Phenylacetic acid, an anti-vaginitis metabolite produced by the vaginal symbiotic bacterium Chryseobacterium gleum.

The human microbiome contains genetic information that regulates metabolic processes in response to host health and disease. While acidic vaginal pH is maintained in normal conditions, the pH level increases in infectious vaginitis. We propose that this change in the vaginal environment triggers the biosynthesis of anti-vaginitis metabolites. Gene expression levels of Chryseobacterium gleum, a vaginal symbiotic bacterium, were found to be affected by pH changes. The distinctive difference in the metabolic profiles between two C. gleum cultures incubated under acidic and neutral pH conditions was suggested to be an anti-vaginitis molecule, which was identified as phenylacetic acid (PAA) by spectroscopic data analysis. The antimicrobial activity of PAA was evaluated in vitro, showing greater toxicity toward Gardnerella vaginalis and Candida albicans, two major vaginal pathogens, relative to commensal Lactobacillus spp. The activation of myeloperoxidase, prostaglandin E2, and nuclear factor-κB, and the expression of cyclooxygenase-2 were reduced by an intravaginal administration of PAA in the vaginitis mouse model. In addition, PAA displayed the downregulation of mast cell activation. Therefore, PAA was suggested to be a messenger molecule that mediates interactions between the human microbiome and vaginal health.

Genome features of a novel hydrocarbonoclastic Chryseobacterium oranimense strain and its comparison to bacterial oil-degraders and to other C. oranimense strains.

For the first time, we report the whole genome sequence of a hydrocarbonoclastic Chryseobacterium oranimense strain isolated from Trinidad and Tobago (COTT) and its genes involved in the biotransformation of hydrocarbons and xenobiotics through functional annotation. The assembly consisted of 11 contigs with 2,794 predicted protein-coding genes which included a diverse group of gene families involved in aliphatic and polycyclic hydrocarbon degradation. Comparative genomic analyses with 18 crude-oil degrading bacteria in addition to two C. oranimense strains not associated with oil were carried out. The data revealed important differences in terms of annotated genes involved in the hydrocarbon degradation process that may explain the molecular mechanisms of hydrocarbon and xenobiotic biotransformation. Notably, many gene families were expanded to explain COTT's competitive ability to manage habitat-specific stressors. Gene-based evidence of the metabolic potential of COTT supports the application of indigenous microbes for the remediation of polluted terrestrial environments and provides a genomic resource for improving our understanding of how to optimize these characteristics for more effective bioremediation.

Cellular Response and Molecular Mechanism of Glyphosate Degradation by Chryseobacterium sp. Y16C.

Glyphosate is one of the most widely used herbicides worldwide. Unfortunately, the continuous use of glyphosate has resulted in serious environmental contamination and raised public concern about its impact on human health. In our previous study, Chryseobacterium sp. Y16C was isolated and characterized as an efficient degrader that can completely degrade glyphosate. However, the biochemical and molecular mechanisms underlying its glyphosate biodegradation ability remain unclear. In this study, the physiological response of Y16C to glyphosate stimulation was characterized at the cellular level. The results indicated that, in the process of glyphosate degradation, Y16C induced a series of physiological responses in the membrane potential, reactive oxygen species levels, and apoptosis. The antioxidant system of Y16C was activated to alleviate the oxidative damage caused by glyphosate. Furthermore, a novel gene, goW, was expressed in response to glyphosate. The gene product, GOW, is an enzyme that catalyzes glyphosate degradation, with putative structural similarities to glycine oxidase. GOW encodes 508 amino acids, with an isoelectric point of 5.33 and a molecular weight of 57.2 kDa, which indicates that it is a glycine oxidase. GOW displays maximum enzyme activity at 30 °C and pH 7.0. Additionally, most of the metal ions exhibited little influence on the enzyme activity except for Cu2+. Finally, with glyphosate as the substrate, the catalytic efficiency of GOW was higher than that of glycine, although opposite results were observed for the affinity. Taken together, the current study provides new insights to deeply understand and reveal the mechanisms of glyphosate degradation in bacteria.

