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A bacterium, Acinetobacter soli ANG344B, isolated from river water, exhibited an exceptional capacity to produce 2-phenylethanol (2-PE) using L-phenylalanine (L-Phe) as a precursor—a capability typically observed in yeasts rather than bacteria. Bioreactor experiments were conducted to evaluate the production performance, using glucose as the carbon source for cellular growth and L-Phe as the precursor for 2-PE production. Remarkably, A. soli ANG344B achieved a 2-PE concentration of 2.35 ± 0.26 g/L in just 24.5 h of cultivation, exhibiting a global volumetric productivity of 0.10 ± 0.01 g/L.h and a production yield of 0.51 ± 0.01 g2-PE/gL-Phe, a result hitherto reported only for yeasts. These findings position A. soli ANG344B as a highly promising microorganism for 2-PE production.Whole-genome sequencing of A. soli strain ANG344 revealed a genome size of 3.52 Mb with a GC content of 42.7 %. Utilizing the Rapid Annotation using Subsystem Technology (RAST) server, 3418 coding genes were predicted, including genes coding for enzymes previously associated with the metabolic pathway of 2-PE production in other microorganisms, yet unreported in Acinetobacter species. Through gene mapping, 299 subsystems were identified, exhibiting 30 % subsystem coverage. The whole genome sequence data was submitted to NCBI GeneBank with the BioProject ID PRJNA982713. These draft genome data offer significant potential for exploiting the biotechnological capabilities of A. soli strain ANG344 and for conducting further comparative genomic studies.

Members of the genus Tetrasphaera are putative polyphosphate accumulating organisms (PAOs) that have been found in greater abundance than Accumulibacter in many full-scale enhanced biological phosphorus removal (EBPR) wastewater treatment plants worldwide. Nevertheless, previous studies on the effect of environmental conditions, such as pH, on the performance of EBPR have focused mainly on the response of Accumulibacter to pH changes. This study examines the impact of pH on a Tetrasphaera PAO enriched culture, over a pH range from 6.0 to 8.0 under both anaerobic and aerobic conditions, to assess its impact on the stoichiometry and kinetics of Tetrasphaera metabolism. It was discovered that the rates of phosphorus (P) uptake and P release increased with an increase of pH within the tested range, while PHA production, glycogen consumption and substrate uptake rate were less sensitive to pH changes. The results suggest that Tetrasphaera PAOs display kinetic advantages at high pH levels, which is consistent with what has been observed previously for Accumulibacter PAOs. The results of this study show that pH has a substantial impact on the P release and uptake kinetics of PAOs, where the P release rate was >3 times higher and the P uptake rate was >2 times higher at pH 8.0 vs pH 6.0, respectively. Process operational strategies promoting both Tetrasphaera and Accumulibacter activity at high pH do not conflict with each other, but lead to a potentially synergistic impact that can benefit EBPR performance.

The composition of polyhydroxyalkanoate (PHA) monomers affects the properties and final applications of PHA polymers. This study focused on the feasibility of producing tailored PHA differing in hydroxyvalerate (HV) content, through manipulation of the acidogenic fermented stream composition, and on the characterization of PHA properties to determine the best composition for melt processing. Cheese whey was used as a feedstock, and changes in the organic loading rate during acidogenic fermentation led to the production of a fermented stream with an HV precursor content of 9–33 wt%. In the PHA production assays, a PHA content of 50 ± 11 wt%. (VSS basis) and yield of 0.76 ± 0.14 gCODPHA.gCODFP-1 were obtained. Fermented stream supplementation with HV precursors during the PHA accumulation assays indicated the feasibility of producing tailored PHA with differing HV content without a need for the selection of new cultures. The thermal properties of PHA were found to be controlled by the HV content, and PHA with approximately 30 wt% HV had the lowest melting temperature. These results demonstrated the robustness of the process at pilot scale, thus supporting full-scale applications in tailored PHA production.

Polyhydroxyalkanoates (PHAs) are natural polyesters which biodegrade in soils and oceans but have more than double the cost of comparable oil-based polymers. PHA downstreaming from its biomass represents 50% of its overall cost. Here, in an attempt to assist downstreaming, mastication of wet biomasses is tested as a new mechanical continuous biomass pretreatment with potential for industrial upscaling. Downstreaming conditions where both product recovery and purity are low due to the large amount of treated wet biomass (50% water) were targeted with the following process: extraction of 20 g in 100 mL solvent at 30 °C for 2 h, followed by 4.8 h digestion of 20 g in 0.3 M NaOH. Under the studied conditions, NaOH digestion was more effective than solvent extraction in recovering larger PHA amounts, but with less purity. A nearly 50% loss of PHA was seen during digestion after mastication. PHAs downstreamed by digestion with large amounts of impurities started to degrade at lower temperatures, but their melt elasticity was thermally stable at 170 °C. As such, these materials are attractive as fully PHA-compatible processing aids, reinforcing fillers or viscosity modifiers. On the other hand, wet biomass mastication before solvent extraction improves PHA purity and thermal stability as well as the melt rheology, which recovers the viscoelasticity measured with a PHA extracted from a dried biomass.

