A novel glycosylated Cu/Zn-containing superoxide dismutase: production and potential therapeutic effect.
The fungal strain Humicola lutea 103 produces a naturally glycosylated Cu/Zn SOD. To improve its yield, the effect of an increased concentration of dissolved oxygen (DO) on growth and enzyme biosynthesis by the producer, cultivated in a 3 l bioreactor, was examined. Exposure to a 20% DO level caused a 1.7-fold increase of SOD activity compared to the DO-uncontrolled culture. Maximum enzyme productivity of SOD was approximately 300 x 10(3) U (kg wet biomass)(-1). The novel enzyme was purified to electrophoretic homogeneity. The presence of Cu and Zn were confirmed by atomic absorption spectrometry. The molecular mass of H. lutea Cu/Zn SOD was calculated to be 31870 Da for the whole molecule and 15936 Da for the structural subunits. The N-terminal sequence revealed a high degree of structural homology with Cu/Zn SOD from other prokaryotic and eukaryotic sources. H. lutea Cu/Zn SOD was used in an in vivo model for the demonstration of its protective effect against myeloid Graffi tumour in hamsters. Comparative studies revealed that the enzyme (i) elongated the latent time for tumour appearance, (ii) inhibited tumour growth in the early stage of tumour progression (73-75% at day 10) and (iii) increased the mean survival time of Graffi-tumour-bearing hamsters. Moreover, the fungal Cu/Zn SOD exhibited a strong protective effect on experimental influenza virus infection in mice. The survival rate increased markedly, the time of survival rose by 5.2 d and the protective index reached 86%. The H. lutea SOD protected mice from mortality more efficiently compared to the selective antiviral drug ribavirin and to commercial bovine SOD. In conclusion, our results suggest that appropriate use of the novel fungal SOD, applied as such or in combination with selective inhibitors, could outline a promising strategy for the treatment of myeloid Graffi tumour and influenza virus infection.
Angelova
Maria
M
Institute of Microbiology, Bulgarian Academy of Sciences, 26 Academician G. Bonchev, 1113 Sofia, Bulgaria1.
Dolashka-Angelova
Pavlina
P
Institute of Organic Chemistry, Bulgarian Academy of Sciences, 9 Academician G. Bonchev, 1113 Sofia, Bulgaria2.
Ivanova
Emilia
E
Institute of Microbiology, Bulgarian Academy of Sciences, 26 Academician G. Bonchev, 1113 Sofia, Bulgaria1.
Serkedjieva
Julia
J
Institute of Microbiology, Bulgarian Academy of Sciences, 26 Academician G. Bonchev, 1113 Sofia, Bulgaria1.
Slokoska
Ludmila
L
Institute of Microbiology, Bulgarian Academy of Sciences, 26 Academician G. Bonchev, 1113 Sofia, Bulgaria1.
Pashova
Svetlana
S
Institute of Microbiology, Bulgarian Academy of Sciences, 26 Academician G. Bonchev, 1113 Sofia, Bulgaria1.
Toshkova
Reneta
R
Institute of Experimental Pathology and Parasitology, Bulgarian Academy of Sciences, 23 Academician G. Bonchev, 1113 Sofia, Bulgaria3.
Vassilev
Spassen
S
Institute of Microbiology, Bulgarian Academy of Sciences, 26 Academician G. Bonchev, 1113 Sofia, Bulgaria1.
Simeonov
Ivan
I
Institute of Microbiology, Bulgarian Academy of Sciences, 26 Academician G. Bonchev, 1113 Sofia, Bulgaria1.
Hartmann
Hans-Jürgen
HJ
Anorganische Biochemie, Physiologisch-chemisches Institut der Universität, Tübingen, Hoppe-Seyler-Straße 4, D-72076, Tübingen, Germany4.
Stoeva
Stanka
S
Abteilung für Physikalische Biochemie des Physiologisch-chemischen Instituts der Universität, Tübingen, Hoppe-Seyler-Straße 4, D-72076, Tübingen, Germany5.
Weser
Ulrich
U
Anorganische Biochemie, Physiologisch-chemisches Institut der Universität, Tübingen, Hoppe-Seyler-Straße 4, D-72076, Tübingen, Germany4.
Voelter
Wolfgang
W
Abteilung für Physikalische Biochemie des Physiologisch-chemischen Instituts der Universität, Tübingen, Hoppe-Seyler-Straße 4, D-72076, Tübingen, Germany5.
D000595:Amino Acid Sequence; D000818:Animals; D019149:Bioreactors; D006224:Cricetinae; D005260:Female; D005658:Fungi / Q000201:enzymology* / Q000378:metabolism; D006031:Glycosylation; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism / Q000627:therapeutic use; D009052:Leukemia Virus, Murine; D007942:Leukemia, Experimental / Q000517:prevention & control; D008297:Male; D008647:Mesocricetus; D051379:Mice; D008813:Mice, Inbred ICR; D008969:Molecular Sequence Data; D009368:Neoplasm Transplantation; D009976:Orthomyxoviridae Infections / Q000188:drug therapy; D010100:Oxygen / Q000378:metabolism*; D012192:Retroviridae Infections / Q000517:prevention & control; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013482:Superoxide Dismutase / Q000302:isolation & purification / Q000378:metabolism* / Q000627:therapeutic use; D014412:Tumor Virus Infections / Q000517:prevention & control
D007527:Isoenzymes; D013482:Superoxide Dismutase; D010100:Oxygen