Genomics assisted characterization of plant growth-promoting and metabolite producing psychrotolerant Himalayan Chryseobacterium cucumeris PCH239.

Here, we report the first complete genome of a psychrotolerant and yellow-pigmented rhizobacteria Chryseobacterium cucumeris PCH239. It was obtained from the rhizospheric soil of the Himalayan plant Bergenia ciliata. The genome consists of a single contig (5.098 Mb), 36.3% G + C content, and 4899 genes. The cold adaptation, stress response, and DNA repair genes promote survivability in a high-altitude environment. PCH239 grows in temperature (10-37 °C), pH (6.0-8.0), and NaCl (2.0%). The genome derived plant growth-promoting activities of siderophore production (siderophore units 53 ± 0.6), phosphate metabolism (PSI 5.0 ± 0.8), protease, indole acetic acid production (17.3 ± 0.5 µg/ml), and ammonia (2.89 ± 0.4 µmoles) were experimentally validated. Interestingly, PCH239 treatment of Arabidopsis seeds significantly enhances germination, primary, and hairy root growth. In contrast, Vigna radiata and Cicer arietinum seeds had healthy radicle and plumule elongation, suggesting varied plant growth-promotion effects. Our findings suggested the potential of PCH239 as a bio-fertilizer and biocontrol agent in the challenging conditions of cold and hilly regions.

Enhanced protein glutaminase production from Chryseobacterium proteolyticum combining physico-chemical mutagenesis and resistance screening and its application to soybean protein isolates.

BACKGROUND: Protein glutaminase (PG) is a novel protein modification biotechnology that is increasingly being used in the food industry. However, the current level of fermentation of PG-producing strains still does not meet the requirements of industrial production. To obtain the mutant strains with high PG production, the atmospheric and room temperature plasma (ARTP) combined with LiCl chemical mutagen were used in mutagenesis of a PG producing Chryseobacterium proteolyticum 1003. RESULTS: A mutant strain (WG15) was successfully obtained based on malonic acid resistance screening after compound mutagenesis of the starting strain C. proteolyticum 1003 using ARTP with LiCl, and it was confirmed to be genetically stable in PG synthesis after 15 generations. The protein glutaminase production of WG15 was 2.91 U mL-1 after optimization of fermentation conditions, which is 48.69% higher than the original strain C. proteolyticum 1003. The PG obtained from fermentation showed good activities in deamidation of soy protein isolate. The solubility and foaming properties of the PG-treated soy protein isolate were significantly increased by 36.50% and 10.03%, respectively, when PG was added at the amount of 100 U mL-1 . In addition, the emulsifying activity and emulsion stability of the treated soy protein isolate were improved by 12.44% and 10.34%, respectively, on the addition of 10 U mL-1 PG. The secondary structure of the soy protein isolate changed after PG treatment, with an increased proportion of glutamate. CONCLUSION: The results of the present study indicate that the PG produced by this mutant strain could improve the functional properties of soybean protein isolate and the C. proteolyticum mutant WG15 has great potential in food industry. © 2023 Society of Chemical Industry.

A novel binary pesticidal protein from Chryseobacterium arthrosphaerae controls western corn rootworm by a different mode of action to existing commercial pesticidal proteins.

The western corn rootworm (WCR) Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae) remains one of the economically most important pests of maize (Zea mays) due to its adaptive capabilities to pest management options. This includes the ability to develop resistance to some of the commercial pesticidal proteins originating from different strains of Bacillus thuringiensis. Although urgently needed, the discovery of new, environmentally safe agents with new modes of action is a challenge. In this study we report the discovery of a new family of binary pesticidal proteins isolated from several Chryseobacterium species. These novel binary proteins, referred to as GDI0005A and GDI0006A, produced as recombinant proteins, prevent growth and increase mortality of WCR larvae, as does the bacteria. These effects were found both in susceptible and resistant WCR colonies to Cry3Bb1 and Cry34Ab1/Cry35Ab1 (reassigned Gpp34Ab1/Tpp35Ab1). This suggests GDI0005A and GDI0006A may not share the same binding sites as those commercially deployed proteins and thereby possess a new mode of action. This paves the way towards the development of novel biological or biotechnological management solutions urgently needed against rootworms.

