Identification and functional analysis of non-coding regulatory small RNA FenSr3 in Bacillus amyloliquefaciens LPB-18.

Bacillus amyloliquefaciens is an interesting microbe in the food processing and manufacturing industries. Non-coding small RNAs (sRNAs) have been shown to play a crucial role in the physiology and metabolism of bacteria by post-transcriptionally regulating gene expression. This study investigated the function of novel sRNA FenSr3 by constructing fenSr3 deficient strain and complementary strains in B. amyloliquefaciens LPB-18 , which were named LPN-18N and LPB-18P, respectively. The result showed significant differences in fengycin yield between strain LPB -18N and LPB-18P. The production of fengycin was significantly enhanced in B. amyloliquefaciens LPB-18N, compared with that of the strain LPB-18 from 190.908 mg/L to 327.598 mg/L. Moreover, the production of fengycin decreased from 190.464 mg/L to 38.6 mg/L in B . amyloliquefaciens LPB-18P. A comparative transcriptome sequencing was carried out to better understand the complex regulatory mechanism. Transcription analysis revealed that 1037 genes were differentially expressed between B. amyloliquefaciens LPB-18 and B. amyloliquefaciens LPB-18N, including the key regulatory genes in fatty acid, amino acid biosynthesis, and central carbon metabolism, which could provide sufficient quantities of building precursors for fengycin biosynthesis. The biofilm formation and sporulation was also enhanced in the strain LPB-18N, which indicates that FenSr3 could play a vital role in stress resistance and promotes survival in B. amyloliquefaciens. Some sRNAs involved in stress response have been identified in the literature, but their regulatory roles in fengycin production remain unclear. The study will contribute a novel perspective to the regulation mechanism of biosynthesis and the optimization of key metabolites of B. amyloliquefaciens.

Isolation of a potential probiotic strain Bacillus amyloliquefaciensLPB-18 and identification of antimicrobial compounds responsible for inhibition of food-borne pathogens.

This study was carried out to screen a potential probiotic microbe with broad-spectrum antagonistic activity against food-borne pathogens and identify the antimicrobial compounds. Based on morphological and molecular analysis, a new Bacillus strain with the ability to produce effective antimicrobial agents was isolated from the breeding soil of earthworms and identified as having a close evolutionary footprint to Bacillus amyloliquefaciens. The antimicrobial substances produced by B. amyloliquefaciens show effective inhibition of Aspergillus flavus and Fusarium oxysporum in an agar diffusion assay. Antimicrobial agents were identified as a series of fengycin and its isoforms (fengycin A and fengycin B) after being submitted to RT-HPLC and MALDI-TOF MS analyses. To evaluate the probiotic activity of the B. amyloliquefaciens, antibiotic safety and viability of the isolated strain in a simulated gastrointestinal environment were carried out. The safety test result revealed that strain LPB-18 is susceptible to multiple common antibiotics. Moreover, acidic condition and bile salts assay were carried out, and the results revealed that it couble be a potential probiotic microbe B. amyloliquefaciens LPB-18 is good choice for biological strains in agricultural commodities and animal feedstuffs.

Effect of oxidative aging of biochar on relative distribution of competitive adsorption mechanism of Cd2+ and Pb2.

In this study, aged biochar (CCB350 and CCB650) were obtained from pyrolysis of corn stalk biochar (CB350 and CB650) at the degree of 350 °C and 650 °C by artificial oxidation with hydrogen peroxide (H2O2). Also, the mechanism of Pb2+ and Cd2+ on fresh and aged biochars was analyzed qualitatively and quantitatively by batch adsorption experiments combined with characterization. The adsorption isotherm results showed that aging treatment decreased the adsorption capacity of Pb2+ and Cd2+ and inhibited the competitive adsorption behavior of heavy metals. In the single-metal system, precipitation and cation exchange were considered as the main adsorption mechanisms for CB350 and CB650, with a ratio of 40.07-48.23% and 38.04-57.19%, respectively. Competition between Pb2+ and Cd2+ increased the relative contribution of mineral precipitation, but decreased the contribution of cation exchange mechanism. Aging resulted in the rise of the contribution of surface complexation to the adsorption of Pb2+ and Cd2+ on biochars, especially in low-temperature biochars, but weakened the contribution of mineral precipitation to the adsorption. Further, the contribution of other adsorption mechanisms was significantly enhanced for high-temperature aged biochars. These results are important to evaluate its long-term application prospects in the natural environment.

Transcriptome Analysis of Bacillus amyloliquefaciens Reveals Fructose Addition Effects on Fengycin Synthesis.

Fengycin is a lipopeptide produced by Bacillus that has a strong inhibitory effect on filamentous fungi; however, its use is restricted due to poor production and low yield. Previous studies have shown that fengycin biosynthesis in B. amyloliquefaciens was found to be significantly increased after fructose addition. This study investigated the effect of fructose on fengycin production and its regulation mechanism in B. amyloliquefaciens by transcriptome sequencing. According to the RNA sequencing data, 458 genes were upregulated and 879 genes were downregulated. Transcriptome analysis results showed that fructose changed the transcription of amino acid synthesis, fatty acid metabolism, and energy metabolism; alterations in these metabolic pathways contribute to the synthesis of fengycin. In an MLF medium (modified Landy medium with fructose), the expression level of the fengycin operon was two-times higher than in an ML medium (modified Landy medium). After fructose was added to B. amyloliquefaciens, the fengycin-synthesis-associated genes were activated in the process of fengycin synthesis.

