Amorphophallus konjac is a monoecious and perennial herbaceous plant of Araceae.There are about 170 varieties in the world,and at least 21 varieties found in our country.There are mainly two varieties including Amorphophallus Albus and Amorphophallus Rivieri,in planting.The main component in the Amorphophallus konjac is(KGM),which has the function of shedding toxin,losing weight,relaxing bowels,cleaning stomach,controlling disease,diabetic and carcinoma,lowering blood-lipid and cholesterol,keeping salt-balance,adjusting immune function, regulating lipid metabolism,supplementing calcium and so on.Amorphophallus konjac was used as raw material in our experiment.The determination method of KGM,optimized conditions for purification of ,and possibility for KGM to replace PAM were studied.The results were as follows:1.Wavelength of determing KGM by spectrophotometer was detected.The results showed that: the maximum absorption band was 497nm,under this wavelength,the Relative error of testing is 0.3%.and under wavelength of 550nm,the counterparts are 2.1%,respectively,and under wavelength of 550nm the relative standard deviatetion of veficity is 1.13%。2. is feasible to purify the using Ethanol,and the optimized conditions were as follows:elution time 90rain,pH of the solution 4.5,concentration of ethanol 60%.Under those conditions,the maximum contents of KGM are higher up to 98.51%.3.The properties between highly purified KGM and PAM were caomapred.The results showed that was almost same in the solubility in between KGM and PAM,except a little bit more dissolving in benzene for KGM,but no for PAM.The similar transparency existed between the two but slightly better for PAM.Meanwhile,the sound stability for both temperature and pH value was observed.
Post about "Purification"
S-adenosyl-L-methionine（SAM）is an important metabolic intermediate in organism which is involved in many biochemical reaction.There are vast market for SAM as nutrient and drug.Our main task is carries on screening to yeast,and itsconditions to do research.The separation and purification of them were preliminary studied.SAM can be obtained by three ways:chemosynthesis,enzymatic conversion technique,as well as microbial ,of which is the primary way of industrial production.In our experiment one was choosen from four strains to produce SAM high saccharomycetes Sake,its output is the 1.71g/L.By the space-mutation Yeast Sake,was picked out 500 yeasts,then through refilter we obtained one to produce the SAM high strain,No.552 of the yeast than the originally and its production output increase 11.16%.By the fermentation,fermentation conditions Sake552 strain of did conduct a preliminary exploration. were studied and identified the best initial pH is 5.0,while SAM accumulation phase is 6.0 in shake flask;best fermentation temperature is 28℃;Best medium are 20%distilled potatoes, glucose 7%,yeast extract 1%,Urea 1.4%,L-Met 1%,KH2PO4 0.4%,K2HPO40.2%,MgSO4·7H2O 0.5%, CaCl2·2H2O 0.2%.The culture conditions of the Yeast Sake 552 in fermentor 5L were optimized, including the prescription of cultural medium,pH,dissolved oxygen,agitation speed,aeration,feeding strategy of complementarity-cultural medium in different stages.The SAM content of the Yeast Sake 552 reached 3g/L after fermentation for 4 days under the optimized conditions.We also has researched the separation and purification process of SAM and has established a preliminary technics.The procedure followed with that the cell suspension of pH2 to be frozen for 24h then to disrupt and then purify with weak acid cation exchange resin,to crystal and deposit with methanol,to centrifugal deposition to freeze-dry it finally we got the crude SAM.
In this study, theand protein in zizyphus jujube mill was used as the object, In the purification process, the was study on extracting discolor dialysis and de-protein for further study, the dissoluble protein was extracted dialysis electrophoresis high Performance Liquid Chromatograph, in order to offer theoretical foundation for produce and relevant research. The result is as follows:1.The optimal conditions for hot water was 1:16 of the ratio of sample to water,240 mins of at 100℃and one time, The rate was 5.94%;The optimal conditions for enzyme was pH4.5,60 mins of extraction at 60℃and 0.03% concentration of enzyme, The rate was 7.82%;The optimal conditions for alkali extraction was Na2CO3,1:20 of the ratio of sample to water,180 mins of extraction at 80℃, The rate was 6.82%.2.Choosing the optimal precipitator of methanol ethanol acetone, methanol would be the optimal precipitator, acetone is noxious but the ethanol is the cheapest, so we choose the ethanol; Sorite extraction method extrats fattiness; The separation of smaller molecules by 3000 Dalton dialysis membrane.3. De-color by activated carbon and hydrogen peroxide(H2O2). activated carbon is the best no side-effect and adsorbs some , polysaccharide would be oxidated by high consistence hydrogen peroxide. Sevag method was the optimal de-protein process.To sum up, the polysaccharide and the dissoluble protein was extracted and pured, and determined the molecular weight of the dissoluble protein.
