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Purification and characterization of L-Amino acid oxidase from Aspergillus terreus MZ769058

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Abstract

L-Amino acid oxidase has significant values in different biotechnology sectors. In this study, Aspergillus terreus MZ769058 was reported as new fungal isolate for production of this enzyme. It was purified with high degree of purification fold (2.55) and high specific activity (132.5 U/mg protein). Electrophoresis confirmed its homodimer nature with molecular weight of whole enzyme as 180 kDa and its subunit as 90 kDa. This enzyme showed maximum activity of 193.5 U/l at optimum value of pH 6.0. The enzyme was active throughout a wide range of temperatures and showed maximum activity (227.08 U/l) at optimum temperature 30 °C. The value of Michaelis parameters of Km and Vmax was estimated as 26 mM and 250 µmole/min/mg proteins, respectively. The catalytic efficiency of this enzyme (Kcat) value was determined as 2.5 µmole/min/mg. Metal salts like FeSO4 (85.4%), Na2MO4 (81.2%), and CuSO4 had showed negative effect on its activity. This enzyme was strongly inhibited with α-napthol (34.4%), EDTA (34.2%), Glycine (39%) sodium azide (41.4%), and riboflavin (85.3%). Fourier transform infrared spectroscopy had confirmed the presence of the amine and aldehyde groups with C-H stretch, C = O stretch, C–O stretch at peak of 2927.95, 1745.25, and 1078.64 cm−1. This enzyme showed greater zone of inhibition against growth of Bacillus subtillis ATCC 11774 (6 ± 0.14) and Staphylococcus aureus ATCC 25923 (12.00 ± 0.12). This enzyme could be used for effective therapeutic agent in biotechnological sector.

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References

  • Bohmer A, Muller A, Passarge M, Liebs P, Honeck H, Muller HG (1989) A novel l-glutamate oxidase from Streptomyces endus: Purification and properties. Eur J Biochem 182(2):327–332

    Article  CAS  PubMed  Google Scholar 

  • Brumfitt W, Hamilton-Miller JM, Franklin I (1990) Antibiotic activity of natural products: 1. Propolis. Microbios 62(250):19–22

    CAS  PubMed  Google Scholar 

  • Burgess RR (2009) Protein precipitation techniques. Methods Enzymol 463:331–342

    Article  CAS  PubMed  Google Scholar 

  • Chauhan N, Tyagi AK, Kumar P, Malik A (2016) Antibacterial potential of Jatropha curcas synthesized silver nanoparticles against food borne pathogens. Front Microbiol 7:1748

    Article  PubMed  PubMed Central  Google Scholar 

  • Choukade R, Jaiswal A, Kango N (2020) Characterization of biogenically synthesized silver nanoparticles for therapeutic applications and enzyme nanocomplex generation. 3 Biotech 10(11):1–13

    Article  Google Scholar 

  • Elamawi RM, Al-Harbi RE, Hendi AA (2018) Biosynthesis and characterization of silver nanoparticles using Trichoderma longibrachiatum and their effect on phytopathogenic fungi. Egypt J Biol Pest Control 28(1):1–11

    Article  Google Scholar 

  • El-Sayed AS, Shindia AA, Zaher Y (2012) L-Amino acid oxidase from filamentous fungi: screening and optimization. Ann Microbiol 62(2):773–784

    Article  CAS  Google Scholar 

  • El-Sayed AS, Shindia AA, Zaher YA (2013) Purification and characterization of L-amino acid oxidase from the solid-state grown cultures of Aspergillus oryzae ASH. Microbiology 82(6):762–771

    Article  CAS  Google Scholar 

  • Geueke B, Hummel W (2002) A new bacterial L-amino acid oxidase with broad substrate specificity: purification and characterization. Enzyme Microb Technol 31(1–2):77–87

    Article  CAS  Google Scholar 

  • Gogichaeva NV, Alterman MA (2019) Amino acid analysis by means of MALDI TOF mass spectrometry or MALDI TOF/TOF tandem mass spectrometry. Methods Mol Biol (clifton, N.J.) 2030:17–31. https://doi.org/10.1007/978-1-4939-9639-1_3

