Theoretical calculation on degradation mechanism of novel copolyesters under CALB enzyme.

Poly(butylene succinate-co-furandicarboxylate) (PBSF) and poly(butylene adipate-co-furandicarboxylate) (PBAF) are novel furandicarboxylic acid-based biodegradable copolyesters with great potential to replace fossil-derived terephthalic acid-based copolyesters such as poly(butylene succinate-co-terephthalate) (PBST) and poly(butylene adipate-co-terephthalate) (PBAT). In this study, quantum chemistry techniques after molecular dynamics simulations are employed to investigate the degradation mechanism of PBSF and PBAF catalyzed by Candida antarctica lipase B (CALB). Computational analysis indicates that the catalytic reaction follows a four-step mechanism resembling the ping-pong bibi mechanism, with the initial two steps being acylation reactions and the subsequent two being hydrolysis reactions. Notably, the first step of the hydrolysis is identified as the rate-determining step. Moreover, by introducing single-point mutations to expand the substrate entrance tunnel, the catalytic distance of the first acylation step decreases. Additionally, energy barrier of the rate-determining step is decreased in the PBSF system by site-directed mutations on key residues increasing hydrophobicity of the enzyme's active site. This study unprecedently show the substrate binding pocket and hydrophobicity of the enzyme's active site have the potential to be engineered to enhance the degradation of copolyesters catalyzed by CALB.

XylR regulates genes at xyl cluster, involved in D-xylose catabolism in Herbaspirillum seropedicae Z69.

D-xylose, one of the most abundant sugars in lignocellulosic biomass, is not widely used to produce bioproducts with added value, in part due to the absence of industrial microorganisms able to metabolize it efficiently. Herbaspirillum seropedicae Z69 is a ß-proteobacterium able to accumulate poly-3-hydroxybutyrate, a biodegradable thermoplastic biopolymer, with contents higher than 50%. It metabolizes D-xylose by non-phosphorylative pathways. In the genome of Z69, we found the genes xylFGH (ABC D-xylose transporter), xylB, xylD, and xylC (superior non-phosphorylative pathway), and the transcriptional regulator xylR, forming the xyl cluster. We constructed the knock-out mutant Z69ΔxylR that has a reduced growth in D-xylose and in D-glucose, compared with Z69. In addition, we analyzed the expression of xyl genes by RT-qPCR and promoter fusion. These results suggest that XylR activates the expression of genes at the xyl cluster in the presence of D-xylose. On the other hand, XylR does not regulate the expression of xylA, mhpD (lower non-phosphorylative pathways) and araB (L-arabinose dehydrogenase) genes. The participation of D-glucose in the regulation mechanism of these genes must still be elucidated. These results contribute to the development of new strains adapted to consume lignocellulosic sugars for the production of value-added bioproducts.

Characterization of Bacillus pacificus KUMBNGBT-39-Derived Polyhydroxybutyrate and Impact of Physicochemical Factors on its Production.

The extensive use of various chemicals in synthetic plastics is toxic and threatens the biosphere. To address this, the study aimed to isolate, screen, characterize, optimize, and quantify polyhydroxybutyrate (PHB)-producing bacteria using cost-effective residues. Isolated from a landfill site, the Gram-positive, rod-shaped, spore-forming, motile bacterium with intracellular PHB granules was identified as Bacillus pacificus based on phenotypic and genotypic characteristics. Optimal PHB production parameters included a nutrient broth medium, 72 h of incubation, a temperature of 37° C, a pH of 7.0, glucose as the carbon source, ammonium chloride as the nitrogen source, and a carbon-to-nitrogen ratio of 4:1, resulting in a 1.42-fold PHB production increase. B. pacificus was also cultured on various low-cost substrates. Among the oil wastes, feedstock showed the highest PHB production (1.983 ± 0.005 g/L) and among agricultural residues, the maximum PHB was obtained from rice bran (1.626 ± 0.01 g/L). UV-visible spectrophotometric, FT-IR, and HR-LCMS analysis of extracted PHB confirmed characteristics of PHB molecules (ʎ-max at 210 nm, functional groups between 1152 and 2925 cm-1). The 1H NMR analysis revealed distinct signals for protons resonating at aliphatic CH3 proton groups, bridged CH protons, and shielding CH2 proton regions that matched PHBs. Thermal gravimetric analysis (TGA) and direct scanning colorimetric (DSC) analysis revealed 89.4% degradation and melting temperature at 124.1 °C for the extracted PHB compound.

