Papers by Author: Jin Ju Ding

Abstract: Purpose To prepare Grifola frondosa fermented beverage and study on its production process, ingredients and characteristics. Methods The following process was used to prepare Grifola frondosa fermented beverage: Grifola frondosa original strains →Strain activation →Inclined plane strain →Liquid spawn →Submerged fermentation →Fermentation broth with mycelium →Colloid mill grinding →High pressure homogenization →Homogenate →Add preservatives or not →Package → Sterilizing or not →Finished product. Grifola frondosa mycelium biomass was determined by Weight-Drying method. Total sugar content was measured by Phenol-sulfuric acid method. Reducing sugar content was measured by 3, 5-dinitrosalicylic acid (DNS) method. Protein content was measured by Coomassie brilliant blue method. The quality of Grifola frondosa fermented beverage was evaluated every 10 days from six areas: color, precipitation, transparency, aroma, taste and the bacteria growth situation. Results Mycelia biomass of Grifola frondosa fermentation product is 378.3 ± 7.21mg/100ml. The total sugar content is 1.8995 mg/ml, the reducing sugar content is 0.2539 mg/ml, and grifolan content is 1.6456 mg/ml, protein content 7.971μg/ml. The total number of colonies is 67 CFU/ml and no E. coli in Grifola frondosa fermented beverage, this comply with the requirements of the national food safety standard. The samples of Grifola frondosa fermented beverage without sterilized and preservatives placed at room temperature for 6 months are normal, and no bacteria growth. Conclusion The production process of Grifola frondosa fermented beverage is feasible. In the case without sterilization and preservatives, Grifola frondosa fermented beverage can be placed at room temperature for 6 months without deterioration and still maintain its characteristics of nutrient-rich, pure taste, clean and hygienic, with natural mushroom fragrant. This can provide a reference for the development of Grifola frondosa submerged fermentation products.

Abstract: Purpose To search for a purification method of Grifolan Synthase crude extract from the mycelium of Grifola frondosa and determine its molecular weight. Methods Grifolan Synthase crude extract from the mycelium of Grifola frondosa was purified by recrystallization and native-PAGE. The process is as follows: Grifolan Synthase crude extract was dissolved in buffer solution (pH7.0). 10ml Grifolan Synthase crude extract solution was mixed with ammonium sulfate slowly to the concentration of 60%, and then mixed with 3ml cooling acetone (refrigeration at -18 °C for 12 hours) slowly, after standing for 24 hours, centrifugated at 5000rpm for 10 minutes at 4°C. Material at the interface was collected and air-dried as higher purity Grifolan Synthase. The higher purity Grifolan Synthase was dispersed in the native-PAGE gel, and the active band of native-PAGE gel was cut down and broken by ultrasonic treatment for 1min, then centrifugated at 5000rpm for 10 minutes at 4°C, the supernatant was taken and mixed with ammonium sulfate slowly to the concentration of 60%, after 24 hours on standing, centrifugated at 12000rpm for 10 minutes at 4°C, the pellets was got and the purity was checked by SDS-PAGE. The molecular weight of Grifolan Synthase was determined by SDS polyacrylamide gel electrophoresis (SDS-PAGE). Results Grifolan Synthase purified by recrystallization and native-PAGE was checked to be a single band by SDS-PAGE. The data of molecular weight obtained by SDS-PAGE showed that the molecular weight of Grifolan Synthase was 55000Da. Conclusion A purification method of Grifolan Synthase crude extract from the mycelium of Grifola frondosa was researched out, and the molecular weight of grifolan synthase was studied in this paper. It can lay the foundation for the further study on the structure and function of Grifolan Synthase.

