Pseudomonas aeruginosa Expressing Vitreoscilla Hemoglobin Shows Increased Production of L-Lysine α-Oxidase: an Enzyme used in Cancer Therapy

Year 2018, Volume: 13 Issue: 2, 47 - 52, 19.09.2018

Hüseyin Kahraman

Abstract

L-lysine α-oxidase
(LO) is one of a few microbial enzymes with therapeutic potential in certain
cancers. The enzyme has been determined in several bacteria and fungi. Its
production is mainly regulated by carbon sources and oxygen. . Thus, the aim of
this study was to determine the nutritional requirements and effect of oxygen
concentration on the production of LO in P. aeruginosa, and in their
recombinants using a highly efficient oxygen uptake system, the Vitreoscilla
hemoglobin. This study concerns the effect of a higher oxygen uptake provided
by a recombinant system, the Vitreoscilla
hemoglobin, on the production of LO in Pseudomonas
aeruginosa. The results showed that
the recombinant bacterium expressing Vitreoscilla
hemoglobin gene (vgb) had distinct
L-lysine activity from the host strain under both carbon catabolite repression
and no repression conditions. In a rich medium supplemented with glucose, the
recombinant strain showed 20-40 % higher L-lysine activity than the host
strain. This difference was even more significant in the medium with no glucose
supplement, where the recombinant strain showed almost 2-fold higher enzyme
activity throughout the incubation. The results sometime were contradictory in
terms of the effect of carbon source (mainly glucose) and oxygen on the
production of this enzyme.

Keywords

Vitreoscilla hemoglobin, Psedomonas aeruginosa, L-lysine α-oxidase

References

• 1. Saurina, J., Herna’ndez-Cassou, S., Alegret, S., and Fa’bregas, E. (1999). Determination of lysine in pharmaceutical samples containing endogenous ammonium ions by using a lysine oxidase biosensor based on an all-solid-state potentiometric ammonium electrode. Biosens. Bioelect, 14: 67-75. 2. Saurina J., Herna´ndez-Cassou S., Alegret S., and Fa`bregas E. (1999). Amperometric determination of lysine using a lysine oxidase biosensor based on rigid-conducting composites. Biosens. Bioelect., 14: 211-220. 3. Treshalina, H.M., Lukasheva, E.V., Sedakova, L.A., Firsova, G.A., Guerassimova, G.K., Gogichaeva, N.V., and Berezov, T.T. (2000). anticancer enzyme L-lysine α-oxidase. Appl. Biochem. Biotechnol, 88: 267-273. 4. Kusakabe, H., Kodama, K., Kuninaka, A., Yoshino, H., Misono, H., and Soda, K. (1980). A new antitumor enzyme, L-lysine α-oxidase from Trichoderma viride, Purification and enzymological properties. J. Biol. Chem., 255: 976-981. 5. Galaris, D., and Evangelou, A. (2002). The role of oxidative stress in mechanisms of metal-induced carcinogenesis. Crit. Rev. Oncol. Hematol., 42: 93-103. 6. Li, W., Nugent, M. A., Zhao, Y., Chau, A. N., Li, S. J., Chou, I., Liu, G., and Kagan, H. M. (2003). Lysyl oxidase oxidizes basic fibroblast growth factor and inactivates its mitogenic potential. J. Cell. Biochem., 88: 152-164. 7. Lukasheva, E.V., and Berezov, T.T. (2002). L-Lysine-α-Oxidase: Physicochemical and biological properties. Biochem., 67: 1152-1158. 8. Murthy, S. N., and Janardanasarma, M. K. (1999). Identification of L-amino acid/ L-lysine α-amino oxidase in mouse brain. Molec. Cell. Biochem., 197: 13-23. 9. Almuaibed, A. M., and Townshend, A. (1997). Flow injection amperometric and chemiluminescence individual and simultaneous determination of lysine and glucose with immobilized lysine oxidase and glucose oxidase. Anal. Chim. Acta, 338: 149-154. 10. Geckil, H., Gencer, S., and Uckun, M. (2004). Vitreoscilla hemoglobin expressing Enterobacter differently to carbon catabolite and oxygen repression for production of L-asparaginase, an enzyme used in cancer therapy. Enzy. Microb. Technol., 35: 182-189. 11. Chung, J. W., Webster, D. A., Pagilla, K. R., and Stark, B. C. (2001). Chromosomal integration of the Vitreoscilla hemoglobin gene in Burkholderia and Pseudomonas for the purpose of producing stable engineered strains with enhanced bioremediating ability. J. Indust. Microbiol.Biotechnol., 27: 27-33. 12. Mukherjee, J., Majumdar, S., and Scheper, T. (2000). Studies on nutritional and oxygen requirements for production of L-asparaginase by Enterobacter aerogenes. Appl. Microbiol. Biotech., 53: 180-184. 13. Saurina, J., Herna’ndez-Cassou, S., Fa’bregas, E., and Alegret, S. (1998). Potentiometric biosensor for lysine analysis based on a chemically immobilized lysine oxidase membrane. Analy. Chim. Acta, 371: 49-56. 14. Lowry, O. H., Rosebrough, N. J., Farr, A. L., and Randall, R. J. (1951). Protein measurement with the folin phenol reagent. J. Biol. Chem., 193: 265-275. 15. Geckil, H., Gencer, S., Kahraman, H., and Erenler, S.O. (2003). Genetic engineering of Enterobacter aerogenes with Vitreoscilla hemoglobin gene: cell growth, survival and antioxidant enzyme status under oxidative stress. Res. Microbiol., 154: 425-431.