Pseudohoeflea coraliihabitans sp. nov., a poly-ß-hydroxybutyrate-producing, halotolerant bacterium isolated from coral sediment in the Dapeng peninsula (Guangdong, China).

A Gram-stain-negative, strictly aerobic, motile, flagellated, rod-shaped, halotolerant, and poly-ß-hydroxyalkanoate-producing bacterium, designated DP4N28-3T, was isolated from offshore sediment surrounding hard coral in the Dapeng peninsula (Guangdong, PR China). Growth occurred at 15-35 °C (optimal at 30 °C), pH 6.0-9.5 (optimal at 6.0-7.0), and 0.0-30.0 % NaCl concentration (w/v, optimal at 0.0-2.0 %), showing halotolerance. Phylogeny based on 16S rRNA gene sequences, five housekeeping genes, and genome sequences identified Pseudohoeflea suaedae DSM 23348T (98.1 %, 16S rRNA gene sequence similarity) as the most related species to strain DP4N28-3T. Average nucleotide identity, digital DNA-DNA hybridization, and average amino acid identity values between strain DP4N28-3T and P. suaedae DSM 23348T were all below the threshold of species demarcation. Major phenotypic differences were the flagella type and the limited sources of single carbon utilization by strain DP4N28-3T, which only included acetic acid, acetoacetic acid, d-glucuronic acid, and glucuronamide. Strain DP4N28-3T harboured the class I poly-ß-hydroxyalkanoate synthase gene (phaC) and produced poly-ß-hydroxybutyrate. The fatty acids were summed feature 8 (C18 : 1 ω6c and/or C18 : 1 ω7c, 49.4 %) and C16 : 0 (13.4 %). The major cellular polar lipids consisted of phosphatidylcholine, phosphatidylethanolamine, phosphatidylmonomethylethanolamine, phosphatidylglycerol, and sulfoquinovosyl diacylglycerol. The respiratory quinone was Q-10. The results of the phylogenetic, genomic, phenotypic, and chemotaxonomic analysis indicated that the isolated strain represents the type strain of a novel species. Based on these results, strain DP4N28-3T (=MCCC 1K05639T=KCTC 82803T) is proposed as the type strain of the novel species Pseudohoeflea coraliihabitans sp. nov.

The bifunctional impact of polylactic acid microplastics on composting processes and soil-plant systems: Dynamics of microbial communities and ecological niche competition.

Although extensive research has been conducted on the environmental impact of microplastics (MPs), their effects on microorganisms during the composting process and on the compost-soil system remain unclear. Our research investigates the microbial response to polylactic acid microplastics (PLAMPs) during aerobic composting and examines how compost enriched with PLAMPs affects plants. Our findings reveal that PLAMPs play a dual role in the composting process, influencing microorganisms differently depending on the composting phase. PLAMPs reduce the relative abundance of sensitive bacterial ASVs, specifically those belonging to Limnochordaceae and Enterobacteriaceae, during composting, while increasing the relative abundance of ASVs belonging to Steroidobacteriaceae and Bacillaceae. The impact of PLAMPs on microbial community assembly and niche width was found to be phase-dependent. In the stabilization phase (S5), the presence of PLAMPs caused a shift in the core microbial network from bacterial dominance to fungal dominance, accompanied by heightened microbial antagonism. Additionally, these intricate microbial interactions can be transferred to the soil ecosystem. Our study indicates that composting, as a method of managing PLAMPs, is also influenced by PLAMPs. This influence is transferred to the soil through the use of compost, resulting in severe oxidative stress in plants. Our research is pivotal for devising future strategies for PLAMPs management and predicting the subsequent changes in compost quality and environmental equilibrium.

Timeframe of aerobic biodegradation of bioplastics differs under standard conditions and conditions simulating technological composting with biowaste.