    Article  CAS  Google Scholar 

  • Hiu JJ, Yap MKK (2020) Cytotoxicity of snake venom enzymatic toxins: phospholipase A2 and l-amino acid oxidase. Biochem Soc Trans 48(2):719–731

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227(5259):680–685

    Article  CAS  PubMed  Google Scholar 

  • Li G, He D, Qian Y, Guan B, Gao S, Cui Y, Wang L (2011) Fungus-mediated green synthesis of silver nanoparticles using Aspergillus terreus. Int J Mol Sci 13(1):466–476

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lowry OH (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

    Article  CAS  PubMed  Google Scholar 

  • Neema KN, Vivek HK, Nanjunda S, Kumar JR, Priya BS (2015) Purification and biochemical characterization of L-amino acid oxidase from western region Indian cobra (Naja naja) venom. Int J Pharm Pharm Sci 7:167–171

    CAS  Google Scholar 

  • Oike K, Groger H (2020) Process properties of an l-amino acid oxidase from Hebeloma cylindrosporum for the synthesis of phenylpyruvic acid from l-phenylalanine. J Biotechnol 323:203–207

    Article  CAS  PubMed  Google Scholar 

  • Singh S, Gogoi BK, Bezbaruah RL (2014) Aspergillus fumigatus. L-amino acid oxidase-two step purification and characterization of the enzyme. J MicrobBiochem Technol 6:096–101

    Google Scholar 

  • Souza DH, Eugenio LM, Fletcher JE, Jiang MS, Garratt RC, Oliva G, Selistre-de-Araujo HS (1999) Isolation and structural characterization of a cytotoxic L-amino acid oxidase from Agkistrodon contortrix laticinctus snake venom: preliminary crystallographic data. Arch Biochem Biophys 368(2):285–290

    Article  CAS  PubMed  Google Scholar 

  • Stabeli RG, Marcussi S, Carlos GB, Pietro RC, Selistre-de-Araujo HS, Giglio JR, Soares AM (2004) Platelet aggregation and antibacterial effects of an L-amino acid oxidase purified from Bothrops alternatus snake venom. Bioorg Med Chem 12(11):2881–2886

    Article  CAS  PubMed  Google Scholar 

  • Tyagi S, Tyagi PK, Gola D, Chauhan N, Bharti RK (2019) Extracellular synthesis of silver nanoparticles using entomopathogenic fungus: characterization and antibacterial potential. SN Applied Sciences 1(12):1–9

    Article  Google Scholar 

  • Van Boven A, Tan PST, Konings WN (1988) Purification and characterization of a dipeptidase from Streptococcus cremoris Wg2. Appl Environ Microbiol 54(1):43–49

    Article  PubMed  PubMed Central  Google Scholar 

  • Wellner D, Meister A (1960) Evidence for an intermediate in the oxidation of reduced L-amino acid oxidase by molecular oxygen. J Biol Chem 235:PC12–PC13

    Article  CAS  PubMed  Google Scholar 

  • Yadav M, Singh P (2023) Production of L-amino acid oxidase from new fungal isolate Aspergillus terreus MZ769058 and optimization of their immobilization parameters. Vegetos 36:851–863. https://doi.org/10.1007/s42535-022-00457-5

    Article  Google Scholar 

  • Yu Z, Qiao H (2012) Advances in non-snake venom L-amino acid oxidase. Appl Biochem Biotechnol 167(1):1–13

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

The authors are grateful to the Department of Biosciences and Biotechnology, Banasthali Vidyapith, for providing infrastructural facilities and moral support for completing the research work successfully.

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Authors and Affiliations

  1. Department of Bioscience and Biotechnology, Banasthali Vidyapith, Tonk, Rajasthan, India

    Monika Yadav

  2. Institute of Allied Medical Science and Technology, NIMS University Rajasthan, Jaipur, Rajasthan, India

    Shefali Agarwal, Sushant Agarwal & Priyanka Singh

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  1. Monika Yadav
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  2. Shefali Agarwal
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  4. Priyanka Singh
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Correspondence to Priyanka Singh.

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Yadav, M., Agarwal, S., Agarwal, S. et al. Purification and characterization of L-Amino acid oxidase from Aspergillus terreus MZ769058. Vegetos (2024). https://doi.org/10.1007/s42535-024-00962-9

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