اخبار و اعلانات

 آمار

تعداد نشریات283
تعداد نشریات سازمان81
تعداد شماره‌ها2,726
تعداد مقالات31,042
تعداد مشاهده مقاله27,944,668
تعداد دریافت فایل اصل مقاله16,240,575
Hazim Abdul Hameed, D, Hussein Ali, E. (1400). Extraction and Purification of Extracellular L-Glutamate Oxidase from Streptomyces. سامانه مدیریت نشریات علمی, 76(4), 769-779. doi: 10.22092/ari.2021.355928.1738
D Hazim Abdul Hameed; E Hussein Ali. "Extraction and Purification of Extracellular L-Glutamate Oxidase from Streptomyces". سامانه مدیریت نشریات علمی, 76, 4, 1400, 769-779. doi: 10.22092/ari.2021.355928.1738
Hazim Abdul Hameed, D, Hussein Ali, E. (1400). 'Extraction and Purification of Extracellular L-Glutamate Oxidase from Streptomyces', سامانه مدیریت نشریات علمی, 76(4), pp. 769-779. doi: 10.22092/ari.2021.355928.1738
Hazim Abdul Hameed, D, Hussein Ali, E. Extraction and Purification of Extracellular L-Glutamate Oxidase from Streptomyces. سامانه مدیریت نشریات علمی, 1400; 76(4): 769-779. doi: 10.22092/ari.2021.355928.1738

Extraction and Purification of Extracellular L-Glutamate Oxidase from Streptomyces