To determine the actual timeframe of biodegradation, bioplastics (BPs) (based on polylactic acid (PLA), starch (FS), polybutylene succinate (PBS), cellulose (Cel)) were degraded with biowaste (B), which simulates real substrate technological conditions during composting. For comparison, standard conditions (with mature compost (C)) were also applied. The 90-day aerobic tests, both with C or B, were carried out at 58 ± 2 °C. This comparison enables understanding of how BPs behave in real substrate conditions and how C and B affect the time or completeness of degradation based on oxygen consumption (OC) for BPs, the ratio of OC to theoretical oxygen consumption (OC/Th-O2), and the decrease in volatile solids (VS). Additionally, for deeper insight into the biodegradation process, microscopic, microbial (based on 16S rDNA), FTIR, and mechanical (tensile strength, elongation at break) analyses were performed. There was no association between the initial mechanical properties of BPs and the time necessary for their biodegradation. BPs lost their mechanical properties and remained visible for a shorter time when degraded with C than with B. OC for Cel, FS, PLA, and PBS biodegradation was 1143, 1654, 1748, and 1211g O2/kg, respectively, which amounted to 83, 70, 69, and 60% of the theoretical OC (Th-O2), respectively. Intensive OC took place at the same time as an intensive decrease in VS content. With C, Cel was most susceptible to biodegradation (completely biodegrading within 11 days), and PLA was least susceptible (requiring 70 days for complete biodegradation). With B, however, the time required for biodegradation was generally longer, and the differences in the time needed for complete biodegradation were smaller, ranging from 45 d (FS) to 75 d (PLA). The use of C or B had the greatest effect on Cel biodegradation (10 d vs 62 d, respectively), and the least effect on PLA (70 d vs 75 d). Specific bacterial and fungal community structures were identified as potential BP biodegraders; the communities depended on the type of BPs and the substrate conditions. In conclusion, the time needed for biodegradation of these BPs varied widely depending on the specific bioplastic and the substrate conditions; the biodegradability decreased in the following order: Cel â‰« FS â‰« PBS â‰« PLA with C and FS â‰« Cel = PBS â‰« PLA with B. The biodegradability ranking of BPs with B was assumed to be ultimate as it simulates the real substrate conditions during composting. However, all of the BPs completely biodegraded in less than 90 days.

Biodegradation of poly(butylene adipate terephthalate) and poly(vinyl alcohol) within aquatic pathway.

Understanding the environmental fate of biodegradable plastics in aquatic systems is crucial, given the alarming amount of plastic waste and microplastic particles transported through aquatic pathways. In particular, there is a need to analyze the biodegradation of commercialized biodegradable plastics upon release from wastewater treatment plants into natural aquatic systems. This study investigates the biodegradation behaviors of poly(butylene adipate terephthalate) (PBAT) and poly(vinyl alcohol) (PVA) in wastewater, freshwater, and seawater. Biodegradation of PBAT and PVA assessed through biochemical oxygen demand (BOD) experiments and microcosm tests revealed that the type of aquatic system governs the biodegradation behaviors of each plastic, with the highest biodegradation rate achieved in wastewater for both PBAT and PVA (25.6 and 32.2 % in 30 d, respectively). Plastic release pathway from wastewater into other aquatic systems simulated by sequential incubation in different microcosms suggested that PBAT exposed to wastewater and freshwater before reaching seawater was more prone to degradation than when directly exposed to seawater. On the other hand, PVA displayed comparable biodegradation rate regardless of whether it was directly exposed to seawater or had passed through other environments beforehand. Metagenome amplicon sequencing of 16S rRNA genes revealed distinct community shifts dependent on the type of plastics in changing environments along the simulated aquatic pathway. Several bacterial species putatively implicated in the biodegradation of PBAT and PVA are discussed. Our findings underscore the significant influence of pollution routes on the biodegradation of PBAT and PVA, highlighting the potential for wastewater treatment to facilitate rapid degradation compared to direct exposure to pristine aquatic environments.

The Discovery of Membrane Vesicle Biogenesis in the Polyhydroxybutyrate-non-producing Mutant Strain of Cupriavidus necator H16.