Characterization of a novel glyphosate-degrading bacterial species, Chryseobacterium sp. Y16C, and evaluation of its effects on microbial communities in glyphosate-contaminated soil.

Widespread use of the herbicide glyphosate in agriculture has resulted in serious environmental problems. Thus, environment-friendly technological solutions are urgently needed for the removal of residual glyphosate from soil. Here, we successfully isolated a novel bacterial strain, Chryseobacterium sp. Y16C, which efficiently degrades glyphosate and its main metabolite aminomethylphosphonic acid (AMPA). Strain Y16C was found to completely degrade glyphosate at 400 mg·L-1 concentration within four days. Kinetics analysis indicated that glyphosate biodegradation was concentration-dependent, with a maximum specific degradation rate, half-saturation constant, and inhibition constant of 0.91459 d-1, 15.79796 mg·L-1, and 290.28133 mg·L-1, respectively. AMPA was identified as the major degradation product of glyphosate degradation, suggesting that glyphosate was first degraded via cleavage of its C-N bond prior to subsequent metabolic degradation. Strain Y16C was also found to tolerate and degrade AMPA at concentrations up to 800 mg·L-1. Moreover, strain Y16C accelerated glyphosate degradation in soil indirectly by inducing a slight alteration in the diversity and composition of soil microbial community. Taken together, our results suggest that strain Y16C may be a potential microbial agent for bioremediation of glyphosate-contaminated soil.

Nitrogen fertilization modulates beneficial rhizosphere interactions through signaling effect of nitric oxide.

Chemical nitrogen (N) fertilization is customary for increasing N inputs in agroecosystems. The nutritional effects of N fertilization on plants and soil microbes have been well studied. However, the signaling effects of N fertilization on rhizosphere plant-microbe interactions and the following feedback to plant performance remain unknown. Here, we investigated the effect of different N fertilizations on the behavior of the plant growth-promoting rhizobacteria (PGPR) Bacillus velezensis SQR9 in the cucumber (Cucumis sativus L.) rhizosphere. Moderate N fertilization promoted higher rhizosphere colonization of strain SQR9 than insufficient or excessive N input. Nitric oxide (NO) produced through the denitrification process under N fertilization was identified as the signaling molecule that dominates the root colonization of PGPR, and this effect could be neutralized by the NO-specific scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxy-3-oxide. Gene expression analysis demonstrated that NO regulated the biofilm formation of strain SQR9 by affecting the synthesis of extracellular matrix γ-polyglutamic acid, consequently impacting its root colonization. Finally, we demonstrated that moderate N fertilization-modulated enhanced PGPR root colonization can significantly promote plant growth and nitrogen use efficiency. This study provides insights into our understanding of the beneficial rhizosphere plant-microbe interactions under N fertilization and suggests that rational fertilization is critical to promote beneficial rhizosphere interactions for sustainable agricultural production.

Microbial bioconversion of feathers into antioxidant peptides and pigments and their liposome encapsulation.

OBJECTIVES: The co-encapsulation of bioactive peptides obtained from degradation of chicken feathers and flexirubin-type pigment produced by Chryseobacterium sp. kr6 into phosphatidylcholine liposomes was investigated. RESULTS: Control empty liposomes showed mean diameter of 168.5 nm, varying to 185.4, 102.0 and 98.5 nm after the encapsulation of peptides, pigment and their co-encapsulation, respectively. Control liposomes presented zeta potential of - 20.9 mV, while the formulations containing the bioactive compounds showed values of - 30 mV or higher in magnitude. Infrared analysis revealed typical spectra for phosphatidylcholine, suggesting that no new chemical bonds were formed after encapsulation. ABTS radical scavenging assay showed that the antioxidant activity of the compounds was maintained after encapsulation. CONCLUSIONS: Feather waste can be a valuable substrate for simultaneous production of antioxidant peptides and pigment by Chryseobacterium sp. kr6, and their encapsulation into liposomes may be a suitable alternative for delivery of these natural antioxidants.

