تنقيه وتوصيف الاسبراجينيز المنتج من Bacillus sp.

محتوى المقالة الرئيسي

Sahar I. H.
زيد علي
Milad A.

الملخص

هدفت الدراسة الى عزل وتشخيص البكتيريا المنتجة للأسباراجينيز ، ثم تنقية الإنزيم وتوصيفه من أجل تحديد خصائصه في المستقبل. تم عزل 15 عزلة بكتيرية محلية من مواقع مختلفة في مدينة بغداد، وتم تشخيصها بوساطة الاختبارات المظهريه والكيميائية واختبارات الفيتيك، خضعت العزلات لعمليات الفحص الأولي لإنتاج الأسباراجينيز؛ حيث تم اختيار ثماني عزلات للفحص الثانوي ذات القابليه الاعلى في انتاج الهاله الصفراء في الوسط الصلب . بينت النتائج أن البكتريا العصويه من جنس Bacillus sp. لديها اعلى إنتاجية إنزيمية (7.5 وحدة / ملغم بروتين) بعد 24 ساعة من الحضانة ، حدتت إلى الظروف المثلى لإنتاج الانزيم بواسطة العزلة المختارة عن طريق التخمير المغمور ، حيث اختير الوسط (2) كأفضل وسيط للإنتاج وبوجود الفركتوز كأفضل مصدر للكربون عند درجة حموضة 6 ودرجه حرارة  40 درجة مئوية. تم تنقيه الانزيم بوساطه كروماتوغرافيا الترشيح الهلامي باستخدام جل السيفادكس –ج 150 ، حيث ازدادت عدد مرات التنقية النهائية بمقدار 2.5 مرة ، مما أدى إلى إنتاج إنزيم بنسبة 93.7٪. كما أظهر الانزيم المنقى أعلى فعاليه وثباتيه عند درجه حرارة 37 م ، و اعطى اعلى فعاليه عند الرقم الهيدروجيني 7.0 واعلى ثباتيه عند الرقم الهيدروجيني 8.0. و فقد الإنزيم نشاطه عند تعرضه لعدة أيونات معدنية بتركيزات 1 ، 5 ملي مولر.

تفاصيل المقالة

كيفية الاقتباس
1.
تنقيه وتوصيف الاسبراجينيز المنتج من Bacillus sp. Baghdad Sci.J [انترنت]. 5 ديسمبر، 2023 [وثق 20 مايو، 2024];20(6(Suppl.):2342. موجود في: https://bsj.uobaghdad.edu.iq/index.php/BSJ/article/view/9231
القسم
article
السير الشخصية للمؤلفين

زيد علي، قسم التقنيات الاحيائيه، كليه العلوم، جامعه بغداد، بغداد، العراق.

 

 

Milad A.، وزارة الصحه العراقيه، بغداد، العراق.

 

 

كيفية الاقتباس

1.
تنقيه وتوصيف الاسبراجينيز المنتج من Bacillus sp. Baghdad Sci.J [انترنت]. 5 ديسمبر، 2023 [وثق 20 مايو، 2024];20(6(Suppl.):2342. موجود في: https://bsj.uobaghdad.edu.iq/index.php/BSJ/article/view/9231

المراجع

Ofman MN, Krol M, Patel N, Krishnan S, Liu J, Saha V, et al. Rational engineering of l-asparaginase reveals importance of dual activity for cancer cell toxicity. Blood Journal. 2010; 117:1614-1621. https://doi.org/10.1182/blood-2010-07-298422.

Benchamin D, Sreejai R, Sujitha S and Albert C. Anti-proliferative activity of l-asparaginase enzyme from fungi on breast cancer. J. pharmacogn. phytochem. 2019; 8(1):407–410. https://www.phytojournal.com/7-6-468-716.pdf.

Gaufichon L, Rothstein SJ. and Suzuki A. Asparagine Metabolic Pathways in Arabidopsis. Plant Cell Physiol J. 2016; 57 (4): 675–689. https://doi.org/10.1093/pcp/pcv184.