Extracellular membrane vesicles (MVs) caused by the artificial production of polyhydroxybutyrate (PHB) were previously detected in Escherichia coli. We herein observed MV biogenesis in the mutant strain (-PHB) of the natural PHB producer, Cupriavidus necator H16. This inverse relationship was revealed through comparative electron microscopic ana-lyses of wild-type and mutant strains. Based on these results, we speculate that a physiological relationship exists between MV biogenesis and PHB biosynthesis. Therefore, we propose the potential of MV biogenesis as a fermentative "stress marker" for monitoring the performance of target polymer-producing microbial platforms.

Enhancing biodegradation efficiency of PLA/PBAT-ST20 bioplastic using thermophilic bacteria co-culture system: New insight from structural characterization, enzyme activity, and metabolic pathways.

The rising utilization of PLA/PBAT-ST20 presents potential ecological risks stemming from its casual disposal and incomplete degradation. To solve this problem, this study investigated the degradation capabilities of PLA/PBAT-ST20 by a co-culture system comprising two thermophilic bacteria, Pseudomonas G1 and Kocuria G2, selected and identified from the thermophilic phase of compost. Structural characterization results revealed that the strains colonized the PLA/PBAT-ST20's surface, causing holes and cracks, with an increase in the carbonyl index (CI) and polydispersity index (PDI), indicating oxidative degradation. Enzyme activity results demonstrated that the co-culture system significantly enhanced the secretion and activity of proteases and lipases, promoting the breakdown of ester bonds. LC-QTOF-MS results showed that various intermediate products were obtained after degradation, ultimately participating in the TCA cycle (ko00020), further completely mineralized. Additionally, after 15-day compost, the co-culture system achieved a degradation rate of 72.14 ± 2.1 wt% for PBAT/PLA-ST20 films, with a decrease in the abundance of plastic fragments of all sizes, demonstrating efficient degradation of PLA/PBAT-ST20 films. This study highlights the potential of thermophilic bacteria to address plastic pollution through biodegradation and emphasizes that the co-culture system could serve as an ideal solution for the remediation of PLA/PBAT plastics.

Rapid effects of plastic pollution on coastal sediment metabolism in nature.

While extensive research has explored the effects of plastic pollution, ecosystem responses remain poorly quantified, especially in field experiments. In this study, we investigated the impact of polyester pollution, a prevalent plastic type, on coastal sediment ecosystem function. Strips of polyester netting were buried into intertidal sediments, and effects on sediment oxygen consumption and polyester additive concentrations were monitored over 72-days. Our results revealed a rapid reduction in the magnitude and variability of sediment oxygen consumption, a crucial ecosystem process, potentially attributed to the loss of the additive di(2-ethylhexyl) phthalate (DEHP) from the polyester material. DEHP concentrations declined by 89% within the first seven days of deployment. However, effects on SOC dissipated after 22 days, indicating a short-term impact and a quick recovery by the ecosystem. Our study provides critical insights into the immediate consequences of plastic pollution on ecosystem metabolism in coastal sediments, contributing to a nuanced understanding of the temporal variation of plastic pollution's multifaceted impacts. Additionally, our research sheds light on the urgent need for comprehensive mitigation strategies to preserve marine ecosystem functionality from plastic pollution impacts.

Key insights into mechanism and kinetics of biodegradation of poly (3-hydroxybutyrate)-based nanocomposite films in natural soil and river water environments.

The biodegradability of poly (3-hydroxybutyrate) (PHB)-based food packaging material PHB/5GS/0.7MgO, developed by incorporating 5 wt% grapeseed oil (GS) and 0.7 wt% MgO nanoparticles using solution casting route, was investigated in soil and river water environments. For comparison, the biodegradability of neat PHB films and PHB-based films loaded only with 5 wt% GS (PHB/5GS) was also studied. Remarkably, all PHB-based films showed 100 % weight loss in soil within 25 days. In contrast, the weight loss of PHB, PHB/5GS, and PHB/5GS/MgO films in river water was 27, 24, and 20 %, respectively, in 120 days. Gradual reduction in average molecular weight and carbonyl index, alongside an increase in crystallinity, opacity, and the number of chain scissions per unit mass, was observed for various PHB-based films during their degradation in soil and river water. Overall, this study demonstrated high degradation efficiency of PHB-based food packaging material in soil than in river water.