Manganese-contaminated groundwater treatment by novel bacterial isolates: kinetic study and mechanism analysis using synchrotron-based techniques.

The occurrence of manganese in groundwater causes coloured water and pipe rusting in water treatment systems. Consumption of manganese-contaminated water promotes neurotoxicity in humans and animals. Manganese-oxidizing bacteria were isolated from contaminated areas in Thailand for removing manganese from water. The selected bacterium was investigated for its removal kinetics and mechanism using synchrotron-based techniques. Among 21 isolates, Streptomyces violarus strain SBP1 (SBP1) was the best manganese-oxidizing bacterium. At a manganese concentration of 1 mg L-1, SBP1 achieved up to 46% removal. The isolate also successfully removed other metal and metalloid, such as iron (81%) and arsenic (38%). The manganese concentration played a role in manganese removal and bacterial growth. The observed self-substrate inhibition best fit with the Aiba model. Kinetic parameters estimated from the model, including a specific growth rate, half-velocity constant, and inhibitory constant, were 0.095 h-1, 0.453 mg L-1, and 37.975 mg L-1, respectively. The synchrotron-based techniques indicated that SBP1 removed manganese via combination of bio-oxidation (80%) and adsorption (20%). The study is the first report on biological manganese removal mechanism using synchrotron-based techniques. SBP1 effectively removed manganese under board range of manganese concentrations. This result showed the potential use of the isolate for treating manganese-contaminated water.

Genomic and functional insights into the adaptation and survival of Chryseobacterium sp. strain PMSZPI in uranium enriched environment.

Metal enriched areas represent important and dynamic microbiological ecosystems. In this study, the draft genome of a uranium (U) tolerant bacterium, Chryseobacterium sp. strain PMSZPI, isolated from the subsurface soil of Domiasiat uranium ore deposit in Northeast India, was analyzed. The strain revealed a genome size of 3.8 Mb comprising of 3346 predicted protein-coding genes. The analysis indicated high abundance of genes associated with metal resistance and efflux, transporters, phosphatases, antibiotic resistance, polysaccharide synthesis, motility, protein secretion systems, oxidoreductases and DNA repair. Comparative genomics with other closely related Chryseobacterium strains led to the identification of unique inventory of genes which were of adaptive significance in PMSZPI. Consistent with the genome analysis, PMSZPI showed superior tolerance to uranium and other heavy metals. The metal exposed cells exhibited transcriptional induction of metal translocating PIB ATPases suggestive of their involvement in metal resistance. Efficient U binding (~90% of 100 µM U) and U bioprecipitation (~93-94% of 1 mM U at pH 5, 7 and 9) could be attributed as uranium tolerance strategies in PMSZPI. The strain demonstrated resistance to a large number of antibiotics which was in agreement with in silico prediction. Reduced gliding motility in the presence of cadmium and uranium, enhanced biofilm formation on uranium exposure and tolerance to 1.5 kGy of 60Co gamma radiation were perceived as adaptive responses in PMSZPI. Overall, the positive correlation observed between uranium/metal tolerance abilities predicted using genome analysis and the functional characterization reinforced the multifaceted adaptation strategies employed by PMSZPI for its survival in the soil of uranium ore deposit comprising of high concentrations of uranium and other heavy metals.

Microbial degradation of poultry feather biomass in a constructed bioreactor and application of hydrolysate as bioenhancer to vegetable crops.

