Bioinformatics analysis of gene bhsA and its role in Ca2+ -treated Escherichia coli.

One of the commonly employed methods in molecular biology is to utilize calcium chloride to treat Escherichia coli for the preparation of competent cells to facilitate foreign gene expression. However, the molecular mechanisms underlying Ca2+ mediation of competent cell formation and identification of the key genes involved in the process remain unclear. In previous studies, the combined analysis of transcriptomics and proteomics revealed bhsA as one of the crucial genes. The gene ontology functional annotation of bhsA identified it as a member of the YhcN family encoding an outer membrane protein that confers resistance to various stresses. The IPR0108542 domain found within the protein plays a significant role in stress response and biofilm formation in E. coli. Analysis of the STRING database showed that the proteins interacting with bhsA are primarily involved in biofilm formation and stress resistance. Using the RED homologous recombination method, a bhsA deletion strain was constructed to verify its role in E. coli genetic transformation. Although the mutant strain showed no significant differences in morphology or growth trend when compared to the wild-type strain, its transformation efficiency decreased by 1.14- and 1.64-fold with plasmids pUC19 and pET-32a. Furthermore, the 1-N-phenylnaphthylamine assay indicated a 1.71-fold reduction in cell membrane permeability in the mutant strain.

Role of cspA on the Preparation of Escherichia coli Competent Cells by Calcium Chloride Method.

One of the fundamental techniques in genetic engineering is the creation of Escherichia coli competent cells using the CaCl2 method. However, little is known about the mechanism of E. coli competence formation. We have previously found that the cspA gene may play an indispensable role in the preparation of E. coli DH5α competent cells through multiomics analysis. In the present study, the cellular localization, physicochemical properties, and function of the protein expressed by the cspA gene were analyzed. To investigate the role of the cspA gene in E. coli transformation, cspA-deficient mutant was constructed by red homologous recombination. The growth, transformation efficiency, and cell morphology of the cspA-deficient strain and E. coli were compared. It was found that there were no noticeable differences in growth and morphology between E. coli and the cspA-deficient strain cultured at 37°C, but the mutant exhibited increased transformation efficiencies compared to E. coli DH5α for plasmids pUC19, pET-32a, and p1304, with enhancements of 2.23, 2.24, and 3.46 times, respectively. It was proved that cspA gene is an important negative regulatory gene in the CaCl2 preparation of competent cells.

Comparative transcriptomics revealed the mechanism of Stenotrophomonas rhizophila JC1 response and biosorption to Pb2.

Nowadays, there is limited research focusing on the biosorption of Pb2+ through microbial process, particularly at the level of gene expression. To overcome this knowledge gap, we studied the adsorption capacity of Stenotrophomonas rhizophila JC1 to Pb2+, and investigated the physiological mechanism by means of SEM, EDS, FTIR, membrane permeability detection, and investigated the molecular mechanism through comparative transcriptomics. The results showed that after 16 h of cultivation, the biosorption capacity of JC1 for 100 mg/L of Pb2+ reached at 79.8%. The main mechanism of JC1 adsorb Pb2+ is via intracellular accumulation, accounting for more than 90% of the total adsorption. At the physiological level, Pb2+ can precipitate with anion functional groups (e.g., -OH, -NH) on the bacterial cell wall or undergo replacement reaction with cell component elements (e.g., Si, Ca) to adsorb Pb2+ outside of the cell wall, thus accomplishing extracellular adsorption of Pb2+ by strains. Furthermore, the cell membrane acts as a "switch" that inhibits the entry of metal ions into the cell from the plasma membrane. At the molecular level, the gene pbt specificity is responsible for the adsorption of Pb2+ by JC1. In addition, phosphate permease is a major member of the ABC transporter family involved in Pb2+, and czcA/cusA or Co2+/Mg2+ efflux protein plays an important role in the efflux of Pb2+ in JC1. Further, cellular macromolecule biosynthesis, inorganic cation transmembrane transport, citrate cycle (TCA) and carbon metabolism pathways all play crucial roles in the response of strain JC1 to Pb2+ stress.