Bacterial cellulose is one of the most promising biomaterials for the development of a wide array of novel biotechnological solutions. Nevertheless, the commercial production of bacterial cellulose is still a challenge and obtaining novel strains presenting increased cellulose biosynthesis and stress resistance properties is of extreme importance.This work demonstrates the increased stress resistance, cellulose production abilities, and overall genomic properties of Komagataeibacter uvaceti FXV3, a novel cellulose-producing and stress resistant strain isolated from a fermented grape must.K. uvaceti FXV3 was able to grow under several stress conditions, including the presence of high concentrations of ethanol (up to 7.5 % v/v), a trait that is not observed in the model strain K. xylinus CECT 7351T. Moreover, K. uvaceti FXV3 produced increased concentrations of cellulose (4.31 mg/mL, 7 days after inoculation-DAI) when compared to K. xylinus CECT 7351T (1.42 mg/mL, 7 DAI). Moreover, the detailed analysis of strain FXV3 genome revealed the presence of several genes involved in cellulose and acetan biosynthesis, quorum-sensing and quenching mechanisms, carbohydrate, amino acid, alcohol and aldehyde metabolism, as well as several other genes involved in stress resistance. Additionally, comparative genomic analysis revealed the increased prevalence of stress resistance genes in K. uvaceti FXV3 when compared to K. xylinus CECT 7351T.Ultimately, this study reveals the increased biotechnological potential of K. uvaceti FXV3 and brings new insights into the genetics behind Komagataeibacter stress resistance and cellulose production abilities.

Background and objective: Apple pulp waste generated by the fruit processing industry is a sugar-rich material with great potential to be used as a feedstock for production of value added microbial products. The aim of this work was to use this feedstock for the cultivation of Pseudomonas citronellolis and production of medium-chain-length polyhydroxyalkanoates, a natural elastomer. Material and methods: The solid fraction of the apple pulp waste was discarded and the soluble fraction, rich in fructose (17.7 g l-1), glucose (7.5 g l-1) and sucrose (1.2 g l-1), was used for the batch bioreactor cultivation of Pseudomonas citronellolis NRRL B-2504. Results and conclusion: Pseudomonas citronellolis reached a polymer content in the biomass of 30% wt and a volumetric productivity of 0.025 g l-1 h-1. The polymer was mainly composed of 3-hydroxydecanoate (68% mol) and 3-hydroxyoctanoate (22% mol), with minor contents of 3-hydroxydodecanoate (5% mol), 3-hydroxytetradecanoate (4% mol) and 3-hydroxyhexanoate (1% mol), and had a molecular weight of 3.7×105 Da. It presented glass transition and melting temperatures of -12 and 53°C, respectively, and a thermal degradation temperature of 296°C. The polymer’s films were dense, ductile and permeable to oxygen and carbon dioxide. These results demonstrated that apple pulp waste is a suitable feedstock for the production of a biopolymer with properties that render it a promising alternative to some synthetic petrochemical-derived polyesters. Conflict of interest: The authors declare no conflict of interest.

In this research, the antibacterial and antioxidant properties of oregano essential oil (OEO), rosemary extract (RE), and green tea extract (GTE) were evaluated. These active substances were encapsulated into ultrathin fibers of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) derived from fruit waste using solution electrospinning, and the resultant electrospun mats were annealed to produce continuous films. The incorporation of the active substances resulted in PHBV films with a relatively high contact transparency, but it also induced a slightly yellow appearance and increased the films opacity. Whereas OEO significantly reduced the onset of thermal degradation of PHBV, both the RE and GTE-containing PHBV films showed a thermal stability profile that was similar to the neat PHBV film. In any case, all the active PHBV films were stable up to approximately 200 °C. The incorporation of the active substances also resulted in a significant decrease in hydrophobicity. The antimicrobial and antioxidant activity of the films were finally evaluated in both open and closed systems for up to 15 days in order to anticipate the real packaging conditions. The results showed that the electrospun OEO-containing PHBV films presented the highest antimicrobial activity against two strains of food-borne bacteria, as well as the most significant antioxidant performance, ascribed to the films high content in carvacrol and thymol. Therefore, the PHBV films developed in this study presented high antimicrobial and antioxidant properties, and they can be applied as active layers to prolong the shelf life of the foods in biopackaging applications.