Bioconversion of recalcitrant keratinous biomass is one of the greatest ways to utilize products of feather hydrolysis and recycle them into bionetwork. Present study revealed 87% degradation of poultry feathers within 48 h in a constructed bioreactor using Chryseobacterium sp. RBT. The resulting feather hydrolysate (FH) was rich in soluble protein (3.56 ± 0.18 mg/ml), amino acids (3.83 ± 0.20 mg/ml), and macro and micro nutrients like N (8.0302%), P (0.3876%), K (0.5532%), Cu (0.0684%), Mg (0.8078%), Mn (0.2001%), Ca (0.4832%), Zn (0.0442%), and Fe (0.0330%). HPTLC analysis of FH revealed presence of tryptophan, cysteine, methionine, phenylalanine, glycine, valine, tyrosine, lysine, leucine, and serine as the primary amino acids. Field studies were conducted to apply FH as the bioenhancer to commercially important crops like brinjal and chilli through root drenching (20%, v/v). FH showed positive impact on the growth and development of plants along with early flowering and improved crop yield. In addition, nutritional quality of brinjal and chilli in terms of protein, amino acids, reducing sugars, phenolics, flavonoids, and antioxidant was elevated. Therefore, promotion and utility of by-products generated in feather degradation would be an effective strategy focusing on sustainable agricultural practices and problems associated with the waste management.

Microbial antimonate reduction with a solid-state electrode as the sole electron donor: A novel approach for antimony bioremediation.

The anaerobic antimonate [Sb(V)] reduction with a solid-state electrode serving as the sole electron donor was demonstrated by employing a bioelectrochemical system. The highest Sb(V) reduction efficiency was observed at the biocathode potential of -0.7 V versus standard hydrogen electrode using a cathode potential range from -0.5 V to -1.1 V. The scanning electron microscopy and energy dispersive X-ray spectroscopy indicated that both amorphous and crystallized Sb2O3 were formed as products of Sb(V) reduction. The irreversible recovery of bioelectrochemical Sb(V), when the cathode potential deviated from the optimal potential, was explained through the alteration in microbial communities, which was further elucidated by the next-generation sequencing of 16S rRNA gene amplicons. Chryseobacterium koreense and Stenotrophomonas nitritireducens were the dominant species of microbial consortia at Sb(V)-reducing biocathodes. This study revealed a novel option for bioremediation of Sb at underground contaminated sites, where the delivery of organic electron donors is limited or ineffective.

A new microbial gluten-degrading prolyl endopeptidase: Potential application in celiac disease to reduce gluten immunogenic peptides.

Gluten is a complex of proteins present in barley, wheat, rye and several varieties of oats that triggers celiac disease in genetically predisposed subjects. Gluten is notoriously difficult to digest by mammalian proteolytic enzymes and therefore, proline-rich digestion-resistant peptides contain multiple immunogenic epitopes. Prolyl endopeptidases (PEP) hydrolyse internal proline residues on the carboxyl side of peptides and have been proposed for food gluten detoxification and as oral enzyme supplementation for celiacs. The aim of this study was to identify new gluten-degrading microbial enzymes with the potential to reduce gluten immunogenicity by neutralizing its antigenic epitopes. Using a gluten-degrading colony screening approach, a bacterial isolate (2RA3) displaying the highest glutenase activity was selected, characterized and its genome completely sequenced. The identification through 16S rDNA gene sequencing showed a 99,1% similarity to Chryseobacterium taeanense. Hydrolysis of gluten immunogenic peptides (GIP) was further monitored, over a 48-hour period, by colony encapsulation in gliadin-containing microspheres, followed by detection with the G12 anti-GIP monoclonal antibody. Glutenase activity was detected in the extracellular medium of 2RA3 cultures, where gel electrophoresis and gliadin zymography revealed the presence of a ~50 kDa gluten-degrading enzyme. Nano-ESI-Q-TOF of the excised active band identified 7 peptides contained in the protein product predicted for an open reading frame (ORF) in the 2RA3 genome. Based on sequence similarity to the PEP family, the new enzyme was named PEP 2RA3. The PEP 2RA3 coding sequence was PCR-amplified from C. taeanense 2RA3, cloned and expressed in Escherichia coli as a C-terminally His-tagged recombinant protein and purified by Ni-NTA affinity chromatography. The recombinant protein, with predicted molecular mass and isoelectric point of 78.95 kDa and 6.8, respectively, shows PEP activity with standard chromogenic substrates, works optimally at pH 8.0 and 30°C and remains stable at pH 6.0 and 50°C, indicating a potential use in gluten-containing food process applications. The ability of the recombinant enzyme to degrade GIP in beer into smaller peptides was confirmed.