Optimized batch cultivation and scale-up of Bacillus thuringiensis for high-yield production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate).

Addition of statistically optimized concentration of electron acceptor, propionic acid (1.2 g/L) at different cultivation times (0 h, 14.86 h and 19 h) during batch cultivation of B. thuringiensis in mixed substrate (glucose and glycerol) featured production of 8 g/L of biomass and 3.57 g/L of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) containing 0.805 g/L of 3-hydroxyvalerate concentration. Successful scale up of batch cultivation from 7 L to a 70 L bioreactor was, thereafter, achieved using power/volume (P/V) criteria with maximum PHBV and biomass concentration of 3.57 g/L and 7.15 g/L respectively. Characterization of PHBV so produced was carried out using NMR, FTIR, DSC and TGA to elucidate its structure, thermal properties and stability to map their applications in society. These findings highlight the potential of the optimized batch cultivation and scale-up process in producing PHBV emphasizing its relevance in sustainable biopolymer production.

Ultrahigh carbon utilization in symbiotic biofilm-sludge denitrification systems using polymers as sole electron donors.

The polymer-based denitrification system is an effective nitrate removal process for treating low carbon/nitrogen wastewater. However, in polymer denitrification systems, carbon used for the denitrification reaction is weakly targeted. Improving the efficiency of carbon utilization in denitrification is important to reduce carbon wastage. In this study, a symbiotic biofilm-sludge denitrification system was constructed using polycaprolactone as electron donors. Results show that the carbon release amount in 120 days was 85.32±0.46 g, and the unit mass of polycaprolactone could remove 1.55±0.01 g NO3--N. Meaningfully, the targeted carbon utilization efficiency for denitrification could achieve 79%-85%. The quantitative results showed that the release of electron donors can be well matched to the demand for electron acceptors in the biofilm-sludge denitrification system. Overall, the symbiotic system can improve the nitrate removal efficiency and reduce the waste of carbon source.

Large-scale cultivation of Synechocystis sp. PCC6803 for the production of Poly(3-hydroxybutyrate) and its potential applications in the manufacturing of bulk and medical prototypes.

Polyhydroxyalkanoates (PHAs) are biopolymers produced by microorganisms under nutrient limiting conditions and in the presence of excess carbon source. PHAs have gained popularity as a sustainable alternative to traditional plastics. However, large scale production of PHAs is economically challenging due to the relatively high costs of organic carbon. Alternative options include using organisms capable of phototrophic or mixotrophic growth. This study aimed at the production of poly(3-hydroxybutyrate) P(3HB), a type of PHA, at pilot scale using the freshwater cyanobacterium Synechocystis sp. PCC6803. First, to identify optimal conditions for P(3HB) production from Synechocystis sp. PCC6803, different supplemental carbon source concentrations and salinity levels were tested at laboratory scale. The addition of 4 g/L acetate with no added NaCl led to P(3HB) accumulation of 10.7 % dry cell weight on the 28th day of cultivation. Although acetate additions were replicated in an outdoor 400 L serpentine photobioreactor, P(3HB) content was lower, implying uncontrolled conditions impact on biopolymer production efficiency. An optimized P(3HB) extraction methodology was developed to remove pigments, and the biopolymer was characterized and subjected to 3D printing (fused deposition modelling) to confirm its processability. This study thus successfully led to the large-scale production of P(3HB) using sustainable and environmentally friendly cyanobacterial fermentation.

Quantitative Analysis of Rhodobacter sphaeroides Storage Organelles via Cryo-Electron Tomography and Light Microscopy.

