A novel strain of Stenotrophomonas acidaminiphila produces thermostable alkaline peptidase on agro-industrial wastes: process optimization, kinetic modeling and scale-up.

Bacterial alkaline peptidases, especially from Bacillus species, occupy the frontline in global enzyme market, albeit with poor production economics. Here, we report the deployment of response surface methodology approximations to optimize fermentation parameters for enhanced yield of alkaline peptidase by the non-Bacillus bacterium; Stenotrophomonas acidaminiphila. Shake flask production under optimized conditions was scaled up in a 5-L bench-scale bioreactor. Logistic and modified Gompertz models revealed significant fits for biomass formation, total protein, and substrate consumption models. Maximum specific growth rate (µmax = 0.362 h-1) of the bacterium in the optimized medium did not differ significantly from those in Luria-Bertani and trypticase soy broths. The aqueous two-phase system-purified 45.7 kDa alkaline protease retained 83% activity which improved with increasing sodium dodecyl sulfate concentration thus highlighting potential laundry application. Maximum enzyme activity occurred at 75ºC and pH 10.5 but was inhibited by 5 mM phenyl-methyl-sulfonyl fluoride suggesting a serine-protease nature.

High-performance bacterium-enhanced dual-compartment microbial fuel cells for simultaneous treatment of food waste oil and copper-containing wastewater.

Microbial fuel cells (MFCs) use the metabolic actions of microorganisms in an anode chamber to convert the chemical energy from wastewater into electrical energy. To improve the MFC power generation performance and chemical oxygen demand (COD) removal efficiency, Stenotrophomonas acidaminiphila was added to the anode chamber of a dual-compartment MFC. In this process, Stenotrophomonas acidaminiphila promotes the degradation of macromolecules such as bis(2-ethylhexyl) phthalate in food waste oil. Additionally, the generated electrical energy reduced Cu2+ in the copper-containing wastewater in the cathode chamber to Cu monomers. The maximum power density of the MFC was 49.5 ± 3.5 mW/m2, the maximum removal efficiencies of COD and Cu2+ were 63.5 ± 5.8% and 96.5 ± 1.0%, respectively, and Cu2+ was reduced to brick-red Cu monomers. This study provides insights into the simultaneous implementation of food waste oil treatment and metal resource recovery.

Characteristics of Aflatoxin B1 Degradation by Stenotrophomonas acidaminiphila and It's Combination with Black Soldier Fly Larvae.

Aflatoxin B1 (AFB1) is a common mycotoxin contaminant in cereals that causes severe economic losses and serious risks to the health of humans and animals. In this paper, we investigated the characteristics of AFB1 degradation by black soldier fly larvae (BSFL) combined with commensal intestinal microorganisms. Germ-free BSFL and non-sterile BSFL were reared on peanut meal spiked with AFB1 for 10 days. The result showed that germ-free BSFL and non-sterile BSFL could achieve 31.71% and 88.72% AFB1 degradation, respectively, which indicated the important role of larvae gut microbiota in AFB1 degradation. Furthermore, twenty-five AFB1-degrading bacteria were isolated from BSFL gut, and S. acidaminiphila A2 achieved the highest AFB1 degradation, by 94%. When S. acidaminiphila A2 was re-inoculated to BSFL, the detrimental effect of AFB1 on the growth performance of BSFL was alleviated, and complete AFB1 degradation in peanut meal was obtained. In conclusion, the present study may provide a strategy to degrade AFB1 in feedstuff through bioconversion with BSFL in combination with gut-originated AFB1-degrading bacteria, while providing a sustainable insect protein and fat source to animals.

Effects of Free or Immobilized Bacterium Stenotrophomonas acidaminiphila Y4B on Glyphosate Degradation Performance and Indigenous Microbial Community Structure.