Global Proteome Profiling Reveals Drug-Resistant Traits in Elizabethkingia meningoseptica: An Opportunistic Nosocomial Pathogen.

Elizabethkingia meningoseptica is Gram-negative, rod-shaped opportunistic bacterial pathogen increasingly reported in hospital-acquired outbreaks. This bacterium is well known to thrive in the hospital environment. One of the leading causes of meningitis in pediatric and immune-compromised patients, E. meningoseptica has been noted as a "pathogen of interest" in the context of nosocomial diseases associated with device-related infections in particular. This pathogen's multidrug-resistant phenotype and attendant lack of adequate molecular mechanistic data limit the current approaches for its effective management in hospitals and public health settings. This study provides the global proteome of E. meningoseptica. The reference strain E. meningoseptica ATCC 13253 was used for proteomic analysis using high-resolution Fourier transform mass spectrometry. The study provided translational evidence for 2506 proteins of E. meningoseptica. We identified multiple metallo-ß-lactamases, transcriptional regulators, and efflux transporter proteins associated with multidrug resistance. A protein Car D, which is an enzyme of the carbapenem synthesis pathway, was also discovered in E. meningoseptica. Further, the proteomics data were harnessed for refining the genome annotation. We discovered 39 novel protein-coding genes and corrected four existing translations using proteogenomic workflow. Novel translations reported in this study enhance the molecular data on this organism, thus improving current databases. We believe that the in-depth proteomic data presented in this study offer a platform for accelerated research on this pathogen. The identification of multiple proteins, particularly those involved in drug resistance, offers new future opportunities to design novel and specific antibiotics against infections caused by E. meningoseptica.

Extraction and partial characterisation of antioxidant pigment produced by Chryseobacterium sp. kr6.

Pigments synthesised by Chryseobacterium sp. kr6 growing on feather waste were extracted and characterised. The pigment extract was characterised by KOH test, UV-vis, CIELAB colour system, HPLC-DAD-MS, FTIR and its antioxidant capacity was evaluated. A positive bathochromic shift was observed when kr6 colonies or pigment extracts were subjected to alkaline solution (20% KOH) and a λmax at 450 nm was detected for acetone extracts, although no typical fine structure of carotenoids was detected in the electomagnetic spectra. The HPLC profile of the extracted pigment showed that the compound has three different peaks with λmax near 450 nm. The FTIR analysis shows some principal functional groups from a flexirubin-like molecule. The pigmented compound also presents antioxidant activity evaluated by the scavenging of the ABTS radical.

Effect of hydroxypropyl-ß-cyclodextrin on the cometabolism of phenol and phenanthrene by a novel Chryseobacterium sp.

Cometabolic degradation is an effective method to remove the polycyclic aromatic hydrocarbons (PAHs) with phenol as growth substrate from coal chemical wastewater (CCW). Unfortunately, the toxicity and low solubility of PAHs always restrict their degradation. In this study, Chryseobacterium sp. H202 was firstly isolated from the aerobic segment of CCW. Then, to improve the cometabolic degradation of PAHs, the effects of hydroxypropyl-ß-cyclodextrin (HPCD) were investigated. Phenanthrene removal was accelerated in the presence of phenol; however, the degradation of phenol was inhibited because of the toxicity of phenanthrene. Addition of 50 mg/L HPCD accelerated the degradation of phenol and effectively improved the phenanthrene removal rate by about 55%. Inclusion of HPCD appeared to increase the apparent solubility and reduce the toxicity of phenanthrene, thereby improving the cometabolic degradation of phenol and phenanthrene. Therefore, HPCD can enhance the degradation of phenanthrene with phenol as the growth substrate during CCW treatment.

