Effect of Chitosan and Hyperbranched Poly-L-Lysine Treatment on Quality of Cucumber (Cucumis sativus L.) during Storage.

To enhance the storage time of cucumbers, this research investigated the impact of chitosan (CS) and hyperbranched poly-L-lysine (HBPL) on the quality and nutritional attributes of cucumbers when stored at a temperature of 25 °C. The results demonstrated that sensory evaluation scores for cucumbers treated with a CS-HBPL combination were significantly higher than the control (CK), CS, and HBPL groups. On the 18th day of storage, cucumbers in the CK group exhibited significant decay and softening; however, there was a decrease in hardness observed in the CS-HBPL group and no decay or noticeable sour taste was detected. Furthermore, compared to the CK group, treatment with CS-HBPL effectively delayed cucumber decay and weight loss rate while significantly inhibiting decreases in cucumber hardness and growth of surface microorganisms. Additionally, it substantially reduced losses of soluble protein content as well as vitamin C (Vc), reducing sugars, and total phenolic compounds within cucumbers, which were 4.7 mg/g, 4.7 mg/g, 0.94 mg/g, and 0.52 mg/kg, respectively. Moreover, compared to the CK group, combined treatment with CS-HBPL significantly inhibited malondialdehyde (MDA) accumulation and reducing relative electrolyte permeability within cucumbers, which were 1.45 µmol·g-1FW and 29.82%. Furthermore, it notably enhanced activities of superoxide dismutase (SOD) and catalase (CAT), while exerting a significant inhibitory effect on polyphenol oxidase (PPO). In summary, the combined CS-HBPL treatment successfully prolonged cucumber shelf life at room temperature, enabling new possibilities for extending cucumber shelf life.

Single-cell and spatial RNA sequencing reveal the spatiotemporal trajectories of fruit senescence.

The senescence of fruit is a complex physiological process, with various cell types within the pericarp, making it highly challenging to elucidate their individual roles in fruit senescence. In this study, a single-cell expression atlas of the pericarp of pitaya (Hylocereus undatus) is constructed, revealing exocarp and mesocarp cells undergoing the most significant changes during the fruit senescence process. Pseudotime analysis establishes cellular differentiation and gene expression trajectories during senescence. Early-stage oxidative stress imbalance is followed by the activation of resistance in exocarp cells, subsequently senescence-associated proteins accumulate in the mesocarp cells at late-stage senescence. The central role of the early response factor HuCMB1 is unveiled in the senescence regulatory network. This study provides a spatiotemporal perspective for a deeper understanding of the dynamic senescence process in plants.

A novel GARP humanized mouse model for efficacy assessment of GARP-targeting therapies.

Although breakthroughs have been achieved with immune checkpoint inhibitors (ICI) therapy, some tumors do not respond to those therapies due to primary or acquired resistance. GARP, a type I transmembrane cell surface docking receptor mediating latent transforming growth factor-ß (TGF-ß) and abundantly expressed on regulatory T lymphocytes and platelets, is a potential target to render these tumors responsive to ICI therapy, and enhancing anti-tumor response especially combined with ICI. To facilitate these research efforts, we developed humanized mouse models expressing humanized GARP (hGARP) instead of their mouse counterparts, enabling in vivo assessment of GARP-targeting agents. We created GARP-humanized mice by replacing the mouse Garp gene with its human homolog. Then, comprehensive experiments, including expression analysis, immunophenotyping, functional assessments, and pharmacologic assays, were performed to characterize the mouse model accurately. The Tregs and platelets in the B-hGARP mice (The letter B is the first letter of the company's English name, Biocytogen.) expressed human GARP, without expression of mouse GARP. Similar T, B, NK, DCs, monocytes and macrophages frequencies were identified in the spleen and blood of B-hGARP and WT mice, indicating that the humanization of GARP did not change the distribution of immune cell in these compartments. When combined with anti-PD-1, monoclonal antibodies (mAbs) against GARP/TGF-ß1 complexes demonstrated enhanced in vivo anti-tumor activity compared to monotherapy with either agent. The novel hGARP model serves as a valuable tool for evaluating human GARP-targeting antibodies in immuno-oncology, which may enable preclinical studies to assess and validate new therapeutics targeting GARP. Furthermore, intercrosses of this model with ICI humanized models could facilitate the evaluation of combination therapies.

