Gelatin/polyvinyl alcohol films loaded with doubly stabilized clove essential oil chitosomes: Preparation, characterization, and application in packing marinated steaks.

In this study, a promising active food-packaging film of Gelatin/polyvinyl alcohol (GEL/PVA) integrated with doubly stabilized clove essential oil chitosome nanoparticles (CNP) was developed to maintain the freshness of marinated steaks. Results from the XRD and SEM experiments indicated excellent compatibility between the CNP and GEL/PVA matrix. Additionally, CNP was found to introduce more free hydroxyl groups, enhance the water retention and surface wettability of the CNP-GEL/PVA (C-G/P) film, and significantly reduce the swelling index from 963.78% to 495.11% (p < 0.05). Notably, the highest tensile strength and elongation at break (53.745 MPa and 46.536%, respectively) were achieved with the addition of 30% (v/v, based on the volume of gelatin) CNP; UVC was fully absorbed with 40% CNP; and films containing 60% CNP showed optimal inhibition of both Staphylococcus aureus and Escherichia coil, extending the shelf life of marinated steak from 3 to 7 days.

YTHDF1 facilitates esophageal cancer progression via augmenting m6A-dependent TINAGL1 translation.

N6-methyladenosine (m6A) is the most abundant internal RNA modification and plays a critical role in carcinogenesis and tumor progression. As a powerful m6A reader, YTHDF1 is implicated in multiple malignancies. However, the functions and underlying mechanisms of YTHDF1 in esophageal cancer (ESCA) are elusive. Here, we revealed that YTHDF1 expression was remarkably up-regulated in ESCA and linked with poor prognosis. Functionally, YTHDF1 promoted ESCA cell proliferation, migration, and metastasis in vitro and in vivo. Mechanistically, we demonstrated that TINAGL1 might be a potential target of YTHDF1. We revealed that YTHDF1 recognized and bound to m6A-modified sites of TINAGL1 mRNA, resulting in enhanced translation of TINAGL1. Furthermore, TINAGL1 knockdown partially rescued tumor-promoting effects of YTHDF1 overexpression. Therefore, we unveil that YTHDF1 facilitates ESCA progression by promoting TINAGL1 translation in an m6A-dependent manner, which offers an attractive therapeutic target for ESCA.

Development and characterization of wax-bovine bone protein-grapeseed oil composite oleogels: Experimental and molecular simulation studies.

Three new types of composite oleogel formulations were designed. Specifically, oleogels were prepared using 90% grapeseed oil as the oil phase and carnauba wax (CW)/beeswax/rice bran wax-bovine bone protein (BBP) as gelators. All samples were solid and had an oil-binding capacity of >90%. BBP addition considerably improved the waxy texture of the oleogel and had an important effect on the crystalline network. X-ray diffractometry indicated that BBP increased the ß'-crystal content. All samples showed sol-gel thermodynamic behavior under temperature scanning. Fourier-transform infrared spectroscopy and molecular docking confirmed the formation of noncovalent interactions dominated by van der Waals forces during the development of the oleogel. The optimal components of the three oleogels exhibited an excellent effect of slowing down the release of free fatty acids. This study could serve as a reference for the development and application of wax-protein as a new binary gelator in the food industry.

Characterization of Degraded Konjac Glucomannan from an Isolated Bacillus licheniformis Strain with Multi-Enzyme Synergetic Action.

The large molecular weight and high viscosity of natural konjac glucomannan (KGM) limit its industrial application. Microbial degradation of low-molecular-weight KGM has health benefits and various biological functions; however, the available KGM strains used in the industry have microbial contamination and low degradation efficiencies. Therefore, exploring novelly adaptable strains is critical for industrial processes. Here, the Bacillus licheniformis Z7-1 strain isolated from decaying konjac showed high efficiency for KGM degradation. The monosaccharide composition of the degradation products had a reduced molar ratio of mannose to glucose, indicating that Z7-1 preferentially degraded glucose in KGM. The degraded component was further characterized by ESI-MS, Fourier-transform infrared spectroscopy (FT-IR), and scanning electron microscopy (SEM), and it also exhibited good antibacterial activity against various food-spoilage bacteria. Genome sequencing and zymolytic analysis revealed that abundant carbohydrate-active enzymes exist in the Z7-1 genome, with at least five types of extracellular enzymes responsible for KGM degradation, manifesting multi-enzyme synergetic action. The extracellular enzymes had significant thermal stability, indicating their potential application in industry. This study provides an alternative method for obtaining low-molecular-weight KGM with antibacterial functions and supports foundational knowledge for its development as a biocatalyst for the direct conversion of biomass polysaccharides into functional components.