Studies on Purification of Clinic Dextran with Solvent out Crystallization Assisted Membrane Separation Technology
is a bacterial polysaccharide produced from sucrose and usually used for clinical treatment as one kind of polysaccharide drugs. A traditional separating method with ethanol was hardly to control strictly for the distribution of dextran. Some quality problems and clinical side effects induced by the disproportion of , had become the bottleneck of study and application of dextran.In this study, assisted Membrane Separation (SOCMS) is a new separation method proposed for thepurification of dextran.Its purpose is to separate dextran and control itsmolecular distribution, and prepare dextran with high purity at the sametime.Primary research subjects and conclusions were summarized as follow:1.Chromatogram conditions to measure the ofdextran were determined.2. By using uniform experiments design, several factors that influenced the solvent out crystallization were investigated.3. Through analyzing purification effects of membranes with different molecular weight cut-off, showed that membranes of 30KD and 5KD were helpful to the purification of dextran.4. The determination of membrane separation parameters was established by single factor experiment.5. By connection of and , purification process was optimized.6. Cleanig condition of membranes were determined.7. The purity of the purified sample samples was identified by the ultraviolet spectrum. The result showed that there were no protein, nucleic acid, pigment and mannose within them. Preliminary studies on the structure of purified samples by the infrared spectrum illuminated that purified samples were the same material with the dextran. Analysis on thermodynamic properties by DSC showed that purified samples were much more similar in composition.
Persimmion belonging to Ebenaceae DiospyrosL is Deciduous tree.distributes in torrid and temperate zone. is abundant in flavone,tannis and Carotene etc which are bioactive components,so it is worthy of developing. This paper mainly conducted the ,purification and deodorant effect of s from .The results were showed that:The experiment used the traditional solvent .Based on single factor tests,thogonal test was done.The optimum conditions were that the concentration of acet was 50%,extraction temperature is 55℃,the extraction time was 105min and the ratio of the to solvent was 1:16.Under the optimal conditions the extraction yield could reach 71.16%.Through the absorption of static and dynamic curve of static studying in the six kinds of macroporous resin.Resoult showed that HP-20 resin had excellent absorption and desorption properties.The dynamic absorption conditions were flow speed 2BV/h,column density 2.91mg/mL and pH4.8. Through the research on gradient elution,it showed that:40%ethanol eluted the purity which could reach 32.04%.Through the bacterial experiments,three samples had effects of restraining bacterium.The sample B had better effect than the other two samples.The elimination effect enhanced with the concentration of sample increasing.The minimum inhibition concentration(MIC)of sample B on Escherichia coli,Staphy lococus and Saccharomyces cerevisiae were listed as follows:250mg/mL,125mg/mL,250mg/mL.The minimum sterilization quality fractions(MBC)were:500mg/mL,250mg/mL,500mg/mL.Combining apparatus with sensory analysis Method.The results showed that:the extract of Polyphenol had effect on ammonia,trimethlamin,sulfureted hydrogen,formaldehyde and indole.The sample B had the best effect on trimethlamin and the worst effect on formaldehyde.This might relate Polyphenol purity,the structure and character of the smelt substance.Apparatus and sensory analysis methods could show the effect of exactly.Through the further purification by Sephdaex LH-20,the Polyphenol purity of sample B-2 was 53.64%.The deodorant rate of it on trimethlamine was 62.37%(2mg/mL).Samples were detected by UV spectrum.After compared to the characterristic absorption of ,the sample was supposed to contain the polyphenol.By using the High performance liquid chromatography (HPLC),we selected three kinds of phenolics standards to small molecular phenolics in camelliia chryscath.The result showed that sample B-2 might containe gallic acid,the caffeine and the table caffeine.
