1.Scale-up preparation of phycoerythrin from Porphyra haitanensis.
Chunxia LI ; Daiyuan YAN ; Jing NI ; Ziye GUO ; Chun'er CAI ; Peimin HE
Chinese Journal of Biotechnology 2011;27(4):614-619
We developed large-scale preparation of phycoerythrin from Porphyra haitanensis, a main economic red algae in China. Firstly, P. haitanensis thallus was broken by using "swelling and smash" method. Then times of grads ammonium sulfate precipitation applied to the crude extraction were compared. Desalted solution was further purified with one-step chromatography using hydroxyapatite and properties on spectrum and molecular weight were identified finally. The results indicated that after four times of ammonium sulfate precipitation (15%, 50%, 10% and 40%), the absorption spectrum purity of P. haitanensis achieved 0.9 (A564/A280), and 507.82 mg phycoerythrin (A564/A280 > 3.2) was obtained from 7 kg fresh algae after further hydroxyapatite chromatography. This research provides a potential way for preparation of phycoerythrin in large sclae.
Ammonium Sulfate
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chemistry
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Chromatography
;
methods
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Phycoerythrin
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isolation & purification
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Porphyra
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chemistry
2.Elevation of alul-induced frequencies of chromosomal aberrations and sister chromatid exchanges in CHO cells substituted with 5-bromodeoxyuridine by ammonium sulfate.
In Jang CHOI ; Ihn Hwan LEE ; Sung Ik CHANG
Korean Journal of Anatomy 1991;24(1):61-69
No abstract available.
Ammonium Compounds*
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Ammonium Sulfate*
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Animals
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Bromodeoxyuridine*
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CHO Cells*
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Chromosome Aberrations*
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Cricetinae
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Humans
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Siblings*
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Sister Chromatid Exchange*
3.Partial Purification of the L-Asparaginase from Escherichia Coli and a Study on its Enzymatic Properties.
Korean Journal of Urology 1973;14(3):155-163
1. Based on the differences in solubility in ammonium sulfate and activity as a function of pH, two L-asparaginases, EC-1 and 2, were partially purified from Escherichia coli 0112. 2. Both L-asparaginases, EC-1 and 2, were highly activated with low concentration of their substrate, L-aspartgine, but inactivated with high concentration of it. 3. Activities of L-asparaginases, EC-1 and 2, were inhibited with the product, L-aspartic acid in proportion to its concentration 4. Both of them were denatured by urea, EC-1 being denatured completely with 6M and EC-2 with 8M urea, showing their spatial conformational difference, since the latter proved to be a little more resistant to urea than the former.
Ammonium Sulfate
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Aspartic Acid
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Escherichia coli*
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Escherichia*
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Hydrogen-Ion Concentration
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Solubility
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Urea
4.Cross-validation of the osmotic pressure based on Pitzer model with air humidity osmometry at high concentration of ammonium sulfate solutions.
Xiao-Lan WANG ; Ting-Ting ZHAN ; Xian-Cheng ZHAN ; Xiao-Ying TAN ; Xiao-You QU ; Xin-Yue WANG ; Cheng-Rong LI
Acta Pharmaceutica Sinica 2014;49(1):101-105
The osmotic pressure of ammonium sulfate solutions has been measured by the well-established freezing point osmometry in dilute solutions and we recently reported air humidity osmometry in a much wider range of concentration. Air humidity osmometry cross-validated the theoretical calculations of osmotic pressure based on the Pitzer model at high concentrations by two one-sided test (TOST) of equivalence with multiple testing corrections, where no other experimental method could serve as a reference for comparison. Although more strict equivalence criteria were established between the measurements of freezing point osmometry and the calculations based on the Pitzer model at low concentration, air humidity osmometry is the only currently available osmometry applicable to high concentration, serves as an economic addition to standard osmometry.
Ammonium Sulfate
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chemistry
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Freezing
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Humidity
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Osmolar Concentration
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Osmometry
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methods
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Osmotic Pressure
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Solutions
5.Stability of the hydrogenase from Tetraselmis subcordiformis and its preliminary purification.
Fei YAN ; Zhao'an CHEN ; Xupeng CAO ; Hongbin LU ; Song XUE ; Wei ZHANG
Chinese Journal of Biotechnology 2010;26(7):1003-1008
Tetraselmis subcordiformis, a marine green alga, can produce hydrogen by photobiologically hydrolyzing seawater with hydrogenase. In this study, the preliminary purification of the enzyme was explored by ammonium sulfate precipitation, and the impact of sodium dithionite, beta-mercaptoethanol and glycerol on the enzyme stability during the process was investigated. The experimental results illustrated that sodium dithionite provided significant protection on the hydrogenase by depleting oxygen, while glycerol, a protectant against the structure instability of the enzyme, also presented protection. Crude enzyme with specific activity of 0.557 U/mg protein was extracted using 60%-70% saturated ammonium sulfate solution supplemented with 200 mmol/L sodium dithionite and 5% glycerol, and the hydrogenase recovery yield was about 30%.
Ammonium Sulfate
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chemistry
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Chemical Precipitation
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Chlorophyta
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enzymology
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Enzyme Stability
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Hydrogen
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metabolism
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Hydrogenase
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isolation & purification
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metabolism
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Seawater
6.Isolation and purification of alpha-glycerophosphate oxidase in a polyethylene glycol/(NH4 )2SO4 aqueous two-phase system.
