Antibacterial activity of the produced and purified L-glutamate oxidase from Streptomyces sp.

Document Type : Research Paper

Authors

College of Science for Women, University of Baghdad, Baghdad, Iraq

10.22124/cjes.2024.7503

Abstract

From Streptomyces species isolated from agricultural soil, an extracellular L-glutamate oxidase was recovered. The Streptomyces bacterial isolates were grown in a selective medium that had L-glutamate as a substrate. The filtrate, which is representative of the crude enzyme, was subsequently obtained by extracting the extracellular enzyme using a cooling centrifugation procedure. L-glutamate oxidase purification operations were then performed, starting with fractionation with 40% ammonium salts and continuing with ion-exchange chromatography and gel filtration with a final 8.25 units/mg and 61.8% yield. Both positive and negative bacteria were sensitive to the pure L-glutamate oxidase's effects. Purified L-glutamate oxidase outperformed other examined bacterial isolates in terms of activity, with MIC values of 32 µg/ml for Klebsiella pneumonia and 64 µg/ml for Staphylococcus aureus and Escherichia coli. Thus, pure L-glutamate oxidase may be viewed as a promising possibility in the rational design of new antibiotics.

Keywords


Abdul Hameed, H & Ali, H 2021, Extraction and purification of extracellular l-glutamate oxidase from Streptomyces. Archives of Razi Institute, 76: 769-779.
Ainur, S, Indira, R, Abaibek, Z, Krykbaevna, ZM, Shoshanovna, BA & Gulzhas, T 2023, Bacterial Mediation of Environmental Impacts: Unveiling novel pathways for mitigation and stewardship. Caspian Journal of Environmental Sciences, 21: 1171-1183.
Almashhadani, HA 2021, Synthesis of a CoO–ZnO nanocomposite and its study as a corrosion protection coating for stainless steel in saline solution. International Journal of Corrosion and Scale Inhibition, 10: 1294-306.
Almeida, VM & Marana, SR 2019, The optimum temperature may be a misleading parameter in enzyme characterization and application. PLOS ONE, 14: e0212977.
Barakate, M, Ouhdouch, Y, Oufdou, KH & Beaulieu, C 2018, Characterization of rhizospheric soil streptomycetes from Moroccan habitats and their antimicrobial activities. World Journal of Microbiology and Biotechnology, 18: 49-54.
Bradford, M 1976, A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding, Annal. Biochemistry, 72: 248-254.
Chater, KF 2016, Recent advances in understanding Streptomyces. F1000Research, 5: 16-21.
Chau, R, Kalaitzis, JA, Wood, SA & Neilan, BA 2018, Diversity and biosynthetic potential of culturable microbes associated with toxic marine animals. Marine Drugs, 11: 2695-2712, https://doi.org/10.3390/md11082695
Chaudhary, HS, Yadav, J, Shrivastava, AR, Singh, S, Singh, AK & Gopalan, N 2017, Antibacterial activity of actinomycetes isolated from different soil samples of Sheopur (A city in central India). Journal of Advanced Pharmaceutical Technology & Research, 4: 118-123.
Chowdhury, AP, Singh, RR, Bharadwaj, B, Bhuyana, MP & Jyoti, S 2019, Qualitative assessment on isolated tartar forming bacteria from dental caries by pattern of screening, assembling and antibiotic sensitivity. Journal of Research in Medical and Dental Science, 7: 140-157.
Hatif, ST, Al Taie, SF, Zeadan, AM, Awad Al-Nuaimi, ZJ & Rasheed, ZS 2023, Effect of Sidr honey on production of bacteriocin like inhibitory substances in some pathogenic bacteria. Caspian Journal of Environmental Sciences, 21: 1037-1041.
Kämpfer, P 2012, Family Streptomycetaceae. Volume 5: The Actinobacteria, In: MGW, Whitman, P, Kämpfer, H, Busse, M, Trujillo, W, Ludwig, K, Suzuki, A, Parte, (Eds.), Bergey’s Manual of Systematic Bacteriology Springer, New York, NY, USA.
Kusakabe, H, Midorikawa, Y, Kuninaka, A & Yoshino, H 2017, Occurrence of a new enzyme, l-glutamate oxidase in a wheat bran culture extract of Streptomyces sp. X-119-6. Agricultural and Biological Chemistry, 47: 179-182.
Li, QS & Zhong, JJ 2011, Short communication: Production of L-glutamate oxidase by Streptomyces sp. N1 in submerged fermentation. World Journal of Microbiology and Biotechnology, 12: 651-658.
Li, QS, Fang, QH, Zhu, JY & Zhong, JJ 2014, Hyperproduction of L-glutamate oxidase in submerged fermentation of Streptomyces sp. N1 with culture pH control and calcium addition. Applied Biochemistry and Biotechnology, 80: 97-106.
Maleki, H & Mashinchianm, O 2020, Characterization of Streptomyces Isolates with UV, FTIR Spectroscopy, and HPLC Analyses. BioImpacts, 1: 47-52.
Mohammed, AK, Al Shaheeb, S, Fawzi, OF, Almashhadani, HA & Kadhim, MM 2022, Evaluation of Interlukein-6 and Vitamin D in Patients with COVID-19. Research Journal of Biotechnology, 17.
Newitt, JT, Prudence, SMM, Hutchings, MI & Worsley, SF 2019, Biocontrol of cereal crop diseases using streptomycetes. Pathogens, 8: 78-82.
Oikawa, T, Watanabe, M, Makiura, H, Kusakabe, H, Yamade, K & Soda, K 2009, Production of d-glutamate from l-glutamate with glutamate racemase and l-glutamate oxidase. Bioscience, Biotechnology, and Biochemistrym, 63: 2168-2173.
Qingshan, L, Lijun, W & Yourong, L 2009, Colour development with rational screening method for improved L-glutamate oxidase-producing strains. Enzyme and Microbial Technology, 18: 7-9.
Quinn, GA, Banat, AM, Abdelhameed, AM & Banat, IM 2020, Streptomyces from traditional medicine: Sources of innovation in antibiotic discovery. Journal of Medical Microbiology, 69: 1040, 2020.
Rashad, FM, Fathy, HM, El Zayat, AS & Elghonaimy, AM 2016, Isolation and characterization of multifunctional Streptomyces species with antimicrobial, nematocidal, and phytohormone activities from marine environments in Egypt. Microbiological Research, 175: 34-47.
Seipke, RF, Kaltenpoth, M & Hutchings, MI 2020, Streptomyces as symbionts: An emerging and widespread theme? Fems. Microbiological Reviews, 36: 862-876.
Wachiratianchai, S, Bhumiratana, A & Udomsopagit, S 2014, Isolation, purification, and characterization of L-glutamate oxidase from Streptomyces sp. 18G. Electronic Journal of Biotechnology, 7: 09-10.