Abstract: Purpose To develop a novel complex antimicrobial agent and determine the optimal components of the composite antimicrobial agents and its antimicrobial activity in vitro. Methods According to antimicrobial mechanisms，antibacterial spectrums，physical and chemical properties and applicabilities of existing antimicrobial agents in clinical use, select out cefoperazone sodium, sulbactam sodium and cephradine as the basic components to make a novel complex antimicrobial agent. Utilize yeast, staphylococcus aureus and E. coli bacteria as test bacteria. Do the three factors four-level orthogonal experiments by the maximum amount, the middle amount, low amount and Minimum amount of the three-component agent to research the optimum ratio of the drug. Measure the titer of the compound antimicrobial agent by the way of tube-plate method (2 doses). With known contents of Penicillin Sodium for Injection as control, and determine its minimum inhibitory concentration against staphylococcus aureus, E. coli and yeast by using the agar doubling dilution method. The experimental results were analysized by statistical analysis software SPSS16.0. Results The results of the three factors four-level orthogonal experiments indicate the optimum ratios of Cefoperazone Sodium, Sulbactam Sodium and Cephradine against E. coli, yeast and staphylococcus aureus were 2:2:3, 1:2:2 and 2:6:5, their titers were 1353.9U/mg, 982.7U/mg and 1015.5U/mg. With the highest titer proportion 2:2:3 as the composition of the antimicrobial compound. This compound antimicrobial agent had a good antimicrobial activity against Gram-positive bacteria, Gram-negative bacteria and Fungi, its minimal inhibitory concentration (MIC) against staphylococcus aureus, E. coli and yeast were 2.000μg/ml, 0.500μg/ml and 16.000μg/ml. Conclusion This research acquires a composite of antibiotics. This antimicrobial compound has a broader spectrum and higher antimicrobial activity in vitro comparing with traditional common single antibiotics, and it especially has a good antimicrobial activity against fungi. The results set a scientific foundation for enriching clinical medicines.

Abstract: To prepare out a novel preparation of liuweidihuang which effectively preserves Paeonol, Ursolic acid and Polysaccharides, and the preparation process should be scientific, lower-price and simple. The microscopic characteristics and its physicochemical properties of super-micro-particle of liuweidihuang from above process were also discussed. Methods According to the therapeutic and physicochemical properties of materials in liuweidihuang, the processed technology is as follow: Ripe rehmannia glutinosa,Tuckahoe and Alisma digitalis were boiled for 4.5h., the root bark of the peony tree(Paeonia suffruticosa), dogberry and yam were smashed to nanoparticles. Then select the reasonable method to drying the mixture. The microscopic shape characteristics of the before- and after- broken particles was compared. Paeonol content and dissolution were determined by HPLC. The stability and fluidity of liuweidihuang nanoparticles were examined by precipitation and funnel way. Results The approach of prepared liuweidihuang nano-microcapsules is scientific, lower-price and simple. The average size of processed super-micro-particle of liuweidihuang is 400±46nm. The hardness of the table is 9.7kg/cm2. The thickness of the table is 0.6cm. The friability of the tablet is ＜1%. Each tested item complied with standards of troches of Chinese medicine, and it is beneficial to industrial production for Chinese medicine.. Conclusion The nano-microcapsule is dispersive, desiccative, mobilizable, stable and three times concentrated than traditional pills of Liuweidihuang. It also can be used to produce different forms of drugs in the market.

Abstract: Objective In order to increase the Paeonol dissolution and content, cortex moutan were smashed into nanoparticles, and the dissolution and content were compared by microscopy before and after super-micro-particle pulverization. Methods Super-micro-particle pulverization and general grinding were used to broke Cortex moutan into particles. The microscopic morphous characteristics of the before- and after- ultra-disintegration particles were compared by microscopy. Methods of HPLC was used to determine the content and dissolution of Paeonol with different grinding conditions. Methods of precipitation and funnel way were used to examine the stability and fluidity of cortex moutan nano-particles. Results Cortex moutan powder after super-micro-particle pulverization appears sphere or like-sphere, and its average size is 200nm~300nm. After the superfine grinding Paeonol dissolution increases 76.19% in comparison with without nano pulverization. The nanoparticle rest angle is θ=33°.The precipitation ratio of Cortex moutan powder with general grinding is 0.28 at 24h, and the precipitation ratio of its nano-power has been to 0.98 at 60min. Conclusion Paeonol dissolution, stability and fluidity of Cortex moutan nanoparticles were improved greatly and this nanoparticles is beneficial to industrial production for traditional Chinese medicine.