Structural insight of cell surface sugars in viral infection and human milk glycans as natural antiviral substance.

Viral infections are caused by the adhesion of viruses to host cell receptors, including sialylated glycans, glycosaminoglycans, and human blood group antigens (HBGAs). Atomic-level structural information on the interactions between viral particles or proteins with glycans can be determined to provide precise targets for designing antiviral drugs. Milk glycans, existing as free oligosaccharides or glycoconjugates, have attracted increasing attention; milk glycans protect infants against infectious diseases, particularly poorly manageable viral infections. Furthermore, several glycans containing structurally distinct sialic acid/fucose/sulfate modifications in human milk acting as a "receptor decoy" and serving as the natural antiviral library, could interrupt virus-receptor interaction in the first line of defense for viral infection. This review highlights the basis of virus-glycan interactions, presents specific glycan receptor binding by gastroenterovirus viruses, including norovirus, enteroviruses, and the breakthroughs in the studies on the antiviral properties of human milk glycans, and also elucidates the role of glycans in respiratory viruses infection. In addition, recent advances in methods for performing virus/viral protein-glycan interactions were reported. Finally, we discuss the prospects and challenges of the studies on the clinical application of human milk glycan for viral interventions.

Metagenomic analysis of a thermophilic bacterial consortium and its use in the bioremediation of a petroleum-contaminated soil.

Biodegradation is difficult at high temperatures due to the limited capacity of microorganisms to survive and function outside their optimum temperature range. Here, a thermophilic petroleum-degrading consortium was enriched from compost at a temperature of 55 °C. 16S rDNA and metagenomic techniques were used to analyze the composition of the consortium and the mechanisms of degradation. The consortium degraded 17000 mg total petroleum hydrocarbons (TPHs) L-1 with a degradation efficiency of 81.5% in 14 days. The consortium utilized a range of substrates such as n-hexadecane, n-docosane, naphthalene and pyrene and grew well over a wide range of pH (4-10) and salinity (0-90 g L-1). The hydrocarbon-degrading extremophilic consortium contained, inter alia, (relative abundance >1%) Caldibacillus, Geobacillus, Mycolicibacterium, Bacillus, Chelatococcus, and Aeribacillus spp. Metagenomic analysis was conducted to discover the degradation and environmental tolerance functional genes of the consortium. Two alkane hydroxylase genes, alkB and ladA, were found. A microcosm study shows that the consortium promoted the bioremediation of soil TPHs. The results indicate that the consortium may be a good candidate for the high-temperature bioremediation of petroleum-contaminated soils.

Diagnostic Value of Single LH and LH/FSH Ratio at 60-minute after GnRHa Stimulation Test for Central Precocious Puberty.

OBJECTIVES: To evaluate the diagnostic value of luteinizing hormone (LH) and LH/follicle stimulating hormone (FSH) ratio at 60 min after gonadotropin-releasing hormone analogs (GnRHa) stimulation test for central precocious puberty (CPP) in girls. METHODS: Two hundred and fifty-seven girls, aged 3 to 7.5 y, suspected of precocious puberty at authors' hospital from April 2020 through November 2023 were enrolled in the study. The blood was taken at 0, 30, 60 min after GnRHa stimulation test, and LH and LH/FSH were detected by chemiluminescence assay. The diagnostic efficacy was analysed by Mann-Whitney U test, spearman's correlation analysis and receiver operating characteristic (ROC) analysis. The proportion of obesity was analysed by Chi-square test. RESULTS: LH and LH/FSH at different times were statistically significantly different (P <0.05) between the CPP and non-CPP groups. Spearman's correlation analysis showed that the level of LH and LH/FSH at 60 min had the strongest consistency with the peak of LH (r = 0.9988, P <0.001) and LH/FSH (r = 0.9981, P <0.001). ROC curve analysis showed that the area under the ROC curves at 60 min of LH and LH/FSH were 0.975 and 0.997 with a cut-off value of 5.70 IU/L and 0.609, respectively. CONCLUSIONS: The peak of LH and LH/FSH in the diagnosis of CPP can be determined by LH and LH/FSH at 60 min after the triptorelin acetate is injected. This will reduce the number of blood draws required compared with the traditional stimulation test, which is more effective and acceptable for children.