Mapping the immunological battlefield in gastric cancer: prognostic implications of an immune gene expression signature.

BACKGROUND: Gastric cancer (GC) is a heterogeneous malignancy with variable clinical outcomes. The immune system has been implicated in GC development and progression, highlighting the importance of immune-related gene expression patterns and their prognostic significance. OBJECTIVE: This study aimed to identify differentially expressed immune-related genes (DEIRGs) and establish a prognostic index for GC patients using comprehensive bioinformatic analyses. METHODS: We integrated RNA sequencing data from multiple databases and identified DEIRGs by overlapping differentially expressed genes with immune-related genes. Functional enrichment analysis was performed to uncover the biological processes and signaling pathways associated with DEIRGs. We conducted a Weighted Gene Co-expression Network Analysis (WGCNA) to identify key gene modules related to with GC. Cox regression analysis was conducted to determine independent prognostic DEIRGs for overall survival prediction. Based on these findings, we developed an immune-related gene prognostic index (IRGPI) based on these findings. The prognostic value of the IRGPI was validated using survival analysis and an independent validation cohort. Functional enrichment analysis, gene mutation analysis, and immune cell profiling were performed to gain insights into the biological functions and immune characteristics associated with the IRGPI-based subgroups. RESULTS: We identified 493 DEIRGs significantly enriched in immune-related biological processes and signaling pathways associated with GC. WGCNA analysis revealed a significant module (turquoise module) associated with GC, revealing potential therapeutic targets. Cox regression analysis identified RNASE2, CGB5, CTLA4, and DUSP1 as independent prognostic DEIRGs. The IRGPI, incorporating the expression levels of these genes, demonstrated significant prognostic value in predicting overall survival. The IRGPI-based subgroups exhibited distinct biological functions, genetic alterations, and immune cell compositions. CONCLUSION: Our study identified DEIRGs and established a prognostic index (IRGPI) for GC patients. The IRGPI exhibited promising prognostic potential and provided insights into GC tumor biology and immune characteristics. These findings have implications for guiding therapeutic strategies.

UV-ARTP-DES compound mutagenesis breeding improves natamycin production of Streptomyces natalensis HW-2 and reveals transcriptional changes by RNA-seq.

Natamycin is widely used in food, medical and health, agriculture, and animal husbandry. In this study, Streptomyces natalensis HW-2 was used as the research object, and a mutant DES-26 with stable genetic characters was selected by UV-ARTP-DES compound mutation. The natamycin yield was 1.64 g/L, 86.36% higher than original strain. Differential expression genes were analyzed by transcriptomics, and results showed that 295 and 860 genes were significantly differentially expressed at fermentation for 48 h and 72 h. GO and KEGG analysis showed that compound mutagenesis had a significant impact on glycolysis, pentose phosphate, TCA cycle, fatty acid metabolism pathways, and several key enzyme genes in the pathways were up-regulated, and genes related to natamycin biosynthesis (pimB-pimI) and transcriptional regulator (pimR) were also up-regulated. qRT-PCR results confirmed that expression levels of these genes were consistent with transcriptional changes of RNA-Seq. Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-022-01191-z.

CsMYC2 is involved in the regulation of phenylpropanoid biosynthesis induced by trypsin in cucumber (Cucumis sativus) during storage.

