurfural (HMF) has gained increasing attention over the last years. Aryl-alcohol oxidases (AAOs) catalyze the oxidation of HMF to 5-formyl-2-furancarboxylic acid (FFCA) through 2,5-diformylfuran (DFF) and have thus been applied in enzymatic reaction cascades for the production of FDCA. AAOs are flavoproteins that oxidize a broad range of benzylic and aliphatic allylic primary alcohols to the corresponding aldehydes, and in some cases further to acids, while reducing molecular oxygen to hydrogen peroxide. These promising biocatalysts can also be used for the synthesis of flavors, fragrances, and chemical building blocks, but their industrial applicability suffers from low production yield in natural and heterologous hosts. Here we report on heterologous expression of a new aryl-alcohol oxidase, MaAAO, from Moesziomyces antarcticus at high yields in the methylotrophic yeast Pichia pastoris (recently reclassified as Komagataella phaffii). Fed-batch fermentation of recombinant P. pastoris yielded around 750 mg of active enzyme per liter of culture. Purified MaAAO was highly stable at pH 2-9 and exhibited high thermal stability with almost 95% residual activity after 48 h at 57.5 °C. MaAAO accepts a broad range of benzylic primary alcohols, aliphatic allylic alcohols, and furan derivatives like HMF as substrates and some oxidation products thereof like piperonal or perillaldehyde serve as building blocks for pharmaceuticals or show health-promoting effects. Besides this, MaAAO oxidized 5-hydroxymethyl-2-furancarboxylic acid (HMFCA) to FFCA, which has not been shown for any other AAO so far. Combining MaAAO with an unspecific peroxygenase oxidizing HMFCA to FFCA in one pot resulted in complete conversion of HMF to FDCA within 144 h. MaAAO is thus a promising biocatalyst for the production of precursors for bioplastics and bioactive compounds. KEY POINTS: • MaAAO from M. antarcticus was expressed in P. pastoris at 750 mg/l. • MaAAO oxidized 5-hydroxymethyl-2-furancarboxylic acid (HMFCA). • Complete conversion of HMF to 2,5-furandicarboxylic acid by combining MaAAO and UPO.
| S-EPMC8557139 | biostudies-literature
Project description:Microbial community on biodegradable plastics
Project description:Although the biodegradation of biodegradable plastics in soil and compost is well-studied, there is little knowledge on the metabolic mechanisms of synthetic polymers degradation by marine microorganisms. Here, we present a multiomics study to elucidate the biodegradation mechanism of a commercial aromatic-aliphatic copolyester film by a marine microbial enrichment culture. The plastic film and each monomer can be used as sole carbon source. Our analysis showed that the consortium synergistically degrades the polymer, different degradation steps being performed by different members of the community. Analysis of gene expression and translation profiles revealed that the relevant degradation processes in the marine consortium are closely related to poly(ethylene terephthalate) biodegradation from terrestrial microbes. Although there are multiple genes and organisms with the potential to perform a degradation step, only a few of these are active during biodegradation. Our results elucidate the potential of marine microorganisms to mineralize biodegradable plastic polymers and describe the mechanisms of labor division within the community to get maximum energetic yield from a complex synthetic substrate.
Project description:Draft genome sequence of the yeast Pseudozyma antarctica JCM 10317 known as a producer of lipase B which used in a wide range of industrial applications