Dextran 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 molecular weight distribution of dextran. Some quality problems and clinical side effects induced by the disproportion of molecular weight, had become the bottleneck of study and application of dextran.In this study, Solvent Out Crystallization 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 molecular weight 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 ultrafiltration membranes with different molecular weight cut-off, it 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 ultrafiltration and reverse osmosis, 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.It distributes in torrid and temperate zone.It is abundant in flavone,tannis and Carotene etc which are bioactive components,so it is worthy of developing. This paper mainly conducted the extraction,purification and deodorant effect of polyphenols from persimmon peel.The results were showed that:The experiment used the traditional solvent extraction.Based on single factor tests,thogonal test was done.The optimum extraction conditions were that the concentration of acet was 50%,extraction temperature is 55℃,the extraction time was 105min and the ratio of the persimmon peel 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 antibacterial 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 deodorization 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 polyphenol,the sample was supposed to contain the polyphenol.By using the High performance liquid chromatography (HPLC),we selected three kinds of phenolics standards to determine small molecular phenolics in camelliia chryscath.The result showed that sample B-2 might containe gallic acid,the caffeine and the table caffeine.

Rosemary（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 rosmarinic acid, which had very strong antioxidant ability. In this paper, the technics of extraction and purification of rosmarinic acid （RosA） from rosemary and the antioxidant ability of the extract were studied.Firstly, the extraction of rosmarinic acid 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%. It 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 it 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,it has some important physiological functions in living creatures.In this paper,fresh mulberry leaf was studied with modern isolation technology and advanced methods of analysis and testing to extract,isolate and purify the mulberry leaf PPO;spectrophotometric analysis method and scanning electron microscopy were used to determine its structure,enzymatic properties and inhibitory effects of different inhibitors on it;regulatory mechanism of the PPO activity and immobilization 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 immobilization of mulberry leaf PPO were researched,4%sodium alginate,0.2%glutaraldehyde,0.2mol/L CaCl₂ used to immobilize the PPO, with an immobilization 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. Water quality 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 Bailuzhou wetland 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 Bailuzhou wetland.

Rice bran 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 mechanical 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 mechanical 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 macroporous adsorption resin 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 macroporous adsorption resin.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 it 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%.

Optically active (S)-1-phenyl-1,2-ethanediol((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 Candida parapsilosis 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 (S)-1-phenyl-1,2-ethanediol 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 it 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 %.

The N-feruloylserotonin (FS) and N-(p-coumaroyl)serotonin (CS) in safflower (Carthamus tinctorius L.) seed meal exhibited anti-oxidation, anti-inflammatory and anti-tumor effect, can be used in food, medicine, cosmetics and other fields. They have broad development prospects. In this paper, enrichment process of N-feruloylserotronin (FS) and N-(p-coumaroyl)serotonin (CS) in safflower (Carthamus tinctorius L.) seed meal has been researched systematically.Through single factor experiment and orthogonal experiment, the optimal technology of extracting the total phenylpropanoid amides of 5-hydroxytryptamine (PAHA) from meal was determined as follow: 60% ethanolas extraction solvent, extracted at 65℃for 90min, solid-liquid ratio of 1:20, grinding granularity of 35 mesh. The yield of PAHA was 0.784%, and the purity was 7.87%; The yield of the crude extraction was 9.96%, and the purity of FS, CS, FSG and CSG in the crude extraction were 3.82, 2.58, 0.53 and 0.94% respectively. The shrinking core model on PAHA extraction of safflower seed meal by 60% ethanol was established, based on the Fick’s first law. According to the experimental data, the model was solved and validated. The extraction rate-determining step of the process was the diffusion of solute through the extracted residual layer. And The extraction time t was in the good linear relationship with . The results proved that the model could be used to analyze and predict the process of extracting PAHA from safflower seeds by ethanol.The optimal enzymolysis conditions of PAHA glucoside were detemined by using the single factor experiment and response surface analysis:The crude extract concentration of PAHA 3.3g/L, pH=5.9, enzymolysis with 3.00% (g/g crude extraction)β-D-glucosidase under 47℃f or 10.5 hr. The conversation rate of N-(p-coumaroyl)serotonin-mono-β-D- glucopyranoside (CSG) and (N-feruloylserotronin)-mono-β-D-glucopyranoside (FSG) were 94.6 and 90.0% respectively, and the FS and CS content in the conversation product were 3.42 and 4.30%.After enzymatic hydrolysising PAHA crude extraction byβ-D-glucosidase, enrichment processes of FS and CS with D818 weak alkaline macropore anion ion-exchanger resin were studied. The optimal adsorption and desorption experiments on D818 resin were conducted to establish parameters as: FS and CS concentration in sample solution 30-50 and 25-41mg/mL, respectively, pH=9.1(for adsorption), sample flow rate 1.5 mL/min; eluent 80% ethanol, pH=3.2, flow rate 1 mL/min (for desorption). After one run of adsorption and desorption, the contents of FS and CS were increased from 3.42%, 4.30% to 23.6% and 19.4%, and the recoveries were 86.3% and 85.6%, respectively.Then used XDA-1 macroporous resin column chromatography for purifying poduct desorpted by D818 resin, the FS and CS crude mixture was received, and the contents of FS and CS were increased to 32.2% and 25.9%, and the recoveries were 85.4% and 83.3%, respectively. The process of gel column chromatography was researched as follow: eluted by petroleum ether/acetone=6:4,the flow rate 0.074cm/min,sample quality 0.012g/g gel,finally the contents of FS and CS in the product were 49.3% and 40.2%, and the recoveries were 91.3% and 92.7%, respectively.The monomers of FS and CS were detected respectively with HPLC-ELSD, IR and EIS-/MS, suggesting the chemical structure of CS and FS are unchanging after extraction, enzymolysis and purifying in enrichment process.