Mohideen AKS. Molecular docking study of L-asparaginase I from Vibrio campbellii in the treatment of acute lymphoblastic leukemia (ALL). Eurobiotech J. 2020; 4 (1): 8-16. https://www.researchgate.net/publication/338682774_Molecular_docking_study_of_L-Asparaginase_I_from_Vibrio_campbellii_in_the_treatment_of_acute_lymphoblastic_leukemia_ALL.

Kaliwal Bb and Hosamani R. L-asparaginase an anti-tumor agent production by Fusarium equiseti using solid state fermentation. Int. J. Drug Discov. 2011; 3(2): 88-99. https://bioinfopublication.org/files/articles/3_2_2_IJDD.pdf

Ahmad N, Pandit NP and Maheshwari SK. L-asparaginase gene-a therapeutic approach towards drugs for cancer cell. Int. J. Biol. Sci. 2012; 2 (4): 1- 11. https://www.researchgate.net/publication/232261142_L-asparaginase_gene-_a_therapeutic_approach_towards_drugs_for_cancer_Cell

Verma N. L-asparaginase: a promising chemotherapeutic agent. Crit. Rev. Biotechnol. J. 2007; 27(1): 45-62. https://doi.org/10.1080/07388550601173926

Hymavathi M, Sathish T, Subba C, and Rao Prakasham RS. Enhancement of L-asparaginase production by isolated Bacillus circulans (MTCC 8574) using response surface methodology. Appl. Biochem. Biotechnol. J. 2009; 159: 191-8. https://doi.org/10.1007/s12010-008-8438-2

El-Naggar NEA and El-Shweihy NM. Bioprocess development for L-asparaginase production by Streptomyces rochei, purification and in-vitro efficacy against various human carcinoma cell lines. Sci. Rep. J. 2020; 10 (1):1-21. https://doi.org/10.1038/s41598-020-64052-x

Hussein SI, AL-Banaa AK, Noor HH and Mashkoor BA. Determination of the optimum conditions of laccase produced from local isolate of Citrobacter freundii using submerged fermentation. Asian Jr. of Microbiol. Biotech. Env. Sc. 2018; 20 (2): S181-S187. https://www.researchgate.net/publication/331498103_DETERMINATION_OF_THE_OPTIMUM_CONDITIONS_OF_LACCASE_PRODUCED_FROM_LOCAL_ISOLATE_OF_CITROBACTER_FREUNDII_USING_SUBMERGED_FERMENTATION

Hassan TJ, and Hussein SIa. Development of bioprocesses for production and purification of L- asparaginase from Staphylococcus aureus, and in vitro efficacy against human breast cell line. Iraqi J. Agric. Sci. 2023; 53(6):1525- 1538. https://doi.org/10.36103/ijas.v53i6.1668

Sharma A and Husain I. Optimization of medium components for extracellular glutaminase free asparaginase from Enterobacter cloacae. Int J Curr Microbiol. Appl. Sci. 2015; 4(1):296-309. https://www.ijcmas.com/vol-4-1/Anjana%20Sharma%20and%20Islam%20Husain.pdf.

Al-Dulimi AG, Al-Saffar AZ, Sulaiman GM, Khalil KA, Khashan KS, Al-Shmgani HS, et al. Immobilization of L-asparaginase on gold nanoparticles for novel drug delivery approach as anti-cancer agent against human breast carcinoma cells. J. Mater. Res. Tech.. 2020; 9(6): 15394-15411. https://doi.org/10.1016/j.jmrt.2020.10.021

Bradford M. A rapid and sensitive method for the quantitation of microgram quantities of protein using the principle of protein-dye binding. Anal. Biochem. J. 1976; 72: 248-254. https://doi.org/10.1006/abio.1976.9999

Dejong PJ. L-Asparaginase production by Streptomyces griseus. J. Appl. Microbiol.1972; 23(6):1163-1164. https://doi.org/10.1128/am.23.6.1163-1164.1972

Basha NS, Rekha R, Komala M and Ruby S. Production of extracellular anti-leukaemic enzyme L-asparaginase from marine actinomycetes by solid state and submerged fermentation: Purification and characterization. Trop. J. Pharm. Res. 2009; 8(4): 353-360. https://www.researchgate.net/publication/43561132_Production_of_Extracellular_Anti-leukaemic_Enzyme_Lasparaginase_from_Marine_Actinomycetes_by_Solidstate_and_Submerged_Fermentation_Purification_and_Characterisation.