Protection of primary cilia is an effective countermeasure against the impairment of osteoblast function induced by simulated microgravity.

The molecular mechanism for the microgravity-induced decrease in bone formation remains unclear and there is a lack of effective specific preventative therapies. We recently reported that primary cilia of osteoblasts became shorter and even disappeared when the cells were exposed to random positioning machine (RPM)-simulated microgravity and that the microgravity-induced loss of osteogenic potential of osteoblasts could be attenuated when the resorption of primary cilia was prevented by treatment with 0.1 µM cytochalasin D. In the current study, it was further found that the loss of the osteogenic capacity of rat calvarial osteoblasts (ROBs) was associated with the inhibition of the BMP-2/Smad1/5/8 signalling pathway, of which most of the signalling proteins including BMP-2, BMPRII, Smad1/5/8 and p-Smad1/5/8 were found localized to primary cilia. Accompanying the resorption of primary cilia following the cells being exposed to simulated microgravity, the expression levels of these signalling proteins were reduced significantly. Furthermore, the expression of miRNA-129-3p, a microRNA previously reported to control cilium biogenesis, was found to be reduced quickly and changed in a similar tendency with the length of primary cilia. Moreover, overexpression of miRNA-129-3p in ROBs significantly attenuated microgravity-induced inhibition of BMP-2 signalling and loss of osteogenic differentiation and mineralization. These results indicated the important role of miRNA-129-3p in microgravity-induced resorption of primary cilia of osteoblasts and the potential of replenishing the miRNA-129-3p as an effective countermeasure against microgravity-induced loss of primary cilia and impairment of osteoblast function.

Culturable endophytic fungi community structure isolated from Codonopsis pilosula roots and effect of season and geographic location on their structures.

BACKGROUND: Rhizosphere soil physicochemical, endophytic fungi have an important role in plant growth. A large number of endophytic fungi play an indispensable role in promoting plant growth and development, and they can provide protection for host plants by producing a variety of secondary metabolites to resist and inhibit plant pathogens. Due to the terrain of Gansu province is north-south and longitudinal, different climatic conditions, altitude, terrain and growth environment will affect the growth of Codonopsis pilosula, and the changes in these environmental factors directly affect the quality and yield of C. pilosula in different production areas. However, In C. pilosula, the connection between soil nutrients, spatiotemporal variation and the community structure of endophytic fungi isolated from C. pilosula roots has not been well studied. RESULTS: Seven hundred six strains of endophytic fungi were obtained using tissue isolation and the hyphaend-purification method from C. pilosula roots that picked at all seasons and six districts (Huichuan, HC; Longxi, LX; Zhangxian, ZX; Minxian, MX; Weiyuan, WY; and Lintao, LT) in Gansu Province, China. Fusarium sp. (205 strains, 29.04%), Aspergillus sp. (196 strains, 27.76%), Alternaria sp. (73 strains, 10.34%), Penicillium sp. (58 strains, 8.22%) and Plectosphaerella sp. (56 strains, 7.93%) were the dominant genus. The species composition differed from temporal and spatial distribution (Autumn and Winter were higher than Spring and Summer, MX and LT had the highest similarity, HC and LT had the lowest). physical and chemical of soil like Electroconductibility (EC), Total nitrogen (TN), Catalase (CAT), Urease (URE) and Sucrase (SUC) had significant effects on agronomic traits of C. pilosula (P < 0.05). AK (Spring and Summer), TN (Autumn) and altitude (Winter) are the main driving factors for the change of endophytic fungal community. Moreover, geographic location (such as altitude, latitude and longitude) also has effects on the diversity of endophytic fungi. CONCLUSIONS: These results suggested that soil nutrients and enzyme, seasonal variation and geographical locations have an impact on shaping the community structure of culturable endophytic fungi in the roots of C. pilosula and its root traits. This suggests that climatic conditions may play a driving role in the growth and development of C. pilosula.

