• 제목/요약/키워드: Production and Purification

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Studies on the Production of Thermostable Amylase. Part 1. Optimal Culture Conditions and Purification of Enzyme. (내열성 Amylase의 생산에 관한 연구 (제1보) 최적배양조건과 효소의 정제)

  • 오두환;이강표;변유량;유주현
    • Microbiology and Biotechnology Letters
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    • v.9 no.2
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    • pp.91-97
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    • 1981
  • A thermophilic soil isolate Bacillus sp. Y-127 was selected for the production of thermostable amylase. The strain was used for the enzyme production and the thermostable amylase was characterized. The optimum cultural conditions for the enzyme production were 6$0^{\circ}C$ at pH 7.0 for 32 hours using a mineral medium containing 2% soluble starch and 0.2% yeast extract. The extra-cellular enzyme was purified about 123-folds with about 6% recovery. The purified enzyme was stable at pH between 4.0 and 7.0, and temperature up to 6$0^{\circ}C$.

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A Convenient Manufacturing Method for Mass Production of EGCG Rich Green Tea Extract (Epigallocatechin Gallate 고함유 녹차추출물의 제조공정 개선)

  • Seo, Eun Hye;Kim, Eun Jeong;Cheon, Seong Bong;Yoon, Min Ji;Choi, Sang Un;Ryu, Geon-Seek;Ryu, Shi Yong
    • Korean Journal of Pharmacognosy
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    • v.50 no.3
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    • pp.198-204
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    • 2019
  • A facile and convenient method was developed for the mass production of epigallocatechin gallate (EGCG) rich green tea extract (Er-GTE). The Er-GTE was successfully obtained from the crude water extract of green tea by the combination of two step purification, i.e., a simple adsorption process on the cation exchange resins (Trilite SCR-B) followed by the chromatography with Diaion HP-20 resins. The green tea extract produced by water extraction under $45^{\circ}C$ was subjected to adsorb on the strongly acidic cation exchange resin, Trilite SCR-B. The eluate passed through the resin was reabsorbed on Diaion HP-20 resin, which was subjected to elute with a mixture of water and alcohol by conventional chromatographical manner. The EGCG content in Er-GTE was estimated above 97% by HP-LC analysis and the newly developed method was regarded as the most suitable and appropriate process for the mass production of epigallocatechin gallate rich green tea extract (Er-GTE).

Separation and Purification of Angiotensin Converting Enzyme Inhibitory Peptides Derived from Goat's Milk Casein Hydrolysates

  • Lee, K.J.;Kim, S.B.;Ryu, J.S.;Shin, H.S.;Lim, J.W.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.5
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    • pp.741-746
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    • 2005
  • To investigate the basic information and the possibility of ACE-inhibitory peptides for antihypertension materials, goat's caisin (CN) was hydrolyzed by various proteolytic enzymes and ACE-inhibitory peptides were separated and purified. ACE-inhibition ratios of enzymatic hydrolysates of goat's CN and various characteristics of ACE-inhibitory peptides were determined. ACE-inhibition ratios of goat's CN hydrolysates were shown the highest with 87.84% by pepsin for 48 h. By Sephadex G-25 gel chromatograms, Fraction 3 from goat's CN hydrolysates by pepsin for 48 h was confirmed the highest ACE-inhibition activity. Fraction 3 g and Fraction 3 gh from peptic hydrolysates by RP-HPLC to first and second purification were the highest in ACE-inhibition activity, respectively. The most abundant amino acid was leucine (18.83%) in Fraction 3 gh of ACE-inhibitory peptides after second purification. Amino acid sequence analysis of Fraction 3 gh of ACE-inhibitory peptides was shown that the Ala-Tyr-Phe-Tyr, Pro-Tyr-Tyr and Tyr-Leu. IC$_{50}$ calibrated in peptic hydrolysates at 48 h, Fraction 3, Fraction 3 g and Fraction 3 gh from goat's CN hydrolysates by pepsin for 48 h were 29.89, 3.07, 1.85 and 0.87 g/ml, respectively. Based on the results of this experiment, goat's CN hydrolysates by pepsin were shown to have ACE-inhibitory activity.

Purification and Production Conditions of Antimicrobial Compound from Methylotrophic Actinomycetes MO-16 (Methanol 자화방선균 MO-16으로부터 항균성 물질의 정제 및 생산조건)

  • 김현수;이정수
    • Microbiology and Biotechnology Letters
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    • v.27 no.5
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    • pp.391-398
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    • 1999
  • A methylotrophic actinomycetes strain MO-16, which produce the antimicrobial compound, was isolated from soil and supposed as Amycolatopsis sp. based on taxonomic studies. The cell-free extract of methanol-grown strain MO-16 showed dehydrogenase activity for methanol and formaldehyde when various electron acceptors were added for oxidation. On the other hand, methanol did not affect the production of antimicrobial compounds, and organic nitrogen sources such as corn steep liquor and peptone were better than inorganic nitrogen sources. These compounds showed broad antimicrobial spectrum to the tested strains such as bacteria and yeast. The antimicrobial comounds were very stable under heat(121$^{\circ}C$), acid(pH2.0), alkali(pH11.0) treatments. These compounds were isolated by ethylacetate extract, silica gel column chromatography and reverse phase HPLC. Two compounds(peak 1 and 2) were detected as antimicrobial compounds through the HPLC analysis. The peak 2 was purified as a single compound and revealed a 98% purity.