Aziz GM, Hussein SI, Abbass SD, Ibrahim AL and Abbas DK. Degradation of reactive dyes using immobilized peroxidase purified from Nigella sativa. Iraqi J. Agric. Sci. 2021; 52(6):1365-1374. https://doi.org/10.36103/ijas.v52i6.1476

Hanaa NS, and Hussein SI. Assessment of purified collagenase inhibition activity from Staphylococcus aureus HN77 by some local plants extract. Iraqi J. Agric. Sci. 2022; 53(5):1035- 1047. https://doi.org/10.36103/ijas.v53i5.1617

Devi AL and Ramanjaneyulu R. Isolation of L-Asparaginase producing microbial strains from soil samples of telangana and andhra Pradesh States. Int J Curr Microbiol Appl Sci. 2016; 5(10): 1105-1113. https://www.ijcmas.com/5-10-2016/A.S.%20Lalitha%20Devi%20and%20R.%20Ramanjaneyulu.pdf.

Wakil SS and Adelegan AA. Screening, production and optimization of L-asparaginase from soil bacteria isolated in Ibadan, South-western Nigeria. Basic Appl. Sci. J. 2015; 11: 39-51. https://www.researchgate.net/publication/272087886_Screening_Production_and_Optimization_of_L-Asparaginase_From_Soil_Bacteria_Isolated_in_Ibadan_South-western_Nigeria

Hughes SG. Variation in enzyme production as a function of the physiological state of bacteria. A thesis presented for the degree of Master of Science Department of Molecular Biology University of Edinburgh. 1971. https://era.ed.ac.uk/bitstream/handle/1842/15054/Hughes1971.Pdf;sequence=1

Poongothai E, Siddharthan N, Hemalatha N and Balagurunathan R. Production and Partial Purification of L-Asparaginase Enzyme from Bacteria. IAJPS. 2017; 4(12): 4798-4803. https://doi.org/10.5281/zenodo.1134384

El-Naggar NEA and El-Shweihy NM. Bioprocess development for L-asparaginase production by Streptomyces rochei, purification and in-vitro efficacy against various human carcinoma cell lines. Sci. Rep. J. 2020; 10 (1):1-21. https://doi.org/10.1038/s41598-020-64052-x

Bhargavi PL, Kumar BS and Prakasham SR. Impact of nutritional factors verses biomass and serralysin production in isolated Serratia marcescens. Curr. trends. biotechnol. Pharm. J. 2012; 6(4): 449-457. https://www.researchgate.net/publication/287750130_Impact_of_nutritional_factors_verses_biomass_and_serralysin_production_in_isolated_Serratia_marcescens

Moawad W, El-Naby MA, Darwish DB. and Sherief AA. Production, optimization and characterization of L-asparaginase from Aspargillus flavus. Mansoura Journal of Biology. 2013; 38 (2). https://www.researchgate.net/publication/349731162_Mansoura_University_Faculty_of_Science_PRODUCTION_OPTIMIZATION_AND_CHARACTERIZATION_OF_LASPARGINASE_FROM_ASPERGILLUS_FLAVUS_PRODUCTION_OPTIMIZATION_AND_CHARACTERIZATION_OF_LASPARGINASE_FROM_ASPERGIL

Tortora GJ, Funke BR and Case CL. Microbiology. 8th ed. Pearson Education, Inc. San Francisco. New York. 2004. https://books.google.iq/books/about/Microbiology.html?id=q55ruAAACAAJ&redir_esc=y

Hussein SI, Hayder NH and Aziz GM. Biodegradation of Some Hydrocarbon Compounds by Free and Immobilized Laccase Produced from Local Isolates of Pseudomonas aeruginosa using a bioreactor. A thesis presented for the degree of doctor of philosophy in biotechnology department. Baghdad University. Iraq. 2017. https://www.researchgate.net/publication/369230060_Biodegradation_of_Some_Hydrocarbon_Compounds_by_Free_and_Immobilized_Laccase_Produced_from_Local_Isolates_of_Pseudomonas_aeruginosa_using_a_bioreactor

Bull AT and Bushnel ME. Environmental control of fungal growth In: The filamentous fungi. (Eds. Smith, J.E. and Berry, D.E.). 1976; (2):1-26. Edward Arnold, London.