The Study on Extraction of Rosmarinic Acid from Rosemary and Their Antioxidant Ability of the Rosemary Antioxidant
（Rosmarinus officinalis L.）, an evergreen shrub, which is a natural spiceberry of the family Labiatae, contained many kinds of bio-active substances. One of these substances was , which had very strong antioxidant ability. In this paper, the technics of and purification of （RosA） from rosemary and the antioxidant ability of the extract were studied.Firstly, the of from rosemary with microwave and ultrasonic wave was studied. The results showed that the optimal extracting technological conditions were as follows: 20 mesh（comminuted size, mesh）, 15% （ethanol concentration, v/v）, ratio of liquid to material （5:1）, 10 min（the time of ultrasonic cells broken）, 540w （the power of microwave）,4 times （extracting times） and 6 min each time. Under these conditions, the extracting rate of rosmarinic acid could reach 94.54%. showed that this method had the advantages of short extracting time, high yield and low-cost for extraction of rosmarinic acid.Secondly, the optimal conditions of purification were studied experimentally. The extraction liquid was clarified by ceramic micro-filter and used X-5 macroporous absorption resin to concentrate rosmarinic acid. The optimal adsorption conditions were determined: When the adsorption speed was 4 mL·min-1, the resin could absorb 1500 mL extraction liquid , and 1300mL 75% ethanol was used to desorb.Thirdly, the gained concentration solution was purified by normal butanol, and the best condition of extraction was: When the solid concentration was 14.29%, pH=3, extraction time was 20min, the volume ratio of solution to normal butanol was 1:1.5, the extraction times was 2.Finally, the crystallization method was used to the further purify to gain the pure products. The qualitative analysis by HPLC showed that the content of the final rosmarinic acid products was 96.82%（w/w）.Furthermore, the antioxidant activities of rosemary extract was studied. The rosmarinic acid products had a good antioxidant ability though showed slightly lower reducing power and scavenging ability on superoxide radical(·O-2) than ascorbic acid. And in order to make the best use of rosemary, the lipid-soluble antioxidant was extracted from the material dregs.All in all, the purity of rosmarinic acid products was high. The whole technics were short and easy to undertake in factory which had a good future in industrial application.
Polyphenol oxidase（PPO）is widely distributed in microorganism,plant, animal and human body,has some important physiological functions in living creatures.In this paper,fresh was studied with modern isolation technology and advanced methods of analysis and testing to extract,isolate and purify the PPO;spectrophotometric analysis method and scanning electron microscopy were used to determine its structure,enzymatic properties and inhibitory effects of different inhibitors on ;regulatory mechanism of the PPO activity and were also discussed,in order to empolder the mulberry leaf PPO and increases its economic value.Main results were:The optimal extraction technics for mulberry leaf PPO were ascertained, 1.0%Tween-80,pH 6.0,extraction time were 2h,the ratio of solid to solvent were 1:2.The purification processes for mulberry leaf PPO were discussed,the classification deposition was disposed using 30%and 80%saturation of ammonium sulfate to isolate the impurity protein from PPO and increase the PPO yield.The Sephadex G-75（16×800mm）column,velocity of flow in elution was 15mL/h to further purify the crude PPO.First elution peak value was mulberry leaf PPO.The mulberry leaf PPO structure,molecular weight and enzymatic properties were determined,the enzyme was fibrous protein observed by scanning electron microscopy;its molecular weigh was 47Kd;the optimum substrate was pyrogallic acid,its Km value was 0.093mol/L;and its optimum pH and temperature were 7.0 and 40℃respectively;with the same concentrations of different inhibitors,the total inhibitory effects were:ascorbic acid＞sodium hydrogen sulfite＞citric acid＞magnesium chloride＞sodium chloride.The of mulberry leaf PPO were researched,4%sodium alginate,0.2%glutaraldehyde,0.2mol/L CaCl2 used to immobilize the PPO, with an time of 2h and crosslinking time of 4h,could make the immobilized PPO activity reach its maximum.After its immobilization,the optimum pH was 6.0,and the enzyme showed a good stability between the pH from 5.0 to 7.0;the optimum temperature was 50℃for the enzyme,and the enzyme became less susceptible to the temperature,the temperature did not more affect to the enzymatic activity.
