2.Purification of G6PD from erythrocytes by affinity chromatography
Journal of Vietnamese Medicine 1998;230(11):8-12
The procedure using affinity chromatography for purification of G6PD from rabbit erythrocytes has been developed. This method consists of three following steps: ion exchange chromatography on DEAE - Sephadex A50, precipitation with ammonium sulfate and affinity chromatography on 2'5' ADP - Sepharose 4B. The first and the second step are directly derived from the procedure of NguyÔn H÷u ChÊn. By our method, the accumulative purification of G6PD from rabbit erythrocytes was 10243 fold. A specific activity of 69.66 IU/mg protein and a final yield of 40% were obtained.
Isolation & purification
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Erythrocytes
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Chromatography
3.Purification and preliminary studies for application of lectin from haemolymph clam (Meretrix meretrix Linne) in medicine.
Journal of Medical and Pharmaceutical Information 1998;(1):34-38
A lectin from the haemolymph clam was purified by molecular sieving on sephadex- G75 column. The products have a high purity. The purity of lectin was assessed by SDS polyacrylamide gel. Two doublets band of apparent molecular weights of respectively about 14,000-17,000 and 20,000-23,000 Dalton were revealed upon staining with Coomassie blue. Optimum-pH of the lectin from the clams is pH 6,5-7,0 and 8,5-9,0. The lectins of the clams had very different specificity for hydratecarbons and glucoproteins
isolation & purification
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Lectins
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medicine
4.Selection of potential lactic acid bacteria from fermented Sumbawa mare’s milk as starter cultures
Ilmiyatus Safitri Devi ; Yoga Dwi Jatmiko
Malaysian Journal of Microbiology 2021;17(1):11-19
Aims:
The aim of this study was to screen lactic acid bacteria (LAB) isolates from fermented Sumbawa mare‘s milk that
meet the requirements as starter cultures, and to evaluate the effect of the selected starter culture in improving the
organoleptic quality of mare‘s milk fermentation.
Methodology and results:
The LAB isolates (13 isolates) derived from naturally fermented Sumbawa mare‘s milk were
firstly screened for acidification activity. Afterwards, the selected isolates were evaluated for the starter culture criteria
such as technological properties (proteolytic test, lipolytic test, and exopolysaccharide production), food safety test
(hemolytic test and antibiotic sensitivity test), antimicrobial activity test. The selected culture (SC) together with yogurt
starter cultures (YC) and combination between the selected isolate and a mixture of both (MC) were used to ferment
fresh mare’s milk. Six LAB isolates (DB7, BC10, DC4, BC9, DC10, and BC7) were obtained from the acidification
screening. Isolate BC10 was the most potential isolate as starter culture due to its ability in terms of acidification and
proteolytic activity, lack of lipolytic activity, no indication of pathogenic potency, as well as able to inhibit the growth of
Escherichia coli ATCC 25922. However, this isolate was resistant to antibiotics kanamycin, trimethoprim, and cinoxacin.
The isolate BC10 presented 99.99% sequence similarity with respect to Lactobacillus plantarum.
Conclusion, significance and impact of study
The selected starter culture (isolate BC10) was able to improve the
organoleptic quality of fermented mare‘s milk especially aroma compared to the other starter cultures. Therefore, L.
plantarum BC10 is a potential isolate to be used as starter culture for mare’s milk fermentation.
Lactobacillales--isolation &
;
purification
5.Phytochemical progress made in investigations of Angelica sinensis (Oliv.) Diels.
Jian-Ping MA ; Zhi-Bing GUO ; Ling JIN ; Ying-Dong LI
Chinese Journal of Natural Medicines (English Ed.) 2015;13(4):241-249
The phytochemical progress on Angelica sinensis (Oliv.) Diels over the past decades is summarized. Since 1970s, 165 chemical constituents, including phthalides, phenylpropanoids, terpenoids and essential oils, aromatic compounds, alkaloids, alkynes, sterols, fatty acids, and polysaccharides have been isolated or detected from the various parts of the title plant.
Alkaloids
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isolation & purification
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Alkynes
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isolation & purification
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Angelica sinensis
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chemistry
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Benzofurans
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isolation & purification
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Fatty Acids
;
isolation & purification
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Oils, Volatile
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isolation & purification
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Phytochemicals
;
isolation & purification
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Phytosterols
;
isolation & purification
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Polysaccharides
;
isolation & purification
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Propanols
;
isolation & purification
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Terpenes
;
isolation & purification
6.A new technique of extracting effective components from Chinese herb and natural plant--microwave assisted extraction, MAE.
Ying ZHANG ; Zhuo-yu YU ; Xiao-qin WU
China Journal of Chinese Materia Medica 2004;29(2):104-108
The extraction and separation of effective components from Chinese herb and natural plant is one of the most important processes of natural medicine production. Recently, more and more advanced techniques have been used in the field of modern Chinese medicines, for example, microwave-assisted extraction (MAE). Developed on the base of traditional extracting technology using by organic solvents, MAE is of higher extraction rate and efficiency and better extract quality, as well as few investment, simple equipment, wide adaptability, high selectivity, good fidelity and no pollution. This paper mainly reviews the action principle and characteristic of MAE and its application in the extraction of natural products such as flavonoids, glycosides, polysaccharides, terpenoids and essence. Otherwise, the developing foreground of MAE is also prospected here.