Lazazzera BA. Quorum sensing and starvation. Signals for entry into the stationary phase. Curr. Microbiol. J. 2000; 3: 177 182. https://doi.org/10.1016/S1369-5274(00)00072-2

Narayana KJP, Kumar KG and Vijayalakshmi M. L-asparaginase production by Streptomyces albidoflavus. Indian J. Microbiol. 2008; 48:331-336. https://doi.org/10.1007/s12088-008-0018-1.

Dhevagi P, and Poorani E. Isolation and characterization of L-asparaginase from marine actinomycetes. Indian J. Biotechnol. 2006; 5: 514-520. https://www.researchgate.net/publication/266339260_Isolation_and_characterization_of_L-asparaginase_from_marine_actinomycetes

Chauhan A, Mohindra A and Prabha V. Purification and characterization of collagenase from Bacillus altitudinis. J. Med. Robot. Res. 2020; 45: 2395.6623.6120.1. https://www.researchgate.net/publication/363535370_Purification_and_characterization_of_collagenase_from_Bacillus_altitudinis

Siddalingeshwara KG and Lingappa K. Production and characterization of L-asparaginase-A Tumour inhibitor. Int. J. Pharmtech Res. 2011; 3(1):314-319. https://www.semanticscholar.org/paper/Production-and-Characterization-of-L-Asparaginase-A-K.G/0ac723cf5e5b6e207307fcce6ec2bf56cc64d596

Meghavarnam KA and SavithaJ. Purification and characterization of therapeutic enzyme L-asparaginase from a tropical Soil fungal isolate Fusarium culmorum ASP-87. MOJPB. J. 2015; 2(6): 00064. https://www.researchgate.net/publication/297871468_Purification_and_Characterization_of_Therapeutic_Enzyme_LAsparaginase_from_a_Tropical_Soil_Fungal_Isolate_Fusarium_Culmorum_ASP-87

Clive DA. Guide to protein isolation. Kluwer academic publisher. New York. 2002. https://link.springer.com/book/10.1007/978-94-017-0269-0

Moorthy V, Ramalingam A, Sumantha A and Shankaranaya RT. Production, purification and characterization of extracellular L-asparaginase from a soil isolate of Bacillus sp. Afr. J. Microbiol Res. 2010; 4(18):1862-1867. https://www.researchgate.net/publication/228616101_Production_purification_and_characterisation_of_extracellular_L-asparaginase_from_a_soil_isolate_of_Bacillus_sp

Hassan TJ and Hussein SIb .Evaluation of Anticancer Activity of Partially Purified L-Asparaginase Produced by Staphylococcus aureus. A Thesis presented for the degree of master in biotechnology department. Baghdad University. Iraq. 2023. https://www.researchgate.net/publication/373014242_Evaluation_of_Anticancer_Activity_of_Partially_Purified_LAsparaginase_Produced_by_Staphylococcus_aureus

Segel JJ. Biochemical Calculation. John Wiley and sons. 1976. https://books.google.iq/books/about/Biochemical_Calculations.html?id=XxeUEAAAQBAJ&redir_esc=y

Ohta Y, Nogi Y, Miyazaki M, Li Z, Hatada Y, Ito S and Horikoshi K. Enzymatic properties, nucleotide and amino acid sequence of a theromstable beta-agarase from the novel marine isolate. JAMB-A94. J. Biosci Biotech and Biochem. 2004; 68(5):1073-1081. https://doi.org/10.1007/s00253-004-1573-y.

Mohindra A, Chauhan A and Prabha V. Purification and characterization of collagenase from Bacillus altitudinis. J. Med. Robot Research. 2020;45: 2395.6623.6120.1. https://www.researchgate.net/publication/363535370_Purification_and_characterization_of_collagenase_from_Bacillus_altitudinis.

41. Barker SA and Shirley JA. Microbial enzyme and bioconversion. In: Economic Microbiology. (ed. Rose, A.H.). 5: 173–186. Academic Press. London. 1980. https://link.springer.com/article/10.1007/BF02877133

المؤلفات المشابهة

يمكنك أيضاً إبدأ بحثاً متقدماً عن المشابهات لهذا المؤلَّف.