Assessing the structural and foaming property changes in egg yolk proteins due to malondialdehyde: Experimental and molecular docking studies.

This study evaluated the effects of varying levels of malondialdehyde (MDA) on the structural and foaming properties of the egg yolk proteins (EYPs), and the interaction between them was explored by molecular docking. The results showed that oxidative modification due to MDA increased the carbonyl content of EYPs by 4.49 times. Simultaneously, the total sulfhydryl content was reduced by 21.47%, and the solubility of EYPs was significantly decreased (p < 0.05). Continuous oxidation disorders the previously ordered structure of EYPs. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated that some proteins underwent crosslinking and aggregation with increased MDA oxidation, aligning with changes in particle size and zeta-potential. Moderate oxidation (<1 mmol/L) enhanced the foaming capacity and foam stability of EYPs. Additionally, molecular docking results uncovered favorable interactions between MDA and specific EYPs, primarily through hydrogen bonding. This research offers valuable insights into managing the functional and quality changes of yolk products during processing.

Discovery, Structure, and Engineering of a cis-Geranylfarnesyl Diphosphate Synthase.

cis-Prenyltransferases (cis-PTs) catalyze the sequential head-to-tail condensation of isopentenyl diphosphate (IPP) to allylic diphosphates, producing mixed E-Z prenyl diphosphates of varying lengths; however, the specific enzymes synthesizing cis-C25 prenyl diphosphates have not been identified. Herein, we present the discovery and characterization of a cis-geranylfarnesyl diphosphate synthase (ScGFPPS) from Streptomyces clavuligerus. This enzyme demonstrates high catalytic proficiency in generating six distinct cis-polyisoprenoids, including three C25 and three C20 variants. We determined the crystal structure of ScGFPPS. Additionally, we unveil the crystal structure of nerylneryl diphosphate synthase (NNPS), known for synthesizing an all-cis-C20 polyisoprenoid. Comparative structural analysis of ScGFPPS and NNPS has identified key differences that influence product specificity. Through site-directed mutagenesis, we have identified eight single mutations that significantly refine the selectivity of ScGFPPS for cis-polyisoprenoids. Our findings not only expand the functional spectrum of cis-PTs but also provide a structural comparison strategy in cis-PTs engineering.

Discovery and biosynthesis of bacterial drimane-type sesquiterpenoids from Streptomyces clavuligerus.

Drimane-type sesquiterpenoids (DMTs) are characterized by a distinctive 6/6 bicyclic skeleton comprising the A and B rings. While DMTs are commonly found in fungi and plants, their presence in bacteria has not been reported. Moreover, the biosynthetic pathways for DMTs have been primarily elucidated in fungi, with identified P450s only acting on the B ring. In this study, we isolated and characterized three bacterial DMTs, namely 3ß-hydroxydrimenol (2), 2α-hydroxydrimenol (3), and 3-ketodrimenol (4), from Streptomyces clavuligerus. Through genome mining and heterologous expression, we identified a cav biosynthetic gene cluster responsible for the biosynthesis of DMTs 2-4, along with a P450, CavA, responsible for introducing the C-2 and C-3 hydroxy groups. Furthermore, the substrate scope of CavA revealed its ability to hydroxylate drimenol analogs. This discovery not only broadens the known chemical diversity of DMTs from bacteria, but also provides new insights into DMT biosynthesis in bacteria.

Co-regulation of Thermosensor Pathogenic Factors by C-di-GMP-Related Two-Component Systems and a cAMP Receptor-like Protein (Clp) in Stenotrophomonas maltophilia.