Yao MENG ; Jiagui JIN ; Shuangfeng LIU ; Min YANG ; Qinglian ZHANG ; Li WAN ; Kun TANG
Journal of Biomedical Engineering 2014;31(1):136-141
Alpha-glycerophosphate oxidase (alpha-GPO) from Enterococcus casseli flavus was successfully isolated and purified by using polyethylene glycol (PEG)/(NH4)2SO4 aqueous two-phase system (ATPS). The results showed that the chosen PEG/(NH4)2SO4 ATPS could be affected by PEG molecular weight, pH, concentration of PEG and (NH4)2SO4, and inorganic salt as well as additional amount of crude enzyme. After evaluating these influencing factors, the final optimum purification strategy was formed by 16.5% (m/m) PEG2000, 13.2% (m/m) (NH4)2SO4, pH 7.5 and 30% (m/m) additive crude enzyme, respectively. The NaCl was a negative influencing factor which would lead to lower purification fold and activity recovery. These conditions eventually resulted in the activity recovery of 89% (m/m), distribution coefficient of 1.2 and purification fold of 7.0.
Ammonium Sulfate
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chemistry
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Glycerolphosphate Dehydrogenase
;
chemistry
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isolation & purification
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Molecular Weight
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Polyethylene Glycols
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chemistry
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Water
8.Use biologic fibrin adhesive in otologic surgery: compared with ammonium sulfate fibrin adhesive and tissell®.
Hyung Chul LEE ; Mi Gyeung YANG ; Mun Heum PARK
Yeungnam University Journal of Medicine 1991;8(1):127-135
Successful middle ear surgery requires the availability of al safe, effective bonding material. Side effect caused by synthetic materials have led to the use of biologic adhesive, However, they carry the risk of transmission of infectious diseases if they are prepared from pooled human blood. The adhesive strength of ammonium sulfate fibrin adhesive produce an adhesive strength that is half that of the homologous commercial product. It is, however, good enough for use in several otolaryngological operations, tympanoplasty, facial nerve repair, reconstruction of ossicles. Reconstruction of posterior wall of ear canal and obliteration of frontal sinus and mastoid antrum using bone dust.
Adhesives
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Ammonium Compounds*
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Ammonium Sulfate*
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Communicable Diseases
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Dust
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Ear Canal
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Ear, Middle
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Facial Nerve
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Fibrin Tissue Adhesive*
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Fibrin*
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Frontal Sinus
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Humans
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Mastoid
;
Tympanoplasty
9.Chemical Burn due to a Hair Bleaching Agent.
Bo Suk KIM ; Se Young PARK ; Kun PARK ; Seok Don PARK
Korean Journal of Dermatology 2010;48(9):814-816
Several hair dressing procedures, for example, bleaching and dyeing of the hair, are currently being frequently performed for cosmetic purposes. While allergic contact dermatitis due to hair dressing products has often been described, only a few cases of burns caused by hair dressing chemicals have been reported. A 6-year-old girl presented with an 8x6 cm sized round ulcerative lesion on the occipital area after hair bleaching with ammonium persulfate mixed with hydrogen peroxide. The skin biopsy specimen showed epidermal necrosis and sclerosing change in the dermis. She was referred to another burn center and then she was treated with skin grafting by a plastic surgeon. To the best of our knowledge, this is the first reported case of chemical burn due to a hair bleaching agent in the Korean dermatologic literature.
Ammonium Sulfate
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Bandages
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Biopsy
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Burn Units
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Burns
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Burns, Chemical
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Child
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Cosmetics
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Dermatitis, Allergic Contact
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Dermis
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Hair
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Humans
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Hydrogen Peroxide
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Necrosis
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Quaternary Ammonium Compounds
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Skin
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Skin Transplantation
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Ulcer
10.Purification of Native Ag85 Complex, 38-kDa and MTB12 Protein Antigens from the Culture Filtrate of Mycobacterium tuberculosis.
Ji Sook LEE ; Tae Hyun PAIK ; Yung Choon YOO ; Junglim LEE ; Arum SHIN ; Chang Hwa SONG ; Eun Kyung JO ; Hwa Jung KIM ; Jung Kyu PARK
Journal of Bacteriology and Virology 2006;36(4):211-220
The purification of immunodominant native protein antigens from the culture filtrates of Mycobacterium tuberculosis is needed for the development of new vaccines and immunodiagnostic reagents against tuberculosis. In the present study, we conducted large scale purification of well-known secreted antigens, Ag85 complex, 38-kDa, and MTB12, from the culture filtrate proteins (CFPs) prepared from M. tuberculosis H37Rv grown as a surface pellicle on synthetic Sauton medium. The protein and antigen concentrations of culture filtrates were sufficiently increased after 6 week of culture. The MTB12 antigen was detected as early as 1 week of culture, and Ag85 complex and 38-kDa antigen were detected after 2 and 3 week of culture, respectively, by immunodiffusion with specific antiserum against 100-fold concentrated culture filtrates. For large-scale purification, the six-week-culture filtrates of M. tuberculosis H37Rv diluted 2.5-fold with 20 mM Tris-HCl, (P)H 8.3 were subjected to anion-exchange chromatography. The CFPs were eluted with 100 mM NaCl-20 mM Tris-HCl, pH 8.3 and concentrated by ultrafiltration. The concentrated CFPs were fractionated with ammonium sulfate, and followed by hydrophobic interaction chromatography and anion-exchange chromatography (FPLC). Eventually, 10 mg of Ag85 complex, 0.56 mg of 38-kDa, and 1.81 mg of MTB12 antigens were purified from 1 liter of the six-week-culture filtrates of M. tuberculosis H37Rv which contained 307.81 mg of protein of culture filtrate.
Ammonium Sulfate
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Chromatography
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Hydrogen-Ion Concentration
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Hydrophobic and Hydrophilic Interactions
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Immunodiffusion
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Indicators and Reagents
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Mycobacterium tuberculosis*
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Mycobacterium*
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Tuberculosis
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Ultrafiltration
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Vaccines