Selection of optimal models for predicting growth stress in Artemisia desertorum by comparison of linear regression and multiple neural networks: Take the construction of a green mine in the Bayan Obo mine as an example.

In recent years, more and more countries are focusing on the control of mining sites and the surrounding ecological environment, and the new environmental concept of green mines has been proposed. By investigating the ecological background of a mine site, pollution and ecological imbalances in the mine can be predicted, managed or transformed. This study investigated the effects of rare earth elements on plant growth in the Baotou Bayan Obo Rare Earth Mine and evaluated soil contamination and subsequent remediation through the measured plant height. Using linear regression, BP(Back Propagation) neural networks, GA-BP(Genetic Algorithm- Back Propagation) neural networks, ELM(Extreme Learning Machine) and GA-ELM(Genetic Algorithm- Extreme Learning Machine) model prediction instruments, the different rare earth solution concentrations were set as input values and the heights of Artemisia desertorum, which as the model plant, were set as output values in the prediction. The results showed that the linear regression predicted the standard error of single La(III), Ce(III) solution and compound La(III) + Ce(III) solution for Artemisia desertorum growth stress was on the high side, 7.02%- 8.92%; the efficiency range of each group of models under BP neural network, GA-BP neural network and ELM neural network were 1.15%- 2.53%, 0.85%- 1.28%, 1.76%- 3.53%; while the efficiency range under GA-ELM neural network was 0.59%- 0.68%, with average error values and predicted values close to the true values. Among them, the MAPE of GA-ELM neural network are significantly lower than other models, and the error decreases with increasing concentration of the compound solution. So GA-ELM neural network can be used as an efficient, fast and reasonable optimal model for predicting the growth stress of Artemisia desertorum in Bayan Obo mining area. The experimental results can provide a theoretical basis for assessing the risk of soil rare earth contamination in the area, evaluating the expectation of later remediation, and provide a degree of new ideas for the construction of green mines.

iTRAQ-BASED Proteomic Analysis of the Mechanism of Fructose on Improving Fengycin Biosynthesis in Bacillus Amyloliquefaciens.

Fengycin, as a lipopeptide produced by Bacillus subtilis, displays potent activity against filamentous fungi, including Aspergillus flavus and Soft-rot fungus, which exhibits a wide range of potential applications in food industries, agriculture, and medicine. To better clarify the regulatory mechanism of fructose on fengycin biosynthesis, the iTRAQ-based proteomic analysis was utilized to investigate the differentially expressed proteins of B. amyloliquefaciens fmb-60 cultivated in ML (without fructose) and MLF (with fructose) medium. The results indicated that a total of 811 proteins, including 248 proteins with differential expression levels (162 which were upregulated (fold > 2) and 86, which were downregulated (fold < 0.5) were detected, and most of the proteins are associated with cellular metabolism, biosynthesis, and biological regulation process. Moreover, the target genes' relative expression was conducted using quantitative real-time PCR to validate the proteomic analysis results. Based on the results of proteome analysis, the supposed pathways of fructose enhancing fengycin biosynthesis in B. amyloliquefaciens fmb-60 can be summarized as improvement of the metabolic process, including cellular amino acid and amide, fatty acid biosynthesis, peptide and protein, nucleotide and nucleobase-containing compound, drug/toxin, cofactor, and vitamin; reinforcement of peptide/protein translation, modification, biological process, and response to a stimulus. In conclusion, this study represents a comprehensive and systematic investigation of the fructose mechanism on improving fengycin biosynthesis in B. amyloliquefaciens, which will provide a road map to facilitate the potential application of fengycin or its homolog in defending against filamentous fungi.

Combined effect of cytokine gene polymorphisms on end-stage knee osteoarthritis from Chinese Han population.

The mechanism of osteoarthritis (OA) is not well understood. Cytokines have been implicated in the episode, and there is increasing evidence that the host's cytokine response is genetically determined. We determined the predictive value of IL-634G./C, ICAM-1 469 K/E and IL-10-1082A/G, -819T/C and -592A/C gene polymorphisms on knee OA. The study included 1007 patients with end-stage knee OA and 910 healthy controls. Genomic DNA was prepared from peripheral blood leukocytes. Genotypes and allele frequencies were determined using restriction fragment length polymorphism analysis of polymerase chain reaction products. No significant difference in the IL-10 promoter allele or haplotype frequencies between end-stage knee OA and controls was found. Patients with end-stage knee OA showed a significantly higher prevalence of IL-6-634G/ICAM-1 469E carrier than that in controls (P = 0.017). Results indicate that IL-6-634G/ICAM-1 469E carrier could be associated with increased susceptibility to end-stage knee OA.