Zwitteronic surfactant has many excellent properties such as the low toxicity and stimulating to the skin and the eyes, good biologic degradation, low Krafft point, in addition it is able to bear the hard water and the high concentration electrolyte, basically no social effects of pollution. However, the inorganic salt producing in the zwitteronic surfactant synthesis process increases its viscosity, makes its pH not stable, decreases its stability, and effects its application performances. The difficulty purity of this surfactant limits its research and application. The separation techniques used in purification of this surfactant will promote the development of zwitteronic surfactant. In this paper, traditional purification methods were attempted to purify theα-capric betaine(α-CB). The electrodialysis desalination ofα-CB and the surface chemical properties and application performances ofα-CB were investigated. The main points were as follows:Recrystallization, ion exchange resin and extraction were applied to purify theα-CB. It was found that the fatty acid and the inorganic salt could be removed effectively by the method of recrystallization, which could be obtainedα-CB with mass fraction of 98%. Products identify and purity test were confirmed by IR, H-NMR, Ms, elemental analysis, melt point and surface tension curve.The electrodialysis desalination ofα-CB was investigated. The effects of the operation voltage, flow rate and pH of the solution on the electrodialysis desalination ofα-CB were investigated and the optimum operating condition was chosen through comparing desalination rate, the recovery ofα-CB and electricity consumption. The results showed that the inorganic salt existing in zwitterionic surfactant solution could be removed effectively. It was found that under an optimum operating condition, pH value 7.50 (at the isoelectric point ofα-CB), operation voltage 10 V, and flow rate 20 L/h, the desalination efficiency was 99% and the recovery ofα-CB was 82.7%. The electrodialysis desalination ofα-CB with constant current was investigated which made the recovery ofα-CB significantly increased. It was found that current 0.5 A, flow rate 20 L/h, pH value 7.50, the desalination efficiency was more than 99% and the recovery ofα-CB was 89.4%.The surface chemical properties and application performances ofα-CB were investigated.α-CB had good surface activity and foaming performance, the critical micelle concentration, minimum surface tension and foam height ofα-CB were tested to be 9.3×10-3 mol/L, 38.2 mN/m and 175 mm, respectively. The lime soap dispersing ability and hard water-resistance ofα-CB were superior to SDS, which exhibited excellent performance. Moreover, the application performance of soap addingα-alkyl betaine was investigated. The results showed thatα-alkyl betaine had synergism on wetting and hard water-resistance.

Lipases are important enzymes universally applied in industries; it is significant to screen new lipase resources and to characterize novel lipases for specific application.A strain of lipase producing microbial was isolated from nature. It was morphologically idntified as Geotrichum .sp and termed as Geotrichum sp. SYBC WU-3.The effect of inducing agents and suefactants on the lipase production of Geotrichum sp. SYBC WU-3 was investigated. The results showed that olive oil, nuts composite oil, Tween-20, Tween-80, and TritoonX-100 could promote lipase production by Geotrichum sp. SYBC WU-3, when nut complex oil and TritoonX-100 or olive oil and TritoonX-100 was added to the medium, a new isozyme named as LIP-1 could be induced, the constitutive lipas of Geotrichum sp. SYBC WU-3 was termed as LIP-2.The optimum medium for the yield of lipase was that containing peptone 5 g/L, NaH2PO4 3g/L, nuts oil 250 mL/L, Polyoxyethylene polyglycol ether 25 mL/L. The strain was cultivated at the temperature of 30℃for 72 h. The enzyme activity under the optimal condition was 3.2 folds of that under the condition before optimization.Isozyme LIP-1 and LIP-2 was purified by DEAE-32 chromatography, Sephadex G-100 gel filtration and two-phase extraction. LIP-1 and LIP-2 was purified 15.6 and 22 folds respctively. Their recovery was 11.15 %and 17.9 % respectively.Both LIP-1 and LIP-2 was cold-adapted alkaline lipase. Their optimal pH in 50 mmol/L pH 7.0 Tris-HCl buffer are 9.5. Both of them showed high lipolytic activity from pH 3.0 to 9.0. The optimal catalytic temperature of LIP-1 and LIP-2 was 20℃and 15℃respectively. LIP-1 was labile at 50℃, LIP-2 was labile at 40℃. LIP-1 had stronger ability to hydrolyte long chain fatty acid esters and LIP-2 had high hydrolysis ability to hydrolyte short chain fatty acid esters.