Cloning, expression, and bioinformatics analysis of heavy metal resistance-related genes fd-I and fd-II from Acidithiobacillus ferrooxidans.

Five heavy metals were introduced into the bacterial heavy metal resistance tests. The results showed that apparent inhibition effects of Cd2+ and Cu2+ on the growth of Acidithiobacillus ferrooxidans BYSW1 occurred at high concentrations (>0.04 mol l-1). Significant differences (P < 0.001) were both noticed in the expression of two ferredoxin-encoding genes (fd-I and fd-II) related to heavy metal resistance in the presence of Cd2+ and Cu2+ . When exposed to 0.06 mol l-1 Cd2+, the relative expression levels of fd-I and fd-II were about 11 and 13 times as much as those of the control, respectively. Similarly, exposure to 0.04 mol l-1 Cu2+ caused approximate 8 and 4 times higher than those of the control, respectively. These two genes were cloned and expressed in Escherichia coli, and the structures, functions of two corresponding target proteins, i.e. Ferredoxin-I (Fd-I) and Ferredoxin-II (Fd-II), were predicted. The recombinant cells inserted by fd-I or fd-II were more resistant to Cd2+ and Cu2+ compared with wild-type cells. This study was the first investigation regarding the contribution of fd-I and fd-II to enhancing heavy metal resistance of this bioleaching bacterium, and laid a foundation for further elucidation of heavy metal resistance mechanisms caused by Fd.

The function of the gene loiP in transformation efficiency and outer membrane permeability change of Escherichia coli treated by Ca2+ ions.

The preparation of Escherichia coli competent cells by calcium chloride is a common method in molecular biology, but the mechanism is poorly understood. In a previous study, using transcriptomics and proteomics approaches, we found that the expression pattern of the gene loiP was upregulated by CaCl2. In order to investigate the function of the loiP gene in Ca2+- mediated formation of competent cells of E. coli DH5α, the loiP gene deletion strains were constructed by the lambda-derived Red homologous recombination technique. Then, the cell morphology, transformation efficiency, and cell membrane changes of the competent cells of the mutant strain were further explored. Compared with the wild-type E. coli DH5α, the mutant strains have no significant differences in the morphology, growth characteristics, and the permeability of the intracellular membrane. However, the transformation efficiencies of the mutant strains to plasmids of different sizes were significantly reduced, and the permeability of the outer membrane decreased by 2.94 times. These results indicate that the deletion of gene loiP may directly affect the transformation efficiency and outer membrane permeability of E. coli competent cells.

Identification of Key Genes during Ca2+-Induced Genetic Transformation in Escherichia coli by Combining Multi-Omics and Gene Knockout Techniques.

The molecular mechanism of the Ca2+-mediated formation of competent cells in Escherichia coli remains unclear. In this study, transcriptome and proteomics techniques were used to screen genes in response to Ca2+ treatment. A total of 333 differentially expressed genes (317 upregulated and 16 downregulated) and 145 differentially expressed proteins (54 upregulated and 91 downregulated) were obtained. These genes and proteins are mainly enriched in cell membrane components, transmembrane transport, and stress response-related functional terms. Fifteen genes with these functions, including yiaW, ygiZ, and osmB, are speculated to play a key role in the cellular response to Ca2+. Three single-gene deletion strains were constructed with the Red homologous recombination method to verify its function in genetic transformation. The transformation efficiencies of yiaW, ygiZ, and osmB deletion strains for different-size plasmids were significantly increased. None of the three gene deletion strains changed in size, which is one of the main elements of microscopic morphology, but they exhibited different membrane permeabilities and transformation efficiencies. This study demonstrates that Ca2+-mediated competence formation in E. coli is not a simple physicochemical process and may involve the regulation of genes in response to Ca2+. This study lays the foundation for further in-depth analyses of the molecular mechanism of Ca2+-mediated transformation. IMPORTANCE Using transcriptome and proteome techniques and association analysis, we identified several key genes involved in the formation of Ca2+-mediated E. coli DH5α competent cells. We used Red homologous recombination technology to construct three single-gene deletion strains and found that the transformation efficiencies of yiaW, ygiZ, and osmB deletion strains for different-size plasmids were significantly increased. These results proved that the genetic transformation process is not only a physicochemical process but also a reaction process involving multiple genes. These results suggest ways to improve the horizontal gene transfer mechanism of foodborne microorganisms and provide new ideas for ensuring the safety of food preservation and processing.