Aquatic plants are critical components of aquatic ecosystem. Their successful structures made up of lake aquatic plant community, and took a great role of water purification. Two parts were dealt with in the paper. The first was the investigation to the functions of city lake aquatic plants on water purification. And the second was making a reconstruction plan for a wetland aquatic ecosystem based on a typical city wetland (Nanjing Bailuzhou Park wetland). An contrast analyses of the water qualities pre and post the aquatic plant restoration was made from experiments. The research results could make a large contribution to the conservation projects of lake and wetland in city and provide both theoretical and technical supports for water purification.In order to compare the ability of aquatic plants on water quality improvement in cities, using Xuanwu Lake, Mochou Lake and Bailuzhou Park in Nanjing as research objecte, the water purification ability resulted from effective structures of aquatic plants community were discussed in the paper according to the study of water indexes, lake silt and aquatic plant compositions. The results showed:Dissolved Oxygen (DO), Biological Oxygen Demand (BOD), Chemical Oxygen Demand (COD), Total Nitrogen (TN), Total Phosphorus (TP) etc. were the main factors to reflect water pollution in urban lakes. The water quality change in a year was in the order of summer> spring> autumn> winter.At the same time and same lake but different sampling points, there were significant differences between water qualities.was usually much better where there was plant growth and a little artificial disturbance. Comparing and analyzing the purifications of communities of submerged macrophyte, floating plants and emerged plants, the results showed that submerged macrophyte had better absorption ability to TN and TP and could effectively improve water environment. There was in the order of submerged macrophyte > floating plant > emerged plant.For the collocation of communities of aquatic plants and sight rebuilding in the urban classic lake, setting as an example, the reasonable kinds of water plants with applications of the pre-research was choosed, and a project of the collocation of plant communities and sights rebuilding was applied by combining zoology functions of .
Study on Separation and Purification of γ-Aminobutyric Acid Accumulated Through Rice Bran Glutamate Decarboxylase
is a by-product of rice processing,with high annual yield but very low development and utilization level.The enzyme activity of Glutamate decarboxylase（GAD） in rice bran is one of the highest among all kinds of plants.Based on the enzymatic properties of GAD in rice bran,this paper studied the accumulation ofγ-aminobutyric acid（GABA） through rice bran GAD.In order to activate rice bran GAD,two process conditions were designed to simulate different plant adverse conditions:the direct method simulated adverse conditions such as thermal stimulation,hypoxia and salt stress;the enzyme extracted method simulated adverse conditions such as thermal stimulation,hypoxia,salt stress and stimulation.The separation and purification conditions of GABA from GABA accumulated solution were also studied.The specific contents include:process conditions comparison between direct method and enzyme extracted method,immobilized rice bran GAD to produce GABA,separation and purification of GABA from GABA accumulated solution.The two process conditions of direct method and enzyme extracted method were compared and optimized.The maximum yield of direct method was 15.37mg·（g rice bran）-1 after doing orthogonal test,while the maximum yield of enzyme extracted method was 29.89mg·（g rice bran）-1.Among the above adverse conditions,the stimulation had the strongest activated effect on rice bran GAD.According to the immobilized enzyme technology,rice bran was immobilized with healthy and non-toxic sodium alginate to produce GABA.The optimal immobilized conditions were:6.0%rice bran,2.5%sodium alginate,1h hardening time and 1.0%CaCl2. The study on enzymatic properties of immobilized rice bran GAD showed that the optimum temperature moved to 45℃from 40℃.The enzymatic activity recovery of immobilized rice bran GAD was 51.00%.The GABA content of GABA accumulated solution prepared by immobilized rice bran GAD was higher than the unimmobilized rice bran GAD.Ethanol precipitation was used to separate GABA from the accumulated solution.When the ethanol concentration was 75%,more than 90%of protein and carbohydrate was precipitated while the lose rate of GABA was low.So this method can be used to preliminarily separate GABA accumulated solution.LSA-8B is an efficient resin with high selectivity,which showed good adsorption of protein,pigment and carbohydrate and the recovery rate of GABA is high.So the LSA-8B resin can be used to GABA accumulated solution purification.The adsorption process is a heat absorption one, mainly expressed as a physical adsorption process.The adsorption process can speed up as the temperature raise.The influencing factors of adsorption process were studied,and the optimum conditions were as follows:using the original sample concentration,adjusted pH value to 5.0,and feeding rate 2BV·h-1,adsorbing protein and pigment;then adjusted pH value to 8.0,feeding rate 1BV·h-1,adsorbing carbohydrate and else.Ethanol in alkaline condition was an efficient regenerant to LSA-8B .Dealing with purification,the GABA content of GABA accumulated solution prepared by enzyme extracted method increased from 4.76%to 13.90%,the recovery rate of GABA was 75.52%.The GABA accumulated solution prepared by immobilized rice bran GAD contains very little protein and carbohydrate,so was purified by LSA-8B resin directly without ethanol precipitation operation.The GABA content increased to 4.04%from primary 3.06%,and the recovery rate of GABA was 98.06%.
