Isolation and Characterization of an L-Amino Acid Oxidase-Producing Marine Bacterium


One marine bacterial strain, R3, has been newly isolated from the intertidal zone of Dinghai sea area. Measurements of a-keto acids and H2O2existing in fermentation supernatant were carried out to show that R3 can produce L-amino acid oxidase (LAAO) with a broad substrate specificity. Physiological and biochemical analysis showed that it can grow great at the conditions with sodium chloride concentration of 1.5%-3%, temperature of 15?C -35?C and pH of 6-7. In addition, molecular identification of 16S rDNA was performed to show that R3 was proximal toPseudoalteromonasspp. with the highest identity of 98.5% toPseudoalteromonasrubra. Therefore, it was designated asPseudoalteromonassp. R3. Further studies are required to arrive at a better understanding of this LAAO and secure an application.

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Yu, Z. , Qiao, H. , Qiu, J. , Xu, P. and Li, P. (2013) Isolation and Characterization of an L-Amino Acid Oxidase-Producing Marine Bacterium. Engineering, 5, 376-380. doi: 10.4236/eng.2013.510B076.

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The authors declare no conflicts of interest.


[1] S. Torii, M. Naito and T. Tsuruo, “Apoxin I, a Novel Apoptosis-Inducing Factor with L-Amino Acid Oxidase Activity Purified from Western Diamondback Rattlesnake Venom,” Journal of Biological Chemistry, Vol. 272, No. 14, 1997, pp. 9539-9542.
[2] G. B. Naumanna, L. F. Silvaa, L. Silvaa, G. Faria, M. Richardson, K. Evangelista, et al., “Cytotoxicity and Inhibition of Platelet Aggregation Caused by an L-Amino Acid Oxidase from Bothrops leucurus Venom,” Biochimica et Biophysica Acta, Vol. 1810, No. 7, 2011, pp. 683-694.
[3] M. Y. Ahn, B. M. Lee and Y. S. Kim, “Characterization and Cytotoxicity of L-Amino Acid Oxidase from the Venom of King Cobra (Ophiophagus hannah),” The International Journal of Biochemistry & Cell Biology, Vol. 29, No. 6, 1997, pp. 911-919.
[4] R. G. Stábeli, S. Marcussi, G. B. Carlos, R. C. Pietro, H. S. Selistre-de-Araújo, J. R. Giglio, et al., “Platelet Aggregation and Antibacterial Effects of an L-Amino Acid Oxidase Purified from Bothrops alternatus Snake Venom,” Bioorganic & Medicinal Chemistry, Vol. 12, No. 11, 2004, pp. 2881-2886.
[5] Y. Abe, Y. Shimoyama, H. Munakata, J. Ito, N. Nagata and K. Ohtsuki, “Characterization of an Apoptosis-Inducing Factor in Habu Snake Venom as a Glycyrrhizin (GL)- Binding Protein Potently Inhibited by GL in Vitro,” Biological & Pharmaceutical Bulletin, Vol. 21, No. 9, 1998, pp. 924-927.
[6] D. H. Souza, L. M. Eugenio, J. E. Fletcher, M. S. Jiang, R. C. Garratt, G. Oliva, et al., “Isolation and Structural Characterization of a Cytotoxic L-Amino Acid Oxidase from Agkistrodon contortrix laticinctus Snake Venom: Preliminary Crystallographic Data,” Archives of Biochemitry and Biophysics, Vol. 368, No. 2, 1999, pp. 285-290.
[7] H. Takatsuka, Y. Sakurai, A. Yoshioka, T. Kokubo, Y. Usami, M. Suzuki, et al., “Molecular Characterization of L-Amino Acid Oxidase from Agkistrodon halys blomhoffii with Special Reference to Platelet Aggregation,” Biochimica et Biophysica Acta-Protein Structure and Molecular Enzymology, Vol. 1544, No. 1-2, 2001, pp. 267-277.