Insights into the Structures, Inhibitors, and Improvement Strategies of Glucose Oxidase.

Glucose oxidase, which uses molecular oxygen as an electron acceptor to specifically catalyze the conversion of ß-d-glucose to gluconic acid and hydrogen peroxide (H2O2), has been considered an important enzyme in increasing environmental sustainability and food security. However, achieving the high yield, low price and high activity required for commercial viability remains challenging. In this review, we first present a brief introduction, looking at the sources, characteristics, catalytic process, and applications of glucose oxidase. Then, the predictive structures of glucose oxidase from two different sources are comparatively discussed. We summarize the inhibitors of glucose oxidase. Finally, we highlight how the production of glucose oxidase can be improved by optimizing the culture conditions and microbial metabolic engineering.

Molecular mechanisms of heavy metals resistance of Stenotrophomonas rhizophila JC1 by whole genome sequencing.

In this study, a higher metal ions-resistant bacterium, Stenotrophomonas rhizophila JC1 was isolated from contaminated soil in Jinchang city, Gansu Province, China. The Pb2+ (120 mg/L) and Cu2+ (80 mg/L) removal rate of the strain reached at 76.9% and 83.4%, respectively. The genome comprises 4268161 bp in a circular chromosome with 67.52% G + C content and encodes 3719 proteins. The genome function analysis showed czc operon, mer operon, cop operon, arsenic detoxification system in strain JC1 were contributed to the removal of heavy metals. Three efflux systems (i.e., RND, CDF, and P-ATPase) on strain JC1 genome could trigger the removal of divalent cations from cells. cAMP pathway and ABC transporter pathway might be involved in the transport and metabolism of heavy metals. The homology analysis exhibited multi-gene families such as ABC transporters, heavy metal-associated domain, copper resistance protein, carbohydrate-binding domain were distributed across 410 orthologous groups. In addition, heavy metal-responsive transcription regulator, thioredoxin, heavy metal transport/detoxification protein, divalent-cation resistance protein CutA, arsenate reductase also played important roles in the heavy metals adsorption and detoxification process. The complete genome data provides insight into the exploration of the interaction mechanism between microorganisms and heavy metals.

Soil Organic Matter and Total Nitrogen Reshaped Root-Associated Bacteria Community and Synergistic Change the Stress Resistance of Codonopsis pilosula.

The stress resistance of medicinal plants is essential to the accumulation of pharmacological active ingredients, but the regulation mechanism of biological factors and abiotic factors on medicinal plants is still unclear. To investigate the mechanism of soil nutrient and microecology on the stress resistance of C. pilosula, rhizosphere soil and roots were collected across the four seasons in Minxian, Gansu, and their physicochemical properties, as well as root-associated microorganisms, were examined. The results showed that the bacterial α-diversity indexes increased in the endosphere and rhizosphere from summer to autumn. At the same time, the community composition and function changed considerably. The stability of the endophytic bacterial community was higher than that rhizospheric bacteria, and the complexity of the endophytic bacterial community was lower than rhizospheric bacteria. Soil organic matter (OM), water content (WC), total potassium (TK), and total nitrogen (TN) have been identified as the key factors affecting bacterial community diversity and stress resistance of C. pilosula. WC, TN, and OM showed significant differences from summer to autumn (P < 0.5). Four key soil physiochemical factors changed significantly between seasons (P < 0.01). TN and OM change the stress resistance of C. pilosula mainly by changing the activity of antioxidant enzymes. Changes of OM and endophytic bacterial diversity affect the accumulation of soluble sugars to alter stress resistance. These four key soil physicochemical factors significantly influenced the diversity of endophytic bacteria. WC and OM were identified as the most important factors for endophytic and rhizospheric bacteria, respectively. This study provided the research basis for the scientific planting of C. pilosula.