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Production, Purification, and Characterization of Taxol and 10-DABIII from a new Endophytic Fungus Gliocladium sp. Isolated from the Indian Yew Tree, Taxus baccata

  • Sreekanth, D.;Syed, A.;Sarkar, S.;Sarkar, D.;Santhakumari, B.;Ahmad, A.;Khan, M.I.
    • Journal of Microbiology and Biotechnology
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    • v.19 no.11
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    • pp.1342-1347
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    • 2009
  • We have isolated endophytic fungi from the Indian yew tree, Taxus baccata, and then screened for taxol production. Out of the 40 fungal cultures screened, one fungus Gliocladium sp. was found to produce taxol and 10-DABIII (10-deacetyl baccatin III). These compounds were purified by TLC and HPLC and characterized using UV-spectroscopy, ESI-MS, MS/MS, and proton NMR. One liter of Gliocladium sp. culture yielded $10\;{\mu}g$ of taxol and $65\;{\mu}g$ of 10-DABIII. The purified taxol from the fungus showed cytotoxicity towards cancer lines HL-60 (leukemia), A431 (epidermal carcinoma), and MCF-7 (breast cancer).

Recovery Processes of Organic Acids from Fermentation Broths in the Biomass-Based Industry

  • Li, Qian-Zhu;Jiang, Xing-Lin;Feng, Xin-Jun;Wang, Ji-Ming;Sun, Chao;Zhang, Hai-Bo;Xian, Mo;Liu, Hui-Zhou
    • Journal of Microbiology and Biotechnology
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    • v.26 no.1
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    • pp.1-8
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    • 2016
  • The new movement towards green chemistry and renewable feedstocks makes microbial production of chemicals more competitive. Among the numerous chemicals, organic acids are more attractive targets for process development efforts in the renewable-based biorefinery industry. However, most of the production costs in microbial processes are higher than that in chemical processes, among which over 60% are generated by separation processes. Therefore, the research of separation and purification processes is important for a promising biorefinery industry. This review highlights the progress of recovery processes in the separation and purification of organic acids, including their advantages and disadvantages, current situation, and future prospects in terms of recovery yields and industrial application.

Production of Recombinant Proteins as Immuno-Analytical Markers of Genetically-Modified Organisms (GMO)

  • Hwang, Ok-Hwa;Park, Hyuk-Gu;Paek, Eui-Hwan;Paek, Se-Hwan;Park, Won-Mok
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.783-788
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    • 2004
  • Marker proteins of genetically-modified organisms (GMO) and their antibodies were prepared and characterized as major components of an analytical system. We selected two GMO markers, neomycin phosphotransferase II and 5- enolpyruvylshikimate-3-phosphate synthase, and produced them from E. coli employing genetic recombination technology. After purification, their structural conformation and binding affinities to the respective antibodies were characterized. The results showed that the recombinant proteins were identical with commercially obtained reference proteins. We further used them as immunogens to raise polyclonal antibodies capable of discriminating GMO containing protein from non-GMO. Well-characterized marker proteins and antibodies will be valuable as immunoreagents in constructing analytical systems such as biosensors and biochips to measure quantities of GMO.

Medium Optimization and Application of Affinity Column Chromatography for Trypsin Production from Recombinant Streptomyces griseus

  • Chi, Won-Jae;Song, Ju-Hyun;Oh, Eun-A.;Park, Seong-Whan;Chang, Yong-Keun;Kim, Eung-Soo;Hong, Soon-Kwang
    • Journal of Microbiology and Biotechnology
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    • v.19 no.10
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    • pp.1191-1196
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    • 2009
  • The production of Streptomyces griseus trypsin (SGT) by S. griseus IFO13350 transformed with the expression vector pWHM3-TR1R2, containing sprT encoding SGT and the two positive regulatory genes sgtR1 and sgtR2, was investigated in various media. Cultivation in Ferm-0 gave 1.4 times more trypsin activity than in C5/L medium. In addition, replacement of 2% glucose and 1% skim milk in Ferm-0 with 2% dextrin and 1% tryptone (designated Ferm-II) enhanced trypsin activity 4.1-fold. To simplify the purification process, the supernatant from the S. griseus transformant cultured in Ferm-II medium was fractionated with ammonium sulfate (25-55%), then subjected to Hitrap Benzamidine FF affinity column chromatography. The specific activity of SGT purified by one-step chromatography was 69,550 unit/mg protein and the overall purification yield was above 8%, indicating that this method is more effective than those previously reported. Purified SGT was most active at pH 8.0 and $50^{\circ}C$, and it maintained activity between pH 7.0 and 9.0 and at temperatures up to $70^{\circ}C$. These enzymatic properties are very similar to those of authentic eukaryotic trypsin purified from bovine pancreas.

Recycling Technology of Waste Product in Electro Galvanizing Line of Steel Company

  • Lee, Jae-Young;Lee, H. H.;Kim, D. Y.;J. G. Sohn
    • Proceedings of the IEEK Conference
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    • 2001.10a
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    • pp.281-285
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    • 2001
  • This technology Provides an economical Production of high value added goods applicable to electro chemicals by recycling of waste products in EGL(Electro Galvanizing Line). The waste products produced in EGL contain potassium chloride (KCI), nickel and zinc. Highly pure KCI and Zinc Chloride which are raw material of electro plating, can be produced by the development of the recycling process. The scope of this study ranges from laboratory experiments to pilot test in plant. We have developed the whole process of recycling technology such as purification method of waste products, fabrication methods of electro chemicals, basic design of plant, pilot scale production and evaluation of pilot goods, Developed electro chemicals were pure enough to satisfy the specification of steel company.

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