Diversity and metabolic potential of the microbiota associated with a soil arthropod.

Springtails are important members of the soil fauna and play a key role in plant litter decomposition, for example through stimulation of the microbial activity. However, their interaction with soil microorganisms remains poorly understood and it is unclear which microorganisms are associated to the springtail (endo) microbiota. Therefore, we assessed the structure of the microbiota of the springtail Orchesella cincta (L.) using 16S rRNA gene amplicon sequencing. Individuals were sampled across sites in the field and the microbiota and in particular the endomicrobiota were investigated. The microbiota was dominated by the families of Rickettsiaceae, Enterobacteriaceae and Comamonadaceae and at the genus level the most abundant genera included Rickettsia, Chryseobacterium, Pseudomonas, and Stenotrophomonas. Microbial communities were distinct for the interior of the springtails for measures of community diversity and exhibited structure according to collection sites. Functional analysis of the springtail bacterial community suggests that abundant members of the microbiota may be associated with metabolism including decomposition processes. Together these results add to the understanding of the microbiota of springtails and interaction with soil microorganisms including their putative functional roles.

Chryseobacterium chungangensis sp. nov., a bacterium isolated from soil of sweet gourd garden.

A novel bacterial strain, named MAH-7T, was isolated from a soil sample of a Korean sweet gourd garden and was characterized using a polyphasic approach. Cells were Gram-staining negative, orange colored, non-motile and rod shaped. The strain was aerobic and catalase, oxidase positive, optimum growth temperature and pH were 28-30 °C and 7.0, respectively. On the basis of 16S rRNA gene sequence analysis, strain MAH-7T belongs to the genus Chryseobacterium and is most closely related to Chryseobacterium formosense CC-H3-2T (97.96%) and Chryseobacterium zeae JM-1085T (97.19%). In DNA-DNA hybridization tests, the DNA relatedness between strain MAH-7T and its closest phylogenetic neighbors were below 45.0%. The DNA G+C content was 37.6 mol% and the predominant respiratory quinone was menaquinone-6 (MK-6). Flexirubin-type pigments were found to be present. The major cellular fatty acids were C15:0 iso, C17:0 iso 3OH, C17:1 isoω9c and summed feature 3 (C16:1ω7c and/or C16:1ω6c). The DNA-DNA hybridization results and results of the genotypic analysis in combination with chemotaxonomic and physiological data demonstrated that strain MAH-7T represented a novel species within the genus Chryseobacterium, for which the name Chryseobacterium chungangensis is proposed. The type strain is MAH-7T (= KACC 19293T = CGMCC 1.16232T). The NCBI GenBank accession number for the 16S rRNA gene sequence of strain MAH-7T is KY964274.

Comparing PTR-MS profile of milk inoculated with pure or mixed cultures of spoilage bacteria.

The volatile organic compounds (VOCs) associated with UHT milk (n=8) inoculated with either pure inoculums of Pseudomonas fluorescens (two strains tested) or Chryseobacterium sp., or with mixed cultures of 2 or all 3 of the bacterial strains, and held at 4.5 °C for up to 26 days was measured using proton transfer reaction - mass spectrometry (PTR-MS). The VOCs evolved included a range of carbonyl compounds, alcohols, esters, and acids and had significant qualitative and quantitative differences between the inoculums. Milks inoculated with paired (mixed) bacterial cultures attained patterns similar to the VOC composition of one of the pure inoculums, which could be attributed to the domination of these bacteria within the mixed inoculum. This study will help to characterize the spoilage of milk and provide important insights into understanding the factors that limit the shelf life of milk.