Trypsin has a new activity of scavenging superoxide anion and generating hydrogen peroxide. Trypsin can significantly improve the storage quality of C. sativus. To illustrate the mechanism of trypsin-induced resistance in fruits and vegetables, an integrated analysis of widely targeted metabolomics and transcriptomics was carried out. Transcriptomic results showed that 1068 genes highly related to phenylpropanoid biosynthesis gathered in the brown module were obtained by WGCNA. In KEGG analysis, differentially expressed genes (DEGs) were also highly enriched in EIP (Environmental Information Processing) pathways "Plant hormone signal transduction (map04075)" and "MAPK signaling pathway-plant (map04016)". Next, 87 genes were identified as the leading edge by GSEA analysis. So far, CsMYC2 was highlighted as a key transcription factor that regulates phenylpropanoid biosynthesis identified by GSEA and WGCNA. Furthermore, the major route of biosynthesis of phenylpropanoid compounds including coumarins, lignins, chlorogenic acid, flavonoids, and derivatives regulated by trypsin was also illustrated by both transcriptomic and metabolomic data. Results of O2PLS showed that CsMYC2 was positively correlated with Rosmarinic acid-3-O-glucoside, Epigallocatechin, Quercetin-3-O-sophoroside (Baimaside), and so on. Correlation between CsMYC2, phenylpropanoid related genes, and metabolites in C. sativus was illustrated by co-expression networks. Roles of CsMYC2 were further checked in C. sativus by VIGS. The results of this study might give new insight into the exploration of the postharvest resistance mechanism of C. sativus induced by trypsin and provide useful information for the subsequent mining of resistance genes in C. sativus.

Effect of Combined Chitosan and Hyperbranched Poly-L-Lysine Based Coating on Prolonging the Shelf Life of Oyster Mushroom (Pleurotus ostreatus).

To extend the shelf life of oyster mushroom (Pleurotus ostreatus), the effects of chitosan (CS) and hyperbranched poly-L-lysine (HBPL) combined treatment on quality characteristics, nutritional quality, storage characteristics, and enzyme activity of oyster mushroom during postharvest storage at 4 °C were investigated. The results showed that CS-HBPL combined treatment could significantly reduce rot degree and weight loss and significantly inhibit the browning of oyster mushroom. At the same time, the loss of reducing sugar, vitamin C, soluble protein, and total phenolic was significantly reduced. Compared with the control, CS-HBPL combined treatment could also significantly inhibit an increase in malondialdehyde (MDA) and significantly decrease the relative electrolyte leakage of oyster mushroom. In addition, the activities of catalase (CAT), superoxide dismutase (SOD), phenylalnine ammonialyase (PAL), and peroxidase (POD) were significantly improved, and the activity of polyphenol oxidase (PPO) was significantly inhibited in oyster mushroom. In conclusion, CS-HBPL combined treatment had a good protective effect on the membrane permeability damage of oyster mushroom and could effectively delay the oxidation of phenolic substances and browning of oyster mushroom. Therefore, CS-HBPL combined treatment can be used as a potential strategy to extend the storage time of oyster mushroom.

Transcriptome analyses and virus-induced gene silencing identify HuWRKY40 acting as a hub transcription factor in the preservation of Hylocereus undatus by trypsin.