The overuse of glyphosate has resulted in serious environmental contamination. Thus, effective techniques to remove glyphosate from the environment are required. Herein, we isolated a novel strain Stenotrophomonas acidaminiphila Y4B, which completely degraded glyphosate and its major metabolite aminomethylphosphonic acid (AMPA). Y4B degraded glyphosate over a broad concentration range (50-800 mg L-1), with a degradation efficiency of over 98% within 72 h (50 mg L-1). Y4B degraded glyphosate via the AMPA pathway by cleaving the C-N bond, followed by degradation of AMPA and subsequent metabolism. Y4B demonstrated strong competitiveness and substantially accelerated the degradation of glyphosate in different soils, degrading 71.93 and 89.81% of glyphosate (400 mg kg-1) within 5 days in sterile and nonsterile soils, respectively. The immobilized cells of Y4B were more efficient than their free cells and they displayed excellent biodegradation efficiency in a sediment-water system. Taken together, Y4B is an ideal degrader for the bioremediation of glyphosate-contaminated sites.

Isolation of Dibutyl Phthalate-Degrading Bacteria and Its Coculture with Citrobacter freundii CD-9 to Degrade Fenvalerate.

Continued fenvalerate use has caused serious environmental pollution and requires large-scale remediation. Dibutyl phthalate (DBP) was discovered in fenvalerate metabolites degraded by Citrobacter freundii CD-9. Coculturing is an effective method for bioremediation, but few studies have analyzed the degradation pathways and potential mechanisms of cocultures. Here, a DBP-degrading strain (BDBP 071) was isolated from soil contaminated with pyrethroid pesticides (PPs) and identified as Stenotrophomonas acidaminiphila. The optimum conditions for DBP degradation were determined by response surface methodology (RSM) analysis to be 30.9 mg/l DBP concentration, pH 7.5, at a culture temperature of 37.2°C. Under the optimized conditions, approximately 88% of DBP was degraded within 48 h and five metabolites were detected. Coculturing C. freundii CD-9 and S. acidaminiphila BDBP 071 promoted fenvalerate degradation. When CD-9 was cultured for 16 h before adding BDBP 071, the strain inoculation ratio was 5:5 (v/v), fenvalerate concentration was 75.0 mg/l, fenvalerate was degraded to 84.37 ± 1.25%, and DBP level was reduced by 5.21 mg/l. In addition, 12 fenvalerate metabolites were identified and a pathway for fenvalerate degradation by the cocultured strains was proposed. These results provide theoretical data for further exploration of the mechanisms used by this coculture system to degrade fenvalerate and DBP, and also offer a promising method for effective bioremediation of PPs and their related metabolites in polluted environments.

Genome-wide analysis reveals the emergence of multidrug resistant Stenotrophomonas acidaminiphila strain SINDOREI isolated from a patient with sepsis.

Stenotrophomonas acidaminiphila, the most recent reported species in genus Stenotrophomonas, is a relatively rare bacteria and is an aerobic, glucose non-fermentative, Gram-negative bacterium. However, little information of S. acidaminiphila is known to cause human infections. In this research, we firstly reported a multidrug-resistant strain S. acidaminiphila SINDOREI isolated from the blood of a patient with sepsis, who was dead of infection eventually. The whole genome of strain SINDOREI was sequenced, and genome comparisons were performed among six closely related S. acidaminiphila strains. The core genes (2,506 genes) and strain-specific genes were identified, respectively, to know about the strain-level diversity in six S. acidaminiphila stains. The presence of a unique gene (narG) and essential genes involved in biofilm formation in strain SINDOREI are important for the pathogenesis of infections. Strain SINDOREI was resistant to trimethoprim/sulfamethoxazole, ciprofloxacin, ofloxacin, cefepime, ceftazidime, and aztreonam. Several common and specific antibiotic resistance genes were identified in strain SINDOREI. The presence of two sul genes and exclusive determinants GES-1, aadA3, qacL, and cmlA5 is responsible for the resistance to multidrug. The virulence factors and resistance determinants can show the relationship between the phenotype and genotype and afford potential therapeutic strategies for infections.