Stenotrophomonas maltophilia is a major threat to the food industry and human health owing to its strong protease production and biofilm formation abilities. However, information regarding regulatory factors or potential mechanisms is limited. Herein, we observed that temperature differentially regulates biofilm formation and protease production, and a cAMP receptor-like protein (Clp) negatively regulates thermosensor biofilm formation, in contrast to protease synthesis. Among four c-di-GMP-related two-component systems (TCSs), promoter fusion analysis revealed that clp transcription levels were predominantly controlled by LotS/LotR, partially controlled by both RpfC/RpfG and a novel TCS Sm0738/Sm0737, with no obvious effect caused by Sm1912/Sm1911. Biofilm formation in Δclp and ΔTCSs strains suggested that LotS/LotR controlled biofilm formation in a Clp-mediated manner, whereas both RpfC/RpfG and Sm0738/Sm0737 may occur in a distinct pathway. Furthermore, enzymatic activity analysis combined with c-di-GMP level indicated that the enzymatic activity of c-di-GMP-related metabolism proteins may not be a vital contributor to changes in c-di-GMP level, thus influencing physiological functions. Our findings elucidate that the regulatory pathway of c-di-GMP-related TCSs and Clp in controlling spoilage or the formation of potentially pathogenic factors in Stenotrophomonas expand the understanding of c-di-GMP metabolism and provide clues to control risk factors of S. maltophilia in food safety.

Diversely regio-oxidative degradation of konjac glucomannan by lytic polysaccharide monooxygenase AA10 and generating antibacterial hydrolysate.

Konjac glucomannan (KGM) hydrolysate exhibit various biological activities and health-promoting effects. Lytic polysaccharide monooxygenases (LPMOs) play an important role on enzymatic degradation of recalcitrant polysaccharides to obtain fermentable sugars. It is generally accepted that LPMOs exhibits high substrate specificity and oxidation regioselectivity. Here, a bacteria-derived SmAA10A, with chitin-active with strict C1 oxidation, was used to catalyse KGM degradation. Through ethanol precipitation, two hydrolysed KGM components (4 kDa (KGM-1) and 5 kDa (KGM-2)) were obtained that exhibited antibacterial activity against Staphylococcus aureus. In natural KGM, KGM-1, and KGM-2, the molar ratios of mannose to glucose were 1:2.19, 1:3.05, and 1:2.87, respectively, indicating that SmAA10A preferentially degrades mannose in KGM. Fourier-transform infrared spectroscopy and scanning electron microscopy imaging revealed the breakage of glycosylic bonds during enzymatic catalysis. The regioselectivity of SmAA10A for KGM degradation was determined based on the fragmentation behaviour of the KGM-1 and KGM-2 oligosaccharides and their NaBD4-reduced forms. SmAA10A exhibited diverse oxidation degradation of KGM and generated single C1-, single C4-, and C1/C4-double oxidised oligosaccharide forms. This study provides an alternative method for obtaining KGM degradation components with antibacterial functions and expands the substrate specificity and oxidation regioselectivity of bacterial LPMOs.

Oxidative Modification, Structural Conformation, and Gel Properties of Pork Paste Protein Mediated by Oxygen Concentration in Modified Atmosphere Packaging.

The objective of this study was to investigate the effect of pork oxidation through modified atmosphere packaging (MAP) on gel characteristics of myofibrillar proteins (MP) during the heat-induced gelation process. The pork longissimus thoracis (LT) was treated by MAP at varying oxygen concentrations (0, 20, 40, 60, and 80% O2) with a 5-day storage at 4 °C for the detection of MP oxidation and gel properties. The findings showed the rise of O2 concentration resulted in a significant increase of carbonyl content, disulfide bond, and particle size, and a decrease of sulfhydryl content and MP solubility (p < 0.05). The gel textural properties and water retention ability were significantly improved in MAP treatments of 0-60% O2 (p < 0.05), but deteriorated at 80% O2 level. As the concentration of O2 increased, there was a marked decrease in the α-helix content within the gel, accompanied by a simultaneous increase in ß-sheet content (p < 0.05). Additionally, a judicious oxidation treatment (60% O2 in MAP) proved beneficial for crafting dense and uniform gel networks. Our data suggest that the oxidation treatment of pork mediated by O2 concentration in MAP is capable of reinforcing protein hydrophobic interaction and disulfide bond formation, thus contributing to the construction of superior gel structures and properties.

Construction of Hospice Care Evaluation System for Terminally Ill Patients in ICU.