Paraphoma chrysanthemicola Affects the Carbohydrate and Lobetyolin Metabolism Regulated by Salicylic Acid in the Soilless Cultivation of Codonopsis pilosula.

Paraphoma chrysanthemicola, an endophytic fungus isolated from the roots of Codonopsis pilosula, influences salicylic acid (SA) levels. The interaction mechanism between SA and P. chrysanthemicola within C. pilosula remains elusive. To elucidate this, an experiment was conducted with four treatments: sterile water (CK), P. chrysanthemicola (FG), SA, and a combination of P. chrysanthemicola with salicylic acid (FG+SA). Results indicated that P. chrysanthemicola enhanced plant growth and counteracted the growth inhibition caused by exogenous SA. Physiological analysis showed that P. chrysanthemicola reduced carbohydrate content and enzymatic activity in C. pilosula without affecting total chlorophyll concentration and attenuated the increase in these parameters induced by exogenous SA. Secondary metabolite profiling showed a decrease in soluble proteins and lobetyolin levels in the FG group, whereas SA treatment led to an increase. Both P. chrysanthemicola and SA treatments decreased antioxidase-like activity. Notably, the FG group exhibited higher nitric oxide (NO) levels, and the SA group exhibited higher hydrogen peroxide (H2O2) levels in the stems. This study elucidated the intricate context of the symbiotic dynamics between the plant species P. chrysanthemicola and C. pilosula, where an antagonistic interaction involving salicylic acid was prominently observed. This antagonism was observed in the equilibrium between carbohydrate metabolism and secondary metabolism. This equilibrium had the potential to engage reactive oxygen species (ROS) and nitric oxide (NO).

Degradation of petroleum hydrocarbon contaminants by Rhodococcus erythropolis KB1 synergistic with alfalfa (Medicago sativa L.).

Petroleum hydrocarbons are a stubborn pollutant that is difficult to degrade globally, and plant-microbial degradation is the main way to solve this type of pollutant. In this study, the physiological and ecological responses of alfalfa to petroleum hydrocarbons in different concentrations of petroleum hydrocarbon-contaminated soil with KB1 (Rhodococcus erythropolis) were analyzed and determined by laboratory potting techniques. The growth of alfalfa (CK) and alfalfa with KB1 (JZ) in different concentrations of petroleum hydrocarbons contaminated soil was compared and analyzed. The results of the CK group showed that petroleum hydrocarbons could significantly affect the activity of alfalfa antioxidant enzyme system, inhibit the development of alfalfa roots and the normal growth of plants, especially in the high-concentration group. KB1 strain had the ability to produce IAA, form biofilm, fix nitrogen, produce betaine and ACC deaminase, and the addition of KB1 could improve the growth traits of alfalfa in the soil contaminated with different concentrations of petroleum hydrocarbons, the content of soluble sugars in roots, and the stress resistance and antioxidant enzyme activities of alfalfa. In addition, the degradation kinetics of the strain showed that the degradation rate of petroleum could reach 75.2% after soaking with KB1. Furthermore, KB1 can efficiently degrade petroleum hydrocarbons in advance and significantly alleviate the damage of high concentration of petroleum hydrocarbons to plant roots. The results showed that KB1 strains and alfalfa plants could effectively enhance the degradation of petroleum hydrocarbons, which provided new ideas for improving bioremediation strategies.