Trypsin can significantly improve the storage quality of Hylocereus undatus (H. undatus). To verify the hub WRKY gene of H. undatus in trypsin preservation, joint analysis of transcriptome and protein-protein interaction (PPI) network was carried out, and virus-induced gene silencing (VIGS) was conducted. In the transcriptome of H. undatus, GO directed acyclic graph (DAG) showed that the GO terms of 55 WRKY genes were mainly enriched in sequence-specific DNA binding, DNA binding transcription factor activity, and so on. The GO enrichment chord diagram showed that HuWRKY40 was significantly up-regulated in the enriched top10 GO terms. KEGG enrichment analysis showed that 55 WRKY genes were mainly enriched in plant-pathogen interaction and MAPK pathway. The results of PPI network showed that HuWRKY40 was a hub protein of WRKY transcription factors (TFs) family regulated by trypsin, which was consistent with the results of transcriptome analysis. Bioinformatics analysis showed that HuWRKY40 of H. undatus had the highest homology with Beta vulgaris L. and Spinacia oleracea L. The function of the core regulatory protein HuWRKY40 was further clarified by VIGS technology. The results of VIGS showed that there was a big difference between the phenotype of the pTRV2-HuWRKY40 group and that of the control group. Finally, it was confirmed that HuWRKY40 accelerated the synthesis of flavonoids and improved the fruit quality during the storage of H. undatus. This study found that trypsin may regulate HuWRKY40 activity through the MAPK cascade pathway, affect the participation of flavonoid synthesis, and then delay fruit corruption. PRACTICAL APPLICATIONS: With attention of people to the safety and freshness of fruits and vegetables, biological preservation technology has become one of the hotspots in the field of preservation in recent years. Trypsin can significantly improve the antioxidant capacity of fruits and vegetables. As a new biological preservative, it is convenient to operate and economical. In the current work, the mechanism of trypsin on the WRKY TFs during H. undatus storage was investigated. The application of trypsin would provide a new strategy for the storage quality control of fruits and vegetables.

Microwave Irradiation: Effects on the Change of Colour Characteristics and Main Phenolic Compounds of Cabernet Gernischt Dry Red Wine during Storage.

Wine colour is an essential organoleptic property considered by consumers. In this paper, the potential effects on colour characteristics and the content of main phenolic compounds in red wine under microwave irradiation were investigated during wine storage. The results showed that the changing trend of colour characteristics of microwave-treated and untreated wines was very similar. Moreover, total phenolic compounds, total monomeric anthocyanins, main anthocyanins, main flavonoids, and main phenolic acids (gallic acid; caffeic acid; syringic acid; (+)-catechin; Cy-3-glu; Mv-3-glu) also showed similar change trends during storage. In other words, microwave irradiation had a long-term effect on the colour properties and main phenolic compounds of red wine, changes that require long-time aging in traditional processing. In terms of the studied parameters, the changes in microwave-treated wine were faster than those in untreated wine. These results showed that microwave technology, as a promising artificial aging technology, could in a short time produce red wine of similar quality to traditional aging.

Biotechnological production and application of epsilon-poly-L-lysine (ε-PL): biosynthesis and its metabolic regulation.

Epsilon-poly-L-lysine (ε-PL) is an unusual biopolymer composed of L-lysine produced by several microorganisms, especially by the genus Streptomyces. Due to its excellent antimicrobial activity, good water solubility, high safety, and biodegradable nature, ε-PL with a GRAS status has been widely used in food and pharmaceutical industries. In the past years, studies have focused on the biotechnological production of ɛ-PL, the biosynthetic mechanism of microbial ɛ-PL, and its application. To provide new perspectives from recent advances, the review introduced the methods for the isolation of ɛ-PL producing strains and the biosynthetic mechanism of microbial ɛ-PL. We summarized the strategies for the improvement of ɛ-PL producing strains, including physical and chemical mutagenesis, ribosome engineering and gene engineering, and compared the different metabolic regulation strategies for improving ɛ-PL production, including medium optimization, nutrient supply, pH control, and dissolved oxygen control. Then, the downstream purification methods of ɛ-PL and its recent applications in food and medicine industries were introduced. Finally, we also proposed the potential challenges and the perspectives for the production of ε-PL.

Effects of ethanol stress on epsilon-poly-l-lysine (ε-PL) biosynthesis in Streptomyces albulus X-18.