Improvement of the Whole Cell Catalytic Efficiency of Asymmetric Production of (S)-1-phenyl-1,2-ethanediol by in Situ Resin Adsorption
Optically active, ((S)-PED) has special bioactivity and photoelectromagnetism performance, which is a important chiral intermediate in the synthesis of pharmaceuticals, agrochemicals, and fine chemicals. Preparation of optically pure (S)-PED by biocatalyst has an advantage of mild reaction conditions, high stereoselectivity,high regioselectivity and high chemical selectivity. In this paper, the asymmetric synthesis of (S)-PED catalyzed by CCTCCM203011 in aqueous phase was investigated. For the aim of laying a foundation to prepare (S)-PED on industrial scale, the initial reaction substrate concentration, reusing of biocatalyst, extracting and purification of (S)-PED were studied.In situ adsorption technology was used to increase the initial reaction substrate concentration. The results showed that by adding NKAⅡadsorbent resin in reaction system, the initial substrate concentration was increased. A formula was established about the resin addition amount along with the changing of substrate concentration, according to the formula, the PED concentration in aqueous phase could be controlled in best level. Thus, the high substrate concentration and high reaction speed could be achieved. Then 0.74 g NKAⅡadsorbent resin was added to system under an increased substrate concentration of 40 g/L with a preadsorption for 2 h before reaction. After reacting for 108 h, the optical purity and the yield of (S)-enantiomer reach 98.1 %e.e. and 88.5 % respectively. Thus, the (R,S)-PED concentration increased about 166.7 % than ever before.The optimization of temperature, pH, and agitation to reaction system containing resin were studied. The optimum reaction conditions were obtained as 30℃, pH 8.0, shaking speed 120 r/min. Under such conditions, the reaction time was shortened, and the product also had good level when initial (R,S)-PED concentration increased from 40 g/L to 50 g/L. At the same time, the effect of addition of xylose and yeast extract on stability of whole cell was also studied. Xylose and yeast extract could increase catalyzing batch of whole cell. When reaction conditions and addition amount of xylose and yeast extract were optimized, the optical purity and yield of (S)-PED could reach 97.8 % and 80.6 % respectively after reacting for 48 h in second reaction batch, and e.e. reach 95.0 % in the third batch, under the substrate concentration of 30 g/L. Metabolic activity was increased obviously after addition of xylose and yeast extract.The extraction and purification of , was studied. From 6 kinds of macroporous adsorbent resins, NKAⅡresin was selected to adsorb (S)-PED. The saturated adsorption capacity of NKAⅡresin was 250 mg/(g wet resin) while the time for to achieve saturation adsorption was 2 h. Besides, the adsorption isotherm data of NKAⅡresin could well fit the Freundlich equation. Ether was selected as a suitable solvent to recrystallization. 1 L solution containing (S)-PED was adsorbed by the resin column with a flow rate of 1.5 BV/h, desorbed by 2 BV ethyl acetate, decolored by 3 %(mass concentration)active carbon by 1 h, recrystallized with about 100 mL ether. When (S)-PED was extracted and purified under this condition, the total yield of (S)-PED was 68.7 %, with mass fraction 98.9 % from 1.4 %, and optical purity 99.0 %.