Background: Terminally ill patients can benefit from hospice care, which specifically addresses the needs of patients and families affected by terminal illness. However, there is a lack of standardized evaluation criteria to assess the quality of hospice care for terminally ill patients in the ICU, and it is impossible to evaluate the service quality of hospice care. To use the Delphi method to construct a hospice care system for terminally ill patients in ICU that meets clinical needs, and to provide theoretical support for nursing decision-making of terminally ill patients in clinical ICU. Methods: Obtain relevant literatures by entering specific key words into the database, the hospice care nursing system for terminally ill patients in ICU was preliminarily drawn up by literature analysis, and 24 experts in this field were consulted for 3 rounds by Delphi method to discuss the development status of hospice care and finally establish the hospice care nursing system. Results: In the three rounds of letter inquiries, the positive coefficients of experts were all high, the expert authority coefficient (Cr) were 0.864, 0.849, 0.832, and the expert opinion coordination coefficient(W) were 0.186, 0.319, 0.224; The system includes 8 first-level indicators, 27 second-level indicators and 9 third-level indicators. Conclusion: In this study, three rounds of Delphi consultation methods were used to construct an evaluation index system for the nursing quality of hospice care for ICU patients. The evaluation indicators formulated closely focus on the physiological and psychological characteristics of ICU patients, which can provide a better reference for ICU patients with advanced life in the future.

Improving the solubility and interfacial absorption of hempseed protein via a novel high pressure homogenization-assisted pH-shift strategy.

A pH shift treatment aided by high pressure homogenization (HPH) with various pressures (0-120 MPa) was employed to structurally modify hempseed protein isolate (HPI). Compared with individual pH shift or HPH treatment, HPH-assisted pH shift improved the structural flexibility of HPI, as revealed by the increased random coil in protein secondary structure. With the incorporation of HPH into pH shift, the intrinsic fluorescence intensity was remarkably attenuated and redshifted, whereas the surface hydrophobicity was pronouncedly boosted, indicating the extensive unfolding of protein structure. Moreover, the cotreated HPI exhibited a smaller and more homogenous particle size, notably at a higher pressure. Consequently, the solubility was drastically raised by the cooperated treatments, to the maximum value (62.8 %) at 120 MPa. These physicochemical changes in the cotreated HPI facilitated a consolidated interfacial activity. Moreover, the cooperated treatment, especially highly pressured (120 MPa), facilitated the penetration and rearrangement of proteins at the oil-water interface.

Modulating in vitro digestion of whey protein cold-set emulsion gels via gel properties modification with gallic acid and EGCG.

Gallic acid (GA) and epigallocatechin gallate (EGCG), cooperated at varied ratios (1:0, 3:1, 1:1, 1:3, and 0:1), were employed to modify gel properties of calcium induced-whey protein emulsion gel. The effects of GA/EGCG on emulsion morphology, as well as gel properties and in vitro digestive behavior of the emulsion gels were investigated. Compared with emulsions without phenolics, GA/EGCG induced slightly smaller particle size and stronger electrostatic repulsion between emulsion droplets. Moreover, GA/EGCG, notably at a ratio of 3:1, promoted electrostatic and hydrophobic interactions between protein molecules and the formation of a compact and filamentous gel microstructure, resulting in a remarkable increment in the gel strength (up to 106 %). Furthermore, in vitro oral digestion, dynamic gastric digestion (using an artificial gastric digestive system, AGDS), and intestinal digestion of the emulsion gels were simulated. Particle size and protein hydrolysis results revealed that GA/EGCG was prone to weaken the physical disintegration of gels, reduce protein hydrolysis, and enhance the stability of emulsified oil droplets during dynamic gastric digestion. As a consequence, delayed release of oil droplets was observed in the gels and more free fatty acids were released in the intestinal digestion, particularly in the gel with GA/EGCG (3:1). These findings would provide novel strategies for application of phenolic compounds in developing protein gel-based delivery systems.

Arabic gum grafted with phenolic acid as a novel functional stabilizer for improving the oxidation stability of oil-in-water emulsion.

Three kinds of phenolic acids: ferulic acid (FA), caffeic acid (CA), and gallic acid (GA) with different chemical structures were individually grafted onto Arabic gum (AG) via a laccase mediated method, and their roles in stabilizing o/w emulsions were evaluated. The total phenolic content in modified AG increased from 2.7 ± 0.2 to 18.7 ± 0.2, 19.8 ± 0.6, 22.4 ± 0.8 mg/g after 4 h of laccase catalysis, respectively. FTIR spectra of modified AGs exhibited additional phenolic characteristics, revealing the successful grafting of phenolic acids to AG structure. Compared with natural AG, modified AGs showed remarkably enhanced thermal stability, as well as antioxidant capacity in an order of gallic acid > caffeic acid > ferulic acid. The incorporation of phenolic acids into AG dramatically improved its emulsification performance. Herein, gallic acid-modified AG evinced up to 17.6 % and 12.6 % increments in emulsifying activity and emulsion stability relative to natural AG, respectively. Moreover, the oxidative stability of AG emulsions was pronouncedly meliorated by the introduced phenolic acids, especially gallic acid, as manifested by the suppressed production of primary and secondary oxidation products.

