Whole genome sequencing and transcriptomics-based characterization of a novel ß-cypermethrin-degrading Gordonia alkanivorans GH-1 isolated from fermented foods.

Beta-cypermethrin (ß-CY) is an organic compound that is widely used as a synthetic pesticide in agriculture and family. Excessive accumulation of ß-CY inevitably causes environmental pollution, which has led to food safety and human health concerns. Identification of microorganisms from food sources that are capable of ß-CY biodegradation may help prevent pollution due to ß-CY accumulation. Here, Gordonia alkanivorans GH-1, which was isolated from the traditional Sichuan fermented food, Pixian Doubanjiang, could not only degrade 82.76% of 50 mg/L ß-CY at 96 h, but also degraded the intermediate degradation products including dibutyl phthalate (DBP), benzoic acid (BA) and phenol (Ph). This bacterial strain, thus, effectively improved the efficiency of removal of ß-CY and its related metabolites, without being limited by toxic intermediates. Whole genome sequencing and transcriptomics analyses have demonstrated that the bacteria affected the transcription of genes related to cell response and material transport under the stress induced by ß-CY, and thereby promoted degradation and transformation of ß-CY. Moreover, a complete pathway of ß-CY degradation is proposed based on the key genes involved in degradation. This study provides important theoretical significance and reference value for eliminating pesticide residues in agricultural products and food to ensure food safety.

Determination of aflatoxin B1 in Pixian Douban based on aptamer magnetic solid-phase extraction.

Aflatoxin B1 (AFB1) is considered as the most prevalent and toxic mycotoxin in food, and is the indispensable index in the monitoring of Pixian Douban, a traditional chinese fermented bean paste from Sichuan. However, the effeciency of AFB1 detection in Pixian Douban is influenced by the traditional extraction, which is usually complex and time consuming. Therefore, an aptamer-based magnetic solid-phase extraction method was designed for the pretreatment of AFB1 in this sample, for which Fe3O4 was synthesized via the solvothermal method and then a Fe3O4@SiO2-NH2 with a core-shell structure was prepared, followed by an AFB1-aptamer attachment. The validation was performed via an enzyme-linked immunosorbent assay and compared with HPLC-MS/MS. The linearity range of this method was 0.5-2.0 ng mL -1 with R 2 of 0.981, and recoveries of AFB1 ranged from 80.19% to 113.92% with RSDs below 7.28% with no significant differences compared to HPLC-MS/MS. The three-time reusability efficiencies of aptamer-MNPs were averaged at 78.24%. The results proved that aptamer-MNPs were high-performance adsorbents for extracting and enriching AFB1, facilitating quick and effective detection of AFB1 in Pixian DouBan samples.

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.

Discrimination of Zanthoxylum bungeanum Maxim through volatile aroma compounds analysis with artificial neural network.

Zanthoxylum bungeanum Maxim (ZBM), a special spice from Chinese different areas, have a widespread variation in quality and price. To avoid the commercial adulteration of ZBM, it is necessary to discriminate them from different areas. As volatile aroma compounds (VAC) have the potential to discriminate ZBM, electronic nose (E-nose) was used to preliminarily discriminate the VAC through sensor response analysis, radar chart analysis, and principal component analysis. Then, Gas chromatography-mass spectrometry (GC-MS) was utilized to identify VAC through hierarchical cluster analysis and quantitative analysis. Finally, artificial neural network (ANN) was employed to assess the accuracy of the discrimination of ZBM. As a result, we found that ZBM could be successfully discriminated between Chinese Sichuan and the other areas. Our findings would provide guidance for evaluating and predicting the variation of VAC of ZBM from different areas in further study. PRACTICAL APPLICATIONS: Zanthoxylum bungeanum Maxim (ZBM) is a traditional and important spice used in Sichuan cuisine especially hotpot, which are famous all over overseas. However, the ZBM from different producing areas bring various flavors, hampering the quality of Sichuan cuisine developing toward to standardization. Therefore, the authors in this work pursuit an effective way to distinguish the ZBM produced in Sichuan rather than in other province. According to the results of the present study, ZBM could be successfully discriminated between Chinese Sichuan and the other producing areas by using E-nose and GC-MS through artificial neural network. These findings would provide the guidance for evaluating the producing areas of ZBM to be whether or not Sichuan, which could offer the practical help in the purchase of the raw material in the supply chain. Besides, these also can be applied to predict the variation of volatile aroma compounds of the ZBM in the further study.

Characterization of deltamethrin degradation and metabolic pathway by co-culture of Acinetobacter junii LH-1-1 and Klebsiella pneumoniae BPBA052.

Deltamethrin and its major metabolite 3-phenoxybenzoic acid (3-PBA) have caused serious threat to the environment as well as human health, yet little is known about their degradation pathways by bacterial co-cultures. In this study, the growth and degradation kinetics of Acinetobacter junii LH-1-1 and Klebsiella pneumoniae BPBA052 during deltamethrin and 3-PBA degradation were established, respectively. When the inoculum proportion of the strains LH-1-1 and BPBA052 was 7.5:2.5, and LH-1-1 was inoculated 24 h before inoculation of strain BPBA052, 94.25% deltamethrin was degraded and 9.16 mg/L of 3-PBA remained within 72 h, which was 20.36% higher and 10.25 mg/L lesser than that in monoculture of LH-1-1, respectively. And the half-life of deltamethrin was shortened from 38.40 h to 24.58 h. Based on gas chromatography-mass spectrometry, 3-phenoxybenzaldehyde, 1,2-benzenedicarboxylic butyl dacyl ester, and phenol were identified as metabolites during deltamethrin degradation in co-culture. This is the first time that a co-culture degradation pathway of deltamethrin has been proposed based on these identified metabolites. Bioremediation of deltamethrin-contaminated soils with co-culture of strains LH-1-1 and BPBA052 significantly enhanced deltamethrin degradation and 3-PBA removal. This study provides a platform for further studies on deltamethrin and 3-PBA biodegradation mechanism in co-culture, and it also proposes a promising approach for efficient bioremediation of environment contaminated by pyrethroid pesticides and their associated metabolites.