Bacterial cytoplasmic organelles are diverse and serve many varied purposes. Here, we employed Rhodobacter sphaeroides to investigate the accumulation of carbon and inorganic phosphate in the storage organelles, polyhydroxybutyrate (PHB) and polyphosphate (PP), respectively. Using cryo-electron tomography (cryo-ET), these organelles were observed to increase in size and abundance when growth was arrested by chloramphenicol treatment. The accumulation of PHB and PP was quantified from three-dimensional (3D) segmentations in cryo-tomograms and the analysis of these 3D models. The quantification of PHB using both segmentation analysis and liquid chromatography and mass spectrometry (LCMS) each demonstrated an over 10- to 20-fold accumulation of PHB. The cytoplasmic location of PHB in cells was assessed with fluorescence light microscopy using a PhaP-mNeonGreen fusion-protein construct. The subcellular location and enumeration of these organelles were correlated by comparing the cryo-ET and fluorescence microscopy data. A potential link between PHB and PP localization and possible explanations for co-localization are discussed. Finally, the study of PHB and PP granules, and their accumulation, is discussed in the context of advancing fundamental knowledge about bacterial stress response, the study of renewable sources of bioplastics, and highly energetic compounds.

Comprehensive Understanding on the Aging Process and Mechanism of Microplastics in the Sediment-Water Interface: Untangling the Role of Photoaging and Biodegradation.

Microplastics (MPs) in coastal wetlands have been of great concern, but information on the aging behavior of MPs in the sediment-water interface is still lacking. In this study, the contribution of a typical abiotic (photoaging) and biotic (biodegradation) process and the underlying aging pathway of MPs with different degradabilities (including polypropylene, polyethylene terephthalate, and polylactic acid) were studied. With a quantified relative importance of photoaging (>55%) vs biodegradation, the crucial contribution of photoaging on MP aging was highlighted. This was likely attributed to more generation of reactive oxygen species (ROS) under sunlight irradiation conditions, containing O2•- and H2O2. By raising higher the level of malondialdehyde (0.5-3.5 times as high as that in the dark condition), these photochemically formed ROS caused oxidative stress and inhibited the selective attachment of plastic-degrading microbes on the MP surface, thereby weakening the effect of biodegradation. On this basis, the aging characteristics and potential pathway of different MPs were revealed. The functional group of nondegradable polypropylene tends to be broken by ROS first, while biodegradation (Arthrobacter oryzae and Bacillus sp.) played a relatively dominant role in biodegradable polylactic acid. This study provides a new sight for the understanding on the aging behaviors of MPs in the sediment-water interface.

Insights into soil autotrophic ammonium oxidization under microplastics stress: Crossroads of nitrification, comammox, anammox and Feammox.

Microplastics (MPs) are widespread in agroecosystems and profoundly impact soil microbiome and nutrient cycling. However, the effects of MPs on soil autotrophic ammonium oxidization processes, including nitrification, complete ammonium oxidation (comammox), anaerobic ammonium oxidation (anammox), and anaerobic ammonium oxidation coupled to iron reduction (Feammox), remain unclear. These processes are the rate-limiting steps of nitrogen cycling in agroecosystems. Here, our work unveiled that exposures of polyethylene (PE), polypropylene (PP), polylactic acid (PLA), and polybutylene adipate terephthalate (PBAT) MPs significantly modulated ammonium oxidization pathways with distinct type- and dose-dependent effects. Nitrification remained the main contributor (56.4-70.7 %) to soil ammonium removal, followed by comammox (11.7-25.6 %), anammox (5.0-20.2 %) and Feammox (3.3-11.6 %). Compared with conventional nonbiodegradable MPs (i.e., PE and PP), biodegradable MPs (i.e., PLA and PBAT) exhibited more pronounced impacts on soil nutrient conditions and functional microbes, which collectively induced alterations in soil ammonium oxidation. Interestingly, low-dose PLA and PBAT remarkably enhanced the roles of anammox and Feammox in soil ammonium removal, contributing to the mitigation of soil acidification in agroecosystems. This study highlights the diverse responses of ammonium oxidization pathways to MPs, further deepening our understanding of how MPs affect biogeochemical cycling and enriching strategies for agricultural managements amid increasing MPs pollution.

Enhancing soil gross nitrogen transformation through regulation of microbial nitrogen-cycling genes by biodegradable microplastics.

