Effect of Vitreoscilla hemoglobin on production of a chemotherapeutic enzyme, L-asparaginase, by Pseudomonas aeruginosa.
The production of L-asparaginase, an enzyme widely used in cancer chemotherapy, is mainly regulated by carbon catabolite repression and oxygen. This study was carried out to understand how different carbon sources and Vitreoscilla hemoglobin (VHb) affect the production of this enzyme in Pseudomonas aeruginosa and its VHb-expressing recombinant strain (PaJC). Both strains grown with various carbon sources showed a distinct profile of the enzyme activity. Compared to no carbohydrate supplemented medium, glucose caused a slight repression of L-asparaginase in P. aeruginosa, while it stimulated it in the PaJC strain. Glucose, regarded as one of the inhibitory sugars for the production L-asparaginase by other bacteria, was determined to be the favorite carbon source compared to lactose, glycerol and mannitol. Furthermore, contrary to common knowledge of oxygen repression of L-asparaginase in other bacteria, oxygen uptake provided by VHb was determined to even stimulate the L-asparaginase synthesis by P. aeruginosa. This study, for the first time, shows that in P. aeruginosa utilizing a recombinant oxygen uptake system, VHb, L-asparaginase synthesis is stimulated by glucose and other carbohydrate sources compared to the host strain. It is concluded that carbon catabolite and oxygen repression of L-asparaginase in fermentative bacteria is not the case for a respiratory non-fermentative bacterium like P. aeruginosa.
Geckil
Hikmet
H
Department of Biology, Inonu University, Malatya 44280, Turkey. hgeckil@inonu.edu.tr
Gencer
Salih
S
Ates
Burhan
B
Ozer
Ufuk
U
Uckun
Mirac
M
Yilmaz
Ismet
I
D000970:Antineoplastic Agents / Q000302:isolation & purification / Q000378:metabolism; D001215:Asparaginase / Q000096:biosynthesis* / Q000302:isolation & purification / Q000627:therapeutic use; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D018929:Cell Culture Techniques / Q000379:methods*; D024861:Genetic Enhancement / Q000379:methods*; D005947:Glucose / Q000378:metabolism*; D006454:Hemoglobins / Q000235:genetics / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods; D011550:Pseudomonas aeruginosa / Q000145:classification / Q000201:enzymology* / Q000235:genetics; D013045:Species Specificity; D054793:Truncated Hemoglobins
D000970:Antineoplastic Agents; D001426:Bacterial Proteins; D006454:Hemoglobins; D054793:Truncated Hemoglobins; C082613:hemoglobin protein, Vitreoscilla; D001215:Asparaginase; D005947:Glucose