Archives of Razi Institute
مقاله 8، دوره 76، شماره 4، آذر و دی 2021، صفحه 769-779    اصل مقاله (927.87 K)
نوع مقاله: Original Articles
شناسه دیجیتال (DOI): 10.22092/ari.2021.355928.1738
نویسندگان
D Hazim Abdul Hameed* ؛ E Hussein Ali
Department of Biotechnology, Faculty of Applied Sciences, University of Technology, Iraq
چکیده
The bacterial isolates Streptomyces were obtained from the soil and cultivated in a wheat bran medium, which was used to produce the L–glutamate oxidase enzyme. The extracellular enzyme was then extracted using a cooling centrifugation process to obtain the filtrate that represents the crude enzyme. Afterward, the enzyme purification processes were carried out which included precipitation with ammonium sulfate as a preliminary purification step followed by dialysis to remove the salts. Next, ion-exchange chromatography and gel filtration were used to finish the purification process, and the enzyme activity was determined for each purification step. The results of purification of L-glutamate oxidase enzyme from streptomyces using ammonium sulfate showed that the specific activity was 8.25 units/mg protein with a saturation ratio of 60%. Moreover, the results of purification using a dialysis tube indicated that the specific activity was 9.5 units/mg protein. In addition, the result of purification using diethylaminoethyl cellulose ion column revealed that the specific activity was 25 unit/mg protein and the results of purification using gel filtration showed that the specific activity was 56 units/mg protein which was the best step in the purification process due to high specific activity of the enzyme. The optimum temperature and pH for the activity and stability of the enzyme were tested. Based on the findings, the optimum temperature for the activity of the enzyme was 37 °C. In addition, it was found that the optimum temperature range for the stability of the enzyme was 30-50 °C. Besides, the optimum pH for the activity was 7.0 and the optimum pH range for the enzyme stability was 5.0-7.0.
کلیدواژه‌ها
L-glutamate oxidase؛ L-glutamate؛ Streptomyces؛ Specific activity
سایر فایل های مرتبط با مقاله
مراجع
  1. Cooper J, Pritchard D. Biomolecular sensors for neurotransmitter determination: electrochemical immobilization of glutamate oxidase at microelectrodes in a poly (o-phenylenediamine) film. J Mater Sci Mater Electron. 1994;5(2):111-6.
  2. Valero E, Garcia-Carmona F. A Continuous Spectrophotometric Method Based on Enzymatic Cycling for Determiningl-Glutamate. Anal Biochem. 1998;259(2):265-71.
  3. Shi R, Stein K. Flow injection methods for determination of L-glutamate using glutamate decarboxylase and glutamate dehydrogenase reactors with spectrophotometric detection. Analyst. 1996;121(9):1305-9.
  4. Kusakabe H, Midorikawa Y, Fujishima T, Kuninaka A, Yoshino H. Purification and properties of a new enzyme, l-glutamate oxidase, from Streptomyces sp. X-119-6 grown on wheat bran. Agr Biol Chem. 1983;47(6):1323-8.
  5. Ling D, Wu G, Wang C, Wang F, Song G. The preparation and characterization of an immobilized L-glutamic decarboxylase and its application for determination of L-glutamic acid. Enzyme Microb Technol. 2000;27(7):516-21.
  6. Wachiratianchai S, Bhumiratana A, Udomsopagit S. Isolation, purification, and characterization of L-glutamate oxidase from Streptomyces sp. 18G. Electron J Biotechnol. 2004;7(3):09-10.
  7. Qingshan L, Lijun W, Yourong L. Color development with rational screening method for improved L-glutamate oxidase-producing strains. Enzyme Microb Technol. 1996;18(1):7-9.
  8. Sukhacheva M, Netrusov A. Streptomyces sp. Z-11-6, a novel producer of extracellular l-glutamate oxidase. Microbiology. 2000;69(1):13-6.
  9. Classics Lowry O, Rosebrough N, Farr A, Randall R. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951;193:265-75.
  10. Kuddus M, Joseph B, Ramteke PW. Production of laccase from newly isolated Pseudomonas putida and its application in bioremediation of synthetic dyes and industrial effluents. Biocatal Agric Biotechnol. 2013;2(4):333-8.
  11. Pointing S, Jones E, Vrijmoed L. Optimization of laccase production by Pycnoporus sanguineus in submerged liquid culture. Mycologia. 2000;92(1):139-44.
  12. Abood S, Hakeem I. Purification and characterization of salivary amylase inhibitor extracted from barley. Iraqi J Agric Sci. 2016;47(4).
  13. Al-easawi M. Extraction and Purification of Beta-Galactosidase from Local Almond and Its Use for Lactose Intolerance Treatment. Iraqi J Agric Sci. 2020;51(3):767-76.
  14. Whitaker JR, Bernhard RA. Experiments for: An Introduction to Enzymology. 1972.
  15. Erum S. Production of Peroxidases from Indigenous Fungal Strains and Their Application for Azo Dye Decolorization: Quaid-i-Azam University Islamabad, Pakistan; 2010.
  16. Younis R. Production, purification and inhibition of alginate ly-ase from local isolate of pseudomonas aeruginosa na11. Iraqi J Agric Sci. 2020;51(6):1726-39.
  17. Boeckx J, Hertog M, Geeraerd A, Nicolai B. Kinetic modelling: an integrated approach to analyze enzyme activity assays. Plant Methods. 2017;13(1):1-12.
  18. Noori H, Aziz G. Purification, Characterization, and Evaluation of Fibrinolytic Activity of Staphylokinase from Locally Isolated Staphylococcus aureus GH38. Iraqi J Agric Sci. 2020;51(4):1195-203.
  19. Daniel RM, Danson MJ, Eisenthal R, Lee CK, Peterson ME. The effect of temperature on enzyme activity: new insights and their implications. Extremophiles. 2008;12(1):51-9.
  20. Almeida VM, Marana SR. Optimum temperature may be a misleading parameter in enzyme characterization and application. PloS one. 2019;14(2):e0212977.
  21. Kusakabe H, Midorikawa Y, Kuninaka A, Yoshino H. Occurrence of a new enzyme, l-glutamate oxidase ina wheat bran culture extract of Streptomyces sp. X-119-6. Agr Biol Chem. 1983;47(1):179-82.
  22. Böhmer A, Müller A, Passarge M, Liebs P, Honeck H, Müller HG. A novel l‐glutamate oxidase from Streptomyces endus: Purification and properties. Eur J Biochem.1989;182(2):327-32.
  23. Whithaker J. Principles of enzymology for the food science. California: Marcel Dekker; 1994.
  24. Kennelly P, Rodwell V. Enzymes: Mechanism of action. 28th ed: Mc Graw Hill Lange; 2009. 51 p.
آمار
تعداد مشاهده مقاله: 1,049
تعداد دریافت فایل اصل مقاله: 431


counter
سامانه مدیریت نشریات علمی. قدرت گرفته از سیناوب