The aim of the study was to reveal the effects of ethanol stress on the production of epsilon-poly-l-lysine (ε-PL) in Streptomyces albulus X-18. The results showed that biomass and the utilization of glucose were respectively increased by ethanol stress. The ε-PL yield was increased by 41.42 % in the shake flask and 37.02 % in 10 L fermenter with 1% (v/v) ethanol. The morphology of colonies and mycelia showed significant differences. The intracellular reactive oxygen species level was increased by about 100 %. The ratio of unsaturated fatty acids to saturated fatty acids in the cell membrane was increased by ethanol stress. Isobaric Tags for Relative and Absolute Quantitation (iTRAQ) proteomic profile showed that 265 identified proteins were differentially expressed. The differentially expressed proteins (DEPs) were mainly involved in biological processes. The up-regulated DEPs were mainly involved in the redox reaction and stress response. The metabolic flux of l-Asp was shifted to l-Lys biosynthesis, and the DAP pathway was strengthened. Protein-protein interaction analysis showed that 30 DEPs interacted with l-Lys biosynthesis. The changes of ten proteins by Parallel Reaction Monitoring (PRM) were consistent with those by iTRAQ. The study provided valuable clues to better understand the mechanism of ε-PL biosynthesis improvement by ethanol stress.

Amplified detection signal at a photoelectrochemical aptasensor with a poly(diphenylbutadiene)-BiOBr heterojunction and Au-modified CeO2 octahedrons.

A major aspect of this work is the synergistic application of a poly(diphenylbutadiene)-BiOBr composite and a gold nanoparticle-linked CeO2 octahedron to develop a photoelectrochemical aptasensor with an easily measurable detection signal change. Specifically, poly(diphenylbutadiene) nanofiber-immobilised BiOBr flower-like microspheres were developed as a hybrid material with a heterojunction that facilitates high visible light absorption and efficient photo-generated charge separation, which are essential features for sensitive photoelectrochemical sensors. The model analyte acetamiprid was attached via its specific aptamer on the aptasensor. Separately, a gold nanoparticle-linked CeO2 octahedron was strategically used to significantly diminish the photocurrent by impeding electron transfer at the aptasensor surface. After acetamiprid binding, the CeO2 octahedrons were displaced from the aptasensor. This caused a weakened quenching effect and restored the photocurrent to accomplish an "on-off-on" detection mechanism. This photoelectrochemical aptasensor exhibited a detection limit of 0.05 pM over a linear range of 0.1 pM-10 µM acetamiprid. The use of an aptamer has provided good specificity to acetamiprid and anti-interference. In addition, an ∼5.8% relative standard deviation was estimated as the reproducibility of the photoelectrochemical aptasensor. Furthermore, nearly 90% of the initial photocurrent was still measurable after storing these aptasensors at room temperature for 4 weeks, demonstrating their stability.

[Advances in the biosynthesis of natamycin and its regulatory mechanisms].

Natamycin is a polyene macrolide antibiotics with strong and broad spectrum antifungal activity. It not only effectively inhibits the growth and reproduction of fungi, but also prevents the formation of some mycotoxins. Consequently, it has been approved for use as an antifungal food preservative in most countries, and is also widely used in agriculture and healthcare. Streptomyces natalensis and Streptomyces chatanoogensis are the main producers of natamycin. This review summarizes the biosynthesis and regulatory mechanism of natamycin, as well as the strategies for improving natamycin production. Moreover, the future perspectives on natamycin research are discussed.

Analysis of chemical compounds and toxicological evaluation of Forsythia suspensa leaves tea.

To determine the compositions of Forsythia suspensa leaves tea (FSLT) and its safety, the chemical compounds were analysed with some methods, and the toxicity was evaluated in Kunming mice and Wistar rats. The results showed that FSLT contained rich flavonoid, lignans, triperpene acids, amino acids, and mineral elements. In the acute toxicity study, none of the mice died, and no obvious poisoning symptoms were observed after 14 days in mice at the dose of 15 mg/g·body weight (bw) FSLT; in the sub-chronic toxicity, no abnormal or dead rat was found at the dose of 1, 3, and 10 mg/g·bw during 90 days feeding administration; there was no significant difference in bw and food consumption; no significant differences were found in each hematology and serum biochemistry parameter and organ/body weight ratio comparing with the control experimental group. The results revealed that the FSLT has low or no toxicity via oral administration. Therefore, FSLT is very suitable and safe to be used as a new resource food.