[8] S. A. Alib, S. Stoevab, A. Abbasi, J. M. Alam, R. Kayed, M. Faigle, et al. “Isolation, Structural, and Functional Characterization of an Apoptosis-Inducing L-Amino Acid Oxidase from Leaf-Nosed Viper (Eristocophis macmahoni) Snake Venom,” Archives of Biochemistry and Biophysics, Vol. 384, No. 2, 2000, pp. 216-226.
[9] P. Deolindo, A. S. Teixeira-Ferreira, R. A. DaMatta and E. W. Alves, “L-Amino Acid Oxidase Activity Present in Fractions of Bothrops jararaca Venom Is Responsible for the Induction of Programmed Cell Death in Trypanosoma cruzi,” Toxicon, Vol. 56, No. 6, 2010, pp. 944-955.
[10] B. G. Stilesa, F. W. Sextona and S. A. Weinstein, “Antibacterial Effects of Different Snake Venoms: Purification and Characterization of Antibacterial Proteins from Pseudechis australis (Australian King Brown or Mulga Snake) Venom,” Toxicon, Vol. 29, No. 9, 1991, pp. 1129-1141.
[11] Y. Hsiuchin, M. J. Paul, K. Ko-Chun, M. Kamio, M. W. Germann, C. D. Derby and P. C. Tai, “Cloning, Characterization and Expression of Escapin, a Broadly Antimicrobial FAD-Containing L-Amino Acid Oxidase from Ink of the Sea Hare Aplysia californica,” The Journal of Experi- mental Biology, Vol. 208, No. pt 18, 2005, pp. 3609-3622.
[12] M. A. Davis, M. C. Askin and M. J. Hynes, “Amino Acid Catabolism by an Area-Regulated Gene Encoding an L- Amino Acid Oxidase with Broad Substrate Specificity in Aspergillus nidulans,” Applied and Environmental Microbiology, Vol. 71, No. 7, 2005, pp. 3551-3555.
[13] A. Carbonnelle-Puscian, C. Copie-Bergman, M. Baia, N. Martin-Garcia, Y. Allory, C. Haioun, et al. “The Novel Immunosuppressive Enzyme IL4I1 Is Expressed by Neoplastic Cells of Several B-Cell Lymphomas and by Tumor-Associated Macrophages,” Leukemia, Vol. 23, No. 5, 2009, pp. 952-960.
[14] R. Genet, P. H. Benetti, A. Hammadi and A. Menez, “L- Tryptophan 2’,3’-Oxidase from Chromobacterium violaceum. Substrate Specificity and Mechanistic Implications,” The Journal of Biological Chemistry, Vol. 270, No. 40, 1995, pp. 23540-23545.
[15] R. L. Han-son, K. S. Bembenek, R. N. Patel and L. J. Szarka, “Transformation of N-Epsilon-CBZ-L-Lysine to CBT- L-Oxylysine Using L-Amino Acid Oxidase from Provi-dencia Alcalifaciens and L-2-Hydroxy-Iso-Caproate Dehydrogenase from Lactobacillus confuses,” Applied Microbiology and Biotechnology, Vol. 37, No. 5, 1992, pp. 599-603.
[16] M. Varadi, N. Adanyi, E. E. Szabo and N. Trummer, “Determination of the Ratio of D-and L-Amino Acids in Brewing by an Immobilize Amino Acid Oxidase Enzyme Reactor Coupled to Amperometric Detection,” Biosensors & Bioelectronics, Vol. 14, No. 3, 1999, pp. 335-340.
[17] M. Y. Ahn, K. S. Ryu, Y. W. Lee and Y. S. Kim, “Cytotoxicity and L-Amino Acid Oxidase Activity of Crude Insect Drugs,” Archives of Pharmacal Research, Vol. 23, No. 5, 2000, pp. 477-481.
[18] K. Ito, K. Hori and K. Miyazawa, “Purification and Some Properties of L-Amino Acid Oxidase from Amphiroa crassissima Yendo,” Hydrobiologia, Vol. 152, 1987, pp. 563-569.
[19] G. M. Brearley, C. P. Price, T. Atkinson and P. M. Hammond, “Purification and Partial Characterisation of a Broad-Range L-Amino Acid Oxidase from Bacillus carotarum 2Pfa Isolated from Soil,” Applied Microbiology and Biotechnology, Vol. 41, No. 6, 1994, pp. 670-676.
[20] L. Sikora and G. A. Marzluf, “Regulation of L-Amino Acid Oxidase and of D-Amino Acid Oxidase in Neurospora crassa,” Molecular & General Genetics, Vol. 186, No. 1, 1982, pp. 33-39.

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