Comparative Analysis of Human Milk Glycosphingolipids from Different Secretor Mothers Using HILIC-MS/MS.

Glycosphingolipids participate in brain development, intestinal tract maturation, and defense against gut pathogens. Here, we performed a qualitative and quantitative comparison of milk glycosphingolipids from secretors and nonsecretors. Hydrophilic interaction chromatography-electrospray ionization-tandem mass spectrometry was employed, along with an internal standard, to resolve the complications presented by the fact that glycosphingolipids are structurally diverse, varying in glycan composition and ceramide. In total, 101 glycosphingolipids were detected, of which 76 were reported for the first time, including fucose-modified neutral glycosphingolipids. Seventy-eight glycosphingolipids differed significantly between secretor and nonsecretor milk (p < 0.05), resulting in higher levels of certain neutral species (p < 0.001) but lower levels of fucose-modified monosialylated and disialylated species in secretor mothers (p < 0.01). In both milk types, the most abundant glycosphingolipids were of the monosialylated type, followed by disialylated, neutral, and trisialylated ones. Notably, fucose-modified monosialylated glycosphingolipids accounted for the highest proportion.

Synthetic exploration of electrophilic xanthylation via powerful N-xanthylphthalimides.

Organic xanthates are broadly applied as synthetic intermediates and bioactive molecules in synthetic chemistry. Electrophilic xanthylation represents a promising approach but has rarely been explored mainly due to the lack of powerful electrophilic reagents. Herein, synthetic exploration of electrophilic xanthylation via powerful N-xanthylphthalimides was investigated. This strategy might provide a new avenue to less-concerned but meaningful electrophilic xanthylation in organic synthesis. With the help of these powerful reagents, electrophilic xanthylation of a wide range of substrates including aryl/alkenyl boronic acids, ß-keto esters, 2-oxindole, and alkyl amines, as well as previously inaccessible phenols (first report) was achieved under mild reaction conditions. Notably, this simple electrophilic xanthylation of alkyl amine substrates will occur in the desulfuration reaction, consistent with the previously reported methods. Similarly, xanthamide and thioxanthate groups could also be transformed into desired nucleophiles via this electrophilic reagent strategy. The broad substrate scope, excellent functional group compatibility and late-stage functionalization of bioactive or functional molecules made them very attractive as general reagents which will allow rapid incorporation of SC(S)R (R = OEt, Oalkyl, NEt2 and SEt) into the target molecules.

Metagenomics reveals mechanism of pyrene degradation by an enriched bacterial consortium from a coking site contaminated with PAHs.

A bacterial consortium, termed WPB, was obtained from polycyclic aromatic hydrocarbons (PAHs) contaminated soil from a coking site. The consortium effectively degraded 100 mg L-1 pyrene by 94.8 % within 12 days. WPB was also able to degrade phenanthrene (98.3 %) and benzo[a]pyrene (24.6 %) in 12 days, while the individual isolates showed no PAHs degrading ability. Paracoccus sp. dominated the bacterial consortium (65.0-86.2 %) throughout the degradation process. Metagenomic sequencing reveals the proportion of sequences with xenobiotics biodegradation and metabolism increased throughout the degradation process indicating the great potential of WPB to degrade pollutants. The annotation of genes by metagenomic analysis help reconstruct the degradation pathways ("phthalate pathway" and "naphthalene degradation") and reveal how different bacteria contribute to the degradation process. Mycobacterium gilvum was found to carry nidAB genes that catalyze the first step of high-molecular-weight (HMW) PAHs in the degradation process despite Mycobacterium gilvum accounting for only 0.005-0.06 %. In addition, genomes of Paracoccus denitrificans and some other genera affiliated with Devosia, Pusillimonas caeni and Eoetvoesia caeni were successfully recovered and were found to carry genes responsible for the degradation of the intermediates of pyrene. These results enable further understanding of the metabolic patterns of pyrene-degrading consortia and provide direction for further cultivation and discovery of key players in complex microbial consortia.