Rhizoglomus intraradices Improves Plant Growth, Root Morphology and Phytohormone Balance of Robinia pseudoacacia in Arsenic-Contaminated Soils.

Arbuscular mycorrhizal fungi (AMF) are known to improve the resistance of host plants against various heavy metal stresses. However, the arsenic (As) resistance mechanism of AMF-inoculated woody legumes remains unclear. In this study, black locust (Robinia pseudoacacia L.) seedlings were cultivated in potted soils inoculated with or without AMF Rhizoglomus intraradices under three different levels of As stress (0, 100, and 200 mg As kg-1 soil) over 4 months. The objective of this paper was to investigate the effects of AMF on plant growth, root morphology, and the content and ratio of endogenous phytohormones and soil glomalin under As stress condition. As stress toxicity suppressed the AM spore germination and colonization, plant growth, and the content of soil glomalin and changed the morphological characteristics of the roots and the balance of endogenous hormone levels in plants. However, R. intraradices inoculation improved the shoot and root dry weights, total root length, root surface area, root volume, and the number of root forks and tips across all As treatments. R. intraradices inoculation obviously decreased the percentage of root length in the 0- to 0.2-mm diameter class and increased those in the 0.5- to 1.0-mm and >1.0-mm diameter classes; the percentages in the 0.2- to 0.5-mm diameter class were less affected by R. intraradices inoculation. The concentrations of the easily extractable glomalin-related (EE-GRSP) and total glomalin-related soil protein (T-GRSP) were higher in the of R. intraradices-inoculated seedlings than those in the non-inoculated seedlings. Furthermore, R. intraradices inoculation increased the concentrations of indole-3-acetic acid (IAA) and abscisic acid (ABA), but decreased the concentrations of gibberellic acid (GA) and zeatin riboside (ZR). The phytohormone ratios of IAA/ABA, GA/ABA, ZR/ABA, and (IAA + GA + ZR)/IAA in the R. intraradices-inoculated seedlings were lower than those in the non-inoculated seedlings. These results indicated that R. intraradices alleviated As toxicity in R. pseudoacacia seedlings by improving their plant growth, altering root morphology, regulating the concentrations and ratios of phytohormones, and increasing the concentration of soil glomalin. The results suggested that AMF-inoculated R. pseudoacacia seedlings would be a critical factor in successful vegetation restoration and soil development in As-contaminated soils.

Screening of cardioprotective diseases bioactive components from Danshen extracts and LC-MS analysis.

A rapid and efficient analysis and screening method is adopted for cell affinity capture coupled with HPLC-MS (CAC-HPLC-MS) analysis of bioactive components that have possible efficiency against cardiovascular diseases. This method involves affinity capture, concentration, and separation of bioactive components from Danshen library using oxidatively damaged endothelial cells induced by H2 O2 , as well as analysis and identification of targeted compounds with HPLC and MS. It combines the specific interaction between cell membrane receptors and bioactive components with the powerful analysis and identification function of HPLC-MS. The CAC-HPLC-MS method was also used for analysis and screening of bioactive components from crude extracts of Danshen. A total of 19 components were found to be bound to oxidatively damaged endothelial cells with seven of these identified. Existing literature confirms that these seven components have many activities related to cardioprotective diseases. Therefore, the combination of biological affinity capture with HPLC-MS should be regarded as an attractive method with great potential for rapid and efficient screening of bioactive components related to anti-cardiovascular diseases from natural product libraries.

Protein-protein interaction site predictions with minimum covariance determinant and Mahalanobis distance.

Protein-protein interaction site (PPIS) prediction must deal with the diversity of interaction sites that limits their prediction accuracy. Use of proteins with unknown or unidentified interactions can also lead to missing interfaces. Such data errors are often brought into the training dataset. In response to these two problems, we used the minimum covariance determinant (MCD) method to refine the training data to build a predictor with better performance, utilizing its ability of removing outliers. In order to predict test data in practice, a method based on Mahalanobis distance was devised to select proper test data as input for the predictor. With leave-one-validation and independent test, after the Mahalanobis distance screening, our method achieved higher performance according to Matthews correlation coefficient (MCC), although only a part of test data could be predicted. These results indicate that data refinement is an efficient approach to improve protein-protein interaction site prediction. By further optimizing our method, it is hopeful to develop predictors of better performance and wide range of application.