Biotransformation of chlorothalonil by strain Stenotrophomonas acidaminiphila BJ1 isolated from farmland soil.

Chlorothalonil is a widely used fungicide, but the contamination of soil and water environments by this chemical causes potential threats to biodiversity. Given the metabolic potential of soil microorganisms, there is a need for developing microbiological approaches to degrade persistent compounds, such as chlorothalonil, in contaminated sites. Here in this study, we isolated a bacterial strain (namely, BJ1) capable of degrading chlorothalonil from a chlorothalonil-contaminated farmland soil in the Shandong Province, China. Using 16S rDNA gene sequencing, morphological and biological characteristics, we identified the strain BJ1 as Stenotrophomonas acidaminiphila. The strain BJ1 uses chlorothalonil as a sole carbon source. At initial concentrations of 50, 100, 200 and 300 mg l-1, it degraded 91.5%, 89.4%, 86.5% and 83.5% of chlorothalonil after 96 h of inoculation under optimum conditions (30°C and pH 7.0). Two metabolites, methyl-2,5,6-trichloro-3-cyano-4-methoxy-benzoate and methyl-3-cyano-2,4,5,6-tetrachlorobenzoate, were detected and identified based on HPLC-MS analysis, which suggests that the strain BJ1 metabolized chlorothalonil through the hydroxylation of chloro-group and hydration of cyano-group. The results of this study highlight the great potential for this bacterium to be used in chlorothalonil pollution remediation.

Genome Sequencing and Comparative Analysis of Stenotrophomonas acidaminiphila Reveal Evolutionary Insights Into Sulfamethoxazole Resistance.

Stenotrophomonas acidaminiphila is an aerobic, glucose non-fermentative, Gram-negative bacterium that been isolated from various environmental sources, particularly aquatic ecosystems. Although resistance to multiple antimicrobial agents has been reported in S. acidaminiphila, the mechanisms are largely unknown. Here, for the first time, we report the complete genome and antimicrobial resistome analysis of a clinical isolate S. acidaminiphila SUNEO which is resistant to sulfamethoxazole. Comparative analysis among closely related strains identified common and strain-specific genes. In particular, comparison with a sulfamethoxazole-sensitive strain identified a mutation within the sulfonamide-binding site of folP in SUNEO, which may reduce the binding affinity of sulfamethoxazole. Selection pressure analysis indicated folP in SUNEO is under purifying selection, which may be owing to long-term administration of sulfonamide against Stenotrophomonas.

Degradation of fipronil by Stenotrophomonas acidaminiphila isolated from rhizospheric soil of Zea mays.

Fipronil is a widely used insecticide in agriculture and can cause potential health hazards to non-target soil invertebrates and nearby aquatic systems. In the present study, a fipronil degrading bacterium was isolated from fipronil contaminated soil, i.e. rhizospheric zone of Zea mays. Morphological, biochemical and molecular characterization of strain indicated that it clearly belongs to Stenotrophomonas acidaminiphila (accession no. KJ396942). A three-factor Box-Behnken experimental design combined with response surface modeling was employed to predict the optimum conditions for fipronil degradation. The optimum pH, temperature and total inocula biomass for the degradation of fipronil were 7.5, 35 °C and 0.175 g L-1, respectively. The bacterial strain was able to metabolize 25 mg L-1 fipronil with 86.14 % degradation in Dorn's broth medium under optimum conditions. Metabolites formed as a result of fipronil degradation were characterized with gas liquid chromatograph. A novel fipronil degradation pathway was proposed for S. acidaminiphila on the basis of metabolites formed. Non-sterilized soil inoculated with S. acidaminiphila was found to follow first order kinetics with a rate constant of 0.046 d-1. Fipronil sulfone, sulfide and amide were formed as the metabolites and were degraded below the quantifiable limit after 90 days of time period. Given the high fipronil degradation observed in the present study, S. acidaminiphila may have potential for use in bioremediation of fipronil contaminated soils.