Exopolysaccharides of Serratia fonticola CPSE11 can alleviate the toxic effect of Cd2+ on Codonopsis pilosula.

In order to study the detoxification effect of microbial exopolysaccharides (EPS) on the heavy metal cadmium (Cd2+), this study took an EPS-producing Serratia fonticola CPSE11 (NZ_CP050171.1) isolated from Codonopsis pilosula root as the research object. The whole genome and EPS synthesis gene clusters of this strain were predicted and analyzed, the adsorption kinetics of EPS on Cd2+ were studied by using pseudo-first-order and second-order kinetic equations, the isothermal adsorption curves were simulated and analyzed by using the Langmuir isothermal adsorption equation, and the effects of Cd2+ and EPS on the growth of C. pilosula were explored by seed germination experiment and hydroponic experiment. The analysis revealed that this strain contained three gene clusters related to EPS synthesis, and the metabolic pathway for EPS synthesis was obtained on the basis of the whole genome analysis and microbial physiological metabolism. The molecular weight and monosaccharide composition of EPS were determined by HPLC analysis, which showed that EPS consisted of mannose, glucosamine, rhamnose, galactosamine, glucose, and galactose with a molar ratio of 1:1.74:4.57:3.96:14.04:10.28, with the molecular weight of 366,316.09 kDa. The adsorption process of EPS on Cd2+ was in accordance with the second-order kinetic model, and the results of seed germination experiments showed that EPS could promote seed germination and improve seed activity. In the hydroponic experiment, high concentration of Cd2+ (15 mg/L) caused toxic symptoms in C. pilosula, while the addition of EPS reduced the toxic effect of Cd2+ on C. pilosula, and the plant growth was significantly improved.

Genomic Characteristics and Comparative Genomics Analysis of the Endophytic Fungus Paraphoma chrysanthemicola DS-84 Isolated from Codonopsis pilosula Root.

Paraphoma chrysanthemicola is a newly identified endophytic fungus. The focus of most studies on P. chrysanthemicola has been on its isolation, identification and effects on plants. However, the limited genomic information is a barrier to further research. Therefore, in addition to studying the morphological and physiological characteristics of P. chrysanthemicola, we sequenced its genome and compared it with that of Paraphoma sp. The results showed that sucrose, peptone and calcium phosphate were suitable sources of carbon, nitrogen and phosphorus for this strain. The activities of amylase, cellulase, chitosanase, lipase and alkaline protease were also detected. Sequencing analysis revealed that the genome of P. chrysanthemicola was 44.1 Mb, with a scaffold N50 of 36.1 Mb and 37,077 protein-coding genes. Gene Ontology (GO) annotation showed that mannose-modified glycosylation was predominant in monosaccharide utilisation. The percentage of glycoside hydrolase (GH) modules was the highest in the carbohydrate-active enzymes database (CAZy) analysis. Secondary metabolite-associated gene cluster analysis identified melanin, dimethylcoprogen and phyllostictine A biosynthetic gene clusters (>60% similarity). The results indicated that P. chrysanthemicola had a mannose preference in monosaccharide utilisation and that melanin, dimethylcoprogen and phyllostictine A were important secondary metabolites for P. chrysanthemicola as an endophytic fungus.

Effects of time-space conversion on microflora structure, secondary metabolites composition and antioxidant capacity of Codonopsis pilosula root.