Drugs, Chinese Herbal
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analysis
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isolation & purification
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Flavonoids
;
isolation & purification
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Glycosides
;
isolation & purification
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Microwaves
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Plants, Medicinal
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chemistry
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Polysaccharides
;
isolation & purification
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Technology, Pharmaceutical
;
methods
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Terpenes
;
isolation & purification
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Triterpenes
;
isolation & purification
7.Analysis of the chemical constituents of Hedyotis diffusa.
Journal of Southern Medical University 2008;28(1):127-128
OBJECTIVETo analyze the chemical constituents of Hedyotis diffusa.
METHODSColumn chromatographies were used to isolate and purify the chemical constituents of this plant, and their structures were identified by spectral analysis and physicochemical properties.
RESULTS AND CONCLUSIONSeven compounds were isolated and identified as p-coumaric acid (I), methyl-p-coumarate (II), 2-formyl-5- hydroxymethylfuran (III), quercetin (IV), kaempferol (V), beta-sitosterol(VI) and daucosterol(VII), respectively, among which the compounds II and III were isolated from Hedyotis diffusa for the first time.
Antineoplastic Agents, Phytogenic ; isolation & purification ; Cinnamates ; isolation & purification ; Coumaric Acids ; isolation & purification ; Furans ; isolation & purification ; Hedyotis ; chemistry ; Kaempferols ; isolation & purification ; Propionates ; Quercetin ; isolation & purification
8.Progress in purification of human serum albumin.
Chinese Journal of Biotechnology 2002;18(6):761-766
Human serum albumin(HSA) has been used clinically to treat a number of diseases with high dosage. Extremely pure puoduct is required in large-scale production. Plasma-derived HSA(pHSA) has long been produced by precipitation methods. Among them cold ethanol precipitation is dominant. However, chromatographic purification of HSA has been increasingly studied in the last few years. Application of chromatography, especially ionexchange, affinity, and size-exclusion, has opened a new area in the production of pHSA. A new challenge is the purification of recombinant HSA(rHSA). A successful approach involves STREAMLINE expanded bed adsorption to direct capture the target product from the fermentation broth. This novel process eliminates the need to separate the cells by centrifugation or membrane filtration. Ion exchange chromatography and hydrophobic chromatography play a central role in the purification scheme. Integration with other chromatographic techniques such as size-exclusion, metal chelate, and affinity gives improved purification results. Though innovative, the purification of rHSA still needs further improvement and optimization to increase product purity and process recovery.
Humans
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Recombinant Proteins
;
isolation & purification
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Serum Albumin
;
isolation & purification
9.Depyrogenation in key manufacturing processes of Reduning injection.
Miao LI ; Yuling XU ; Juan SONG ; Yongxiang WANG ; Yingzhi PAN ; Zhengzhong WANG ; Wei XIAO ; Tao LIU
China Journal of Chinese Materia Medica 2011;36(6):663-665
OBJECTIVETo investigate the effect of removing bacterial endotoxin in the key processes of Reduning injection.
METHODThe content of bacterial endotoxins was detected by kenitic-turbidimetry and the removal efficacy was studied before and after using 0.8% of activated carbon and ultrafiltration with molecular weight cut-off of 10 x 10(3).
RESULTThe adsorption rate of bacterial endotoxins was 78.7% by using activated carbon, while the removal efficacy of bacterial endotoxins was 99.6% with ultrafiltration membrane at cut-off molecular weight 10 x 10(3).
CONCLUSIONThe key technology can effectively guarantee the safety of Reduning injection.
Adsorption ; Endotoxins ; isolation & purification ; Injections ; Pyrogens ; isolation & purification ; Ultrafiltration
10.Application of spiral disk column in high-speed counter-current chromatography for peptide and protein separation.
Chinese Journal of Biotechnology 2009;25(4):618-625
In order to improve the stationary phase retention of polar solvent systems and aqueous two-phase systems (ATPSs), we designed a multiple spiral disk assembly for type-J high-speed counter-current chromatography (HSCCC). The stationary phase retention was studied under different elution modes by using two solvent systems that contained 1-butanol-acetic acid-water (4:1:5, V/V/V) and polyethylene glycol (PEG) 1000-K2HPO4-water (12.5:12.5:75, W/W/W). The best retention was obtained in L-I-T, U-O-H, L-I-H three modes by pumping lower mobile phase from inner terminal (I) to outer terminal (O), and upper mobile phase from outer terminal (O) to inner terminal (I) at a relatively high flow rate. Meanwhile, the relationship between retention percentage of the stationary phase (Sf) and various parameters such as flow-rate (F), rotation speed (w) and column temperature (T) was also studied. Sf increased with the increase of w and decreased with the increase of F. Regression analysis showed a linear relationship between Sf and F1/2/w. The influence of T on Sf was not obvious between 20 degrees C and 40 degrees C, lower temperature than 20 degrees C was not suitable for viscous ATPSs. Acceptable resolutions were achieved when it was applied for the separation of dipeptides including Leu-Tyr and Val-Tyr by using 1-butanol-acetic acid-water (4:1:5, V/V/V) solvent system. The proteins including cytochrome C and myoglobin, lysozyme and myoglobin, and fresh chicken egg-white proteins were well separated by 12.5% PEG1000-12.5% K2HPO4-75% water (pH 9.0) and 16% PEG 1000-12.5% K2HPO4-71.5% water (pH 8.0) system.
Countercurrent Distribution
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instrumentation
;
methods
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Peptides
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isolation & purification
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Proteins
;
isolation & purification