Improved protease activity of Pixian broad bean paste with cocultivation of Aspergillus oryzae QM-6 and Aspergillus niger QH-3

BACKGROUND: The preparation of broad bean koji is a key process in the production of Pixian broad bean paste (PBP). Protease is essential for the degradation of proteins during PBP fermentation. To obtain broad bean koji with high protease activity using the cocultivated strains of Aspergillus oryzae QM-6 (A. oryzae QM-6) and Aspergillus niger QH-3 (A. niger QH-3), the optimization of acid and neutral protease activities was carried out using Box­Behnken design with response surface methodology (RSM). RESULTS: The optimum conditions were found to be as follows: inoculation proportion (X1), 3:1 (A. oryzae QM-6: A. niger QH-3, w/w); culture temperature (X2), 33°C; inoculum size (X3), 0.5% (w/w); incubation time (X4), 5 d. The acid and neutral protease activities were 605.2 ± 12.4 U/g and 1582.9 ± 23.7 U/g, respectively, which were in good agreement with the predicted values. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis profiles revealed that the broad bean koji extracellular proteins in the case of cocultivation were richer compared to those in the case of A. oryzae QM-6 or A. niger QH-3 strain only. In addition, the free amino acids (FAAs) in the fermentation product were 55% higher in the cocultivation process than in that involving only A. oryzae QM-6, further confirming the diversity of proteases in the fermentation products. CONCLUSIONS: The optimal conditions of koji-making in PBP were obtained using RSM. The cocultivation of A. oryzae and A. niger increases the overall enzyme activities in the culture medium and the FAAs content, which would thus have potential application in the PBP industry.

Efficient biodegradation of 3-phenoxybenzoic acid and pyrethroid pesticides by the novel strain Klebsiella pneumoniae BPBA052.

A novel Klebsiella pneumoniae strain (BPBA052) capable of degrading 3-phenoxybenzoic acid (3-PBA) was isolated from soybean rhizosphere soil. The strain was obtained by screening after enrichment, isolation, and purification using 3-PBA as the sole carbon and energy source. It could degrade 96.37% of 3-PBA (100 mg/L) within 72 h, and its growth and 3-PBA degradation followed kinetics models of logistic growth (XBPBA052 = 0.0883 × e0.0947t / [1 - 0.0792 × (1 - 0.0883 × e0.0947t)]; µm = 0.0947 h-1, X0 = 0.0883, and Xm = 1.1145) and first-order degradation (CBPBA052 = 101.8194 × e-0.0403t, k = 0.0403, t1/2 = 17.22 h), respectively. Based on Box-Behnken response surface analysis, the optimal temperature, pH, and 3-PBA concentration for K. pneumoniae BPBA052 were 35.01 °C, 7.77, and 150 mg/L, respectively. Moreover, pyrethroid pesticides (PPs) (such as ß-cypermethrin, permethrin, bifenthrin, deltamethrin, and fenvalerate) and 3-PBA metabolites (including phenol, catechol, and protocatechuate) were efficiently utilized by BPBA052. We propose a novel microbial metabolic pathway for 3-PBA, based on metabolite identification; enzyme-degrading activity; and cloning of the phenol hydroxylase, catechol 1,2-dioxygenase, and protocatechuate 3,4-dioxygenase genes. This study provides a fundamental platform for further studies to reveal the mechanism of biodegradation of 3-BPA and show K. pneumoniae BPBA052 as a potential microbial resource for bioremediation of environments polluted with 3-PBA or PPs.

Screening of a beta-cypermethrin-degrading bacterial strain Brevibacillus parabrevis BCP-09 and its biochemical degradation pathway.

A novel beta-cypermethrin (Beta-CP)-degrading strain isolated from activated sludge was identified as Brevibacillus parabrevis BCP-09 based on its morphological and physio-biochemical characteristics, and 16S rRNA gene analysis. Strain BCP-09 could effectively degrade Beta-CP at pH 5.0-9.0, 20-40 °C, and 10-500 mg L-1 Beta-CP. Under optimal conditions (pH 7.41, 38.9 °C, 30.9 mg L-1 Beta-CP), 75.87% Beta-CP was degraded within 3 days. Beta-CP degradation (half-life, 33.45 h) and strain BCP-09 growth were respectively described using first-order-kinetic and logistic-kinetic models. Seven metabolites were detected by high-performance liquid chromatography and gas chromatography-mass spectrometry- methyl salicylate, catechol, phthalic acid, salicylic acid, 3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropanecarboxylic acid, 3-phenoxybenzaldehyde, and 3-phenoxybenzoic acid (3-PBA). The major Beta-CP metabolite, 3-PBA was further degraded into phenol, benzoic acid, and 4-methylhexanoic acid. BCP-09 also degraded aromatic compounds such as phenol, catechol, and protocatechuic acid. Beta-CP appears to be mainly degraded into 3-PBA, which is continuously degraded into smaller benzene or chain compounds. Thus, strain BCP-09 could form a complete degradation system for Beta-CP and might be considered a promising strain for application in the bioremediation of environments and agricultural products polluted by Beta-CP.