In order to study the relationship between medicinal plant Codonopsis pilosula phenotype, secondary metabolites, antioxidant capacity and its rhizosphere soil nutrients, root-related microorganisms under seasonal and geographical changes, high-throughput sequencing technology was used to explore the bacterial community structure and variation in rhizosphere soil and root endosphere from six regions of Dingxi City, Gansu Province during four seasons. Secondary metabolites composition and antioxidant capacities of C. pilosula root collected successively from four seasons were determined. The chemical properties, nutrient content and enzyme activities of rhizosphere of C. pilosula were significantly different under different temporal and spatial conditions. All soil samples were alkaline (pH 7.64-8.42), with water content ranging from 9.53% to 19.95%, and electrical conductivity varied widely, showing obvious time-scale effects. Different time scales were the main reasons for the diversity and structure of rhizosphere bacterial community of C. pilosula. The diversity and richness of rhizosphere bacterial community in autumn and winter were higher than those in spring and summer, and bacterial community structure in spring and summer was more similar to that in autumn and winter. The root length and diameter of C. pilosula showed significant time gradient difference under different spatiotemporal conditions. Nutrition and niche competition lead to significant synergistic or antagonistic interactions between rhizosphere bacteria and endophytic bacteria, which invisibly affect soil properties, abundance of functional bacteria and even yield and quality of C. pilosula. Soil properties, rhizosphere bacteria and endophytic bacteria directly promoted root phenotype, stress resistance and polysaccharide accumulation of C. pilosula.

Improved growth and metabolite accumulation in Codonopsis pilosula (Franch.) Nannf. by inoculation with the endophytic Geobacillu sp. RHBA19 and Pseudomonas fluorescens RHBA17.

In this study, we focused on the plant-growth-promoting properties of two strains isolated from Codonopsis pilosula, and the effect of inoculation with different strain treatments on physiological and metabolite accumulation of C. pilosula. The strains RHBA19 and RHBA17 were isolated and identified as Geobacillu sp. and Pseudomonas fluorescens, respectively. The two strains produced indole acetic acid (IAA), siderophore, biofilm, and various exoenzymes. Based on the pot experiments, inoculation of RHBA19 (G group) and the two mixed bacteria (M group) significantly improved the growth, root development, and photosynthesis of C. pilosula. Compared with the no-inoculation group (CK), the total polysaccharide content of root in the G and M groups was dramatically enhanced by 59.27% and 96.07%, and the lobetyolin (root) improved by 58.4% and 66.0%, respectively. After inoculation with bacteria agents, the activities of antioxidant enzymes (CAT, POD, SOD) of C. pilosula increased differentially. Inoculation with two types of bacterial agents significantly increased the activities of sucrose synthase (SS) and sucrose phosphate synthase (SPS) in root, and phenylalanine ammonia lyase (PAL) in leaf of C. pilosula. In addition, the content of signaling molecules (NO and H2O2) in three types of tissue increased significantly. The magnitude of these results was higher with mixtures than with individual strains. These results imply that the two types of bacterial agents induce physiological metabolism changes to accumulate polysaccharides and lobetyolin by regulating stress resistance enzymes and signal molecules, especially NO and H2O2.

Transporter drives the biosorption of heavy metals by Stenotrophomonas rhizophila JC1.

To better understand the function of transporter in heavy metal detoxification of bacteria, the transporters associated with heavy metal detoxification in S. rhizophila JC1 were analyzed, among which four members were verified by RT-qPCR. In addition, the removal rates of four single metal ions (Cr6+, Cu2+, Zn2+, Pb2+) and polymetallic ions by strain JC1 were studied, respectively. We also researched the physiological response of strain JC1 to different metal stress via morphological observation, elemental composition, functional group and membrane permeability analysis. The results showed that in the single metal ion solution, removal capacities of Cu2+ (120 mg/L) and Cr6+ (80 mg/L) of S. rhizophila JC1 reached to 79.9% and 89.3%, respectively, while in polymetallic ions solution, the removal capacity of each metal ion all decreased, and in detail, the adsorption capacity was determined Cr6+>Cu2+>Zn2+>Pb2+ under the same condition. The physiological response analyses results showed that extracellular adsorption phenomena occurred, and the change of membrane permeability hindered the uptake of metal ions by bacteria. The analysis of transporters in strain JC1 genome illustrated that a total of 323 transporters were predicted. Among them, two, six and five proteins of the cation diffusion facilitator, resistance-nodulation-division efflux and P-type ATPase families were, respectively, predicted. The expression of corresponding genes showed that the synergistic action of correlative transporters played important roles in the process of adsorption. The comparative genomics analysis revealed that S. rhizophila JC1 has long-distance evolutionary relationships with other strains, but the efflux system of S. rhizophila JC1 contained the same types of metal transporters as other metal-resistant bacteria.