Microplastics (MPs) in agricultural plastic film mulching system changes microbial functions and nutrient dynamics in soils. However, how biodegradable MPs impact the soil gross nitrogen (N) transformations and crop N uptake remain significantly unknown. In this study, we conducted a paired labeling 15N tracer experiment and microbial N-cycling gene analysis to investigate the dynamics and mechanisms of soil gross N transformation processes in soils amended with conventional (polyethylene, PE) and biodegradable (polybutylene adipate co-terephthalate, PBAT) MPs at concentrations of 0 %, 0.5 %, and 2 % (w/w). The biodegradable MPs-amended soils showed higher gross N mineralization rates (0.5-16 times) and plant N uptake rates (16-32 %) than soils without MPs (CK) and with conventional MPs. The MPs (both PE and PBAT) with high concentration (2 %) increased gross N mineralization rates compared to low concentration (0.5 %). Compare to CK, MPs decreased the soil gross nitrification rates, except for PBAT with 2 % concentration; while PE with 0.5 % concentration and PBAT with 2 % concentration increased but PBAT with 0.5 % concentration decreased the gross N immobilization rates significantly. The results indicated that there were both a concentration effect and a material effect of MPs on soil gross N transformations. Biodegradable MPs increased N-cycling gene abundance by 60-103 %; while there was no difference in the abundance of total N-cycling genes between soils without MPs and with conventional MPs. In summary, biodegradable MPs increased N cycling gene abundance by providing enriched nutrient substrates and enhancing microbial biomass, thereby promoting gross N transformation processes and maize N uptake in short-term. These findings provide insights into the potential consequences associated with the exposure of biodegradable MPs, particularly their impact on soil N cycling processes.

Characteristics of biofilms on polylactic acid microplastics and their inhibitory effects on the growth of rice seedlings: A comparative study of petroleum-based microplastics.

Increasing evidence highlights the negative effects of microplastics (MPs) on crops and bio-based plastics offer an alternative to conventional plastics. However, there is limited knowledge on the impacts and mechanisms of bio-based MPs on crop physiology. In this study, bio-based polylactic acid (PLA) and petroleum-based MPs [polyamide (PA) and polypropylene (PP)] were added to hydroponic cultures planted with rice (Oryza sativa L.) seedlings to assess their toxicity. Compared to PA and PP MPs, PLA MPs experienced greater aging after 28 days of exposure, and their surfaces were loaded with more rod-shaped microorganisms with potential plastic degradation ability, such as Proteobacteria and Bacteroidota, which competed with rice seedlings for carbon and nitrogen sources for self-multiplication, thus altering the carbon fixation and nitrogen cycling processes during rice seedling growth. Down-regulation of amino acid and lipid metabolisms in the PLA treatment inhibited the normal synthesis of chlorophyll in rice seedling leaves. Consequently, decreases in the biomass and height of rice seedling roots and shoots were observed in the PLA MP treatment. This study provides evidence that bio-based MPs may have a more severe impact on crop growth than petroleum-based MPs.

A Homolog of the Histidine Kinase RetS Controls the Synthesis of Alginates, PHB, Alkylresorcinols, and Motility in Azotobacter vinelandii.

The two-component system GacS/A and the posttranscriptional control system Rsm constitute a genetic regulation pathway in Gammaproteobacteria; in some species of Pseudomonas, this pathway is part of a multikinase network (MKN) that regulates the activity of the Rsm system. In this network, the activity of GacS is controlled by other kinases. One of the most studied MKNs is the MKN-GacS of Pseudomonas aeruginosa, where GacS is controlled by the kinases RetS and LadS; RetS decreases the kinase activity of GacS, whereas LadS stimulates the activity of the central kinase GacS. Outside of the Pseudomonas genus, the network has been studied only in Azotobacter vinelandii. In this work, we report the study of the RetS kinase of A. vinelandii; as expected, the phenotypes affected in gacS mutants, such as production of alginates, polyhydroxybutyrate, and alkylresorcinols and swimming motility, were also affected in retS mutants. Interestingly, our data indicated that RetS in A. vinelandii acts as a positive regulator of GacA activity. Consistent with this finding, mutation in retS also negatively affected the expression of small regulatory RNAs belonging to the Rsm family. We also confirmed the interaction of RetS with GacS, as well as with the phosphotransfer protein HptB.