Identification of a Novel Antibiotic from Myxobacterium Stigmatella Eracta WXNXJ-B and Evaluation of its Antitumor Effects I n - vitro.

This work was to isolate and identify the bioactive secondary metabolite which was produced by myxobacterium Stigmatella eracta WXNXJ-B, and to evaluate its antitumor and apoptosis-inducing effects. The results showed that one novel compound (molecular formula C29H25NO3) was isolated, purified by Sephadex LH-20 column chromatography and preparative RP-HPLC, and identified as 5-(6-benzyl-quinolin-3-ylmethyl)-6- phenyl-3,7-dioxa- bicycle [4.1.0] heptan-3-one (named as quinoxalone) according to its UV, IR, HRMS and NMR spectra. The compound showed strong antitumor activity on B16, HepG2, MCF-7, SGC-7901, MDA-MB231 and CT-26 six tumor cell lines in-vitro. Nevertheless, it showed less cytotoxic to the mouse normal spleen cells (IC50 was 836.27 ± 13.02 µg mL(-1)). The cytotoxic study on HepG2 cells in-vitro showed that quinoxalone could induce the change of cell nuclear and arrested the cell division in the S and G2/M phase. Our results suggest that quinoxalone could be a potential anti-cancer agent.

Influence of fungal elicitors on biosynthesis of natamycin by Streptomyces natalensis HW-2.

To investigate the effect of fungal elicitors on biosynthesis of natamycin in the cultures of Streptomyces natalensis HW-2, the biomass and filtrate of the broth from Aspergillus niger AS 3.6472, Penicillium chrysogenum AS 3.5163, A. oryzae AS 3.2068, and Saccharomyces cerevisiae AS 2.2081 were used to induce natamycin production in S. natalensis HW-2. The results showed that the biomass of P. chrysogenum AS 3.5163 could enhance the yield of natamycin from 0.639 to 0.875 g l(-1). The elicitor from the fermentation broth of P. chrysogenum AS 3.5163 showed the highest inducing efficiency with the yield of natamycin enhanced from 0.632 to 1.84 g l(-1). The elicitor that was cultured for 2 days showed the strongest inducing activity during the fermentation of S. natalensis HW-2 for 24 h, and the yield of natamycin was enhanced from 0.637 to 2.12 g l(-1). The biochemical parameters were examined at the end of fermentation and the results demonstrated that both the growth of cells and the concentration of residual sugar could be influenced. The residual sugar decreased from 5.03 to 4.27 g l(-1), and the biomass decreased from 10.26 to 6.87 g l(-1). Finally, the elicitor was identified as a low molecular weight substance with a similar polarity to that of butyl alcohol by primary qualitative analysis.

Dialysis sampling on-line coupled with high-performance liquid chromatography for simultaneous investigation of the interactions between multi-components in herbs and the albumin.

An on-line dialysis sampling method coupled with high-performance liquid chromatography was developed for simultaneous investigation of the interactions between multiple bioactive compounds contained in herbal medicines and proteins. The system was used to estimate the interactions of components in danshen (Salvia miltiorrhiza) injection with bovine serum albumin. The results showed that the binding actions of five water-soluble compounds in danshen injection could be simultaneously investigated. The binding parameters of caffeic acid, ferulic acid, danshensu, protocatechuic acid and protocatechuic aldehyde were obtained and the interaction mechanisms were explored. The association constant evaluated for caffeic acid agreed well with literature values. The proposed approach should be beneficial for examining the holistic combined action of herbal medicine with proteins and help facilitate the discovery process of drug candidates.