Comparative analysis of three kinds of extraction kinetic models of crude polysaccharides from Codonopsis pilosula and evaluate the characteristics of crude polysaccharides.

In this study, the second-order model, Fick's second law of diffusion, and the Peleg model were used to evaluate the extraction kinetic model of polysaccharide (CPP) from Codonopsis pilosula. The characteristic functional groups, surface structure, and physical and chemical properties of CPP were analyzed by multi-spectroscopic and microscopic techniques. The results showed that the extraction process agreed well with the second-order model, Fick's second diffusion law, and Peleg model. Rheological tests showed that CPP exhibited different viscosity changes under different conditions (Solution viscosity was inversely proportional to temperature, time, etc.; proportional to polysaccharide concentration, Na+ content, etc.). CPP was composed of molecular aggregates composed of small particles, with more pore structure and basically completely decomposed at 130 °C. The hypoglycemic study showed that CPP had a strong inhibitory effect on α-glycosidase than α-amylase. The morphology and subsequent structural features, anti-diabetic potential, and rheological properties of CPP were revealed to provide a theoretical basis for the development of pharmaceutical preparations or health food and functional food for the treatment of diabetes. Supplementary Information: The online version contains supplementary material available at 10.1007/s13399-022-02518-w.

Extraction kinetic model of polysaccharide fromCodonopsis pilosulaand the application of polysaccharide in wound healing.

The crude polysaccharide (CPNP) ofCodonopsis pilosulawas obtained by hot-water extraction technology. The extraction kinetic model established according to Fick's first law of diffusion and related parameters of polysaccharide was studied. CPNP microcapsules were prepared by blending with sodium alginate, Ca2+ions and crude CPNP. The quality control (drug loading rate, embedding rate and release rate, etc) of CPNP microcapsules were analyzed by pharmacopeas standards. The structure feature of CPNP microcapsules also were determined with various methods. The wound healing ability of CPNP microcapsules loading with different concentration of CPNP was evaluated using the rat wound model. The activity of various enzymes and the expression levels of pro-inflammatory factors in the model skin tissue also were determined by enzyme linked immunosorbent assay (ELISA). Hematoxylin-eosin staining (HE), Masson, immunohistochemistry were used to investigate the external application effect of CPNP microcapsules on skin wound repair. The extraction kinetics of CPNP was established with the linear correlation coefficient (R2) of 0.83-0.93, implied that the extraction process was fitted well with the Fick's first law of diffusion. The CPNP has good compatibility with sodium alginate and Ca2+ions by SEM and TEM observation, and the particle size of CPNP microcapsules was 21.25 ± 2.84 µm with the good degradation rate, loading rate (61.59%) and encapsulation rate (55.99%), maximum swelling rate (397.380 ± 25.321%). Compared with control group, the redness, and swelling, bleeding, infection, and exudate of the damaged skin decreased significantly after CPNP microcapsules treatment, and the CPNP microcapsules groups exhibited good wound healing function with less inflammatory cell infiltration. The pathological structure showed that in the CPNP microcapsules group, more newborn capillaries, complete skin structure, and relatively tight and orderly arrangement of collagen fibers were observed in the skin of rats. CPNP microcapsules could effectively inhibit the high expression of pro-inflammatory factors in damaged skin, and significantly increase the contents of related enzymes (GSH-Px, T-AOC, LPO) and collagen fibers. The relative expression levels of genes (VEGF and miRNA21) in the CPNP microcapsules group were higher than those in the model group and the negative group. The above results suggested that the CPNP microcapsules could controlled-release the CPNP to the wound surface, and then played a better role in antibacterial, anti-inflammatory and skin wound repair.