Biomedical subjects

G Ozcengiz

Publications and source records attributed to G Ozcengiz.

11 recordsLinked to original sources

PubMed2005

Inorganic phosphate has a crucial effect on Cry3Aa delta-endotoxin production.

AIMS: The study aimed at increasing Cry3Aa delta-endotoxin production by a local isolate of Bacillus thuringiensis (B.t. strain Mm2). To this end, different nutritional conditions were tested for their effects on Cry3Aa yields. METHODS AND RESULTS: Bacillus thuringiensis Mm2 was grown by shaking at 30 degrees C in different media. Samples were taken from the cultures at intervals and used for protein extraction. SDS-PAGE was performed for toxin analysis. Inclusion of inorganic phosphate (Pi) into the Difco's sporulation medium at an increased level of 200 mmol l-1 caused a fivefold increase (from 3 to 15.6 microg ml-1) in toxin production. Omission of FeSO4 from the medium decreased this yield by half. Resuspension experiments suggested catabolite repression of toxin biosynthesis by glucose. The inclusion of high Pi invariably increased toxin synthesis, even in the absence of sugars. CONCLUSIONS: Inorganic phosphate had the most striking effect on toxin biosynthesis. Iron effect was found to be unique to our isolate whereas Pi effect seemed to be common to the biosynthesis of Cry3Aa-type toxins. Stimulation of toxin synthesis by Pi did not seem to represent a relief from glucose repression. SIGNIFICANCE AND IMPACT OF THE STUDY: Bacillus thuringiensis is the most versatile biopesticide for use in pest management. Regarding cost-effectiveness of related fermentations, high Pi supplement drastically increases Coleoptera-specific toxin synthesis.

Animals↗

PubMed2003

Colicinogeny in local isolates of salmonellae and plasmid transfer studies.

Colicinogeny was determined in local isolates of S. typhimurium and S. enteritidis. Fourteen out of 35 S. typhimurium isolates of hospital origin were colicin producers whereas only one chicken isolate out of 82 S. enteritidis isolates of various origin (human, chicken or egg) produced colicin. A colicin producing, cephalothin (Cpt)- and piperacillin (Prl)-resistant local isolate of Salmonella havana (H32) harbored 4 plasmids of 54.0, 28.4, 2.7 and 1.9 kb. Upon curing its plasmids, the strain lost the ability to produce colicin and resistance to antibiotics and no longer expressed smooth lipopolysaccharide (LPS). The outer membrane protein (OMP) of 34.6 kDa was also lost. Using nalidixic acid (Nal)-resistant mutant of the cured strain in conjugation experiments, 10 out of 27 transconjugants were found to be resistant to Nal and Prl, 10 were resistant to Nal and Cpt and 7 showed Nal, Prl and Cpt resistance. Cpt and Prl resistance were determined by 54.0 and 28.4 kb plasmids, respectively. There was no direct correlation between plasmid contents and colicinogeny, LPS and OMP profiles.

Animals↗

PubMed2001

Tn10 insertional mutations of Bacillus subtilis that block the biosynthesis of bacilysin.

Transposon mutagenesis was employed to isolate the gene(s) related with the biosynthesis of dipeptide antibiotic in Bacillus subtilis PY79 (a prototrophic derivative of the standard 168 strain). The blocked mutants were phenotypically selected from the transposon library by bioassay and the complete loss of biosynthetic ability was verified through ESI-mass spectrometry analysis. Four different bacilysin nonproducer mutants (Bac(-)::Tn10(ori-spc)) were isolated from the transposon library. The genes involved in bacilysin biosynthesis were identified as thyA (thymidilate synthetase), ybgG (unknown; similar to homocysteine methyl transferase) and oppA (oligopeptide permease), respectively. The other blocked gene was yvgW (unknown; similar to heavy metal-transporting ATPase); however, backcross studies did not verify its involvement in bacilysin biosynthesis. This gene, on the other hand, appeared to be necessary for efficient sporulation and transformation. Opp involvement was significant as it suggested that bacilysin biosynthesis is under or a component of the quorum sensing pathway which has been shown to be responsible for the establishment of sporulation, competence development and onset of surfactin biosynthesis. For verification, it was necessary to check the involvement of peptide pheromones (PhrA or PhrC) internalized by the Opp system and response regulator ComA as the essential components of this global control. phrA, phrC and comA deleted mutants of PY79 were thus constructed and the latter two genes were shown to be essential for bacilysin biosynthesis.

ATP-Binding Cassette Transporters↗

PubMed1999

Immunoadjuvant action of plasmid DNA in liposomes.

Bacterial DNA and oligodeoxynucleotides containing immunostimulatory sequences with a CpG motif stimulated a Th1 type response in vivo. The adjuvant action of a non-coding plasmid DNA derived from pRc/CMV HBS (encoding the S region of hepatitis B surface antigen, HBsAg) in mice was investigated. The role of methylation on the adjuvanticity of the plasmid as well as the effect of vaccine formulation employed on the outcome of antigen-specific humoral and cellular responses were also studied. The results demonstrated that plasmid DNA acted as a Th1 promoting adjuvant when mixed as such or co-entrapped in liposomes with a very low dose of antigen. However, the adjuvant activity was lost when separate liposome entrapped formulations of both the antigen and the plasmid DNA were mixed, indicating a necessity for the antigen and the plasmid DNA to contact the same APC for optimal immune activation. A decreased adjuvanticity of plasmid DNA upon methylation with HpaII methyltransferase was also demonstrated. A mechanism that may help partially explain the reduction in adjuvanticity after modification of C residues is also discussed.

Adjuvants, Immunologic↗

PubMed1999

Assignment of biological functions to specific plasmids in a local isolate of Rhizobium cicer.

The genetic basis of symbiosis, mucoid structure and melanin biosynthesis in a local isolate of Rhizobium cicer was investigated. The strain was a very effective symbiont and produced substantial amounts of exopolysaccharides and melanin. It harbored three high-molar-mass plasmids of 55, 80 and 130 kb, respectively. Thirty-one melanin production-negative (Mep-) derivatives were obtained through plasmid curing. The plasmid patterns of cured derivatives indicated that the loss of 55 and/or 80 kb plasmids was sufficient for obtaining Mep- phenotype. The specific involvement of 55-kb plasmid in melanin production was confirmed upon re-introduction of this plasmid into one of the Mep- derivatives. Further investigation also indicated that the 130-kb plasmid might be necessary for both mucoidal appearance and symbiotic functions.

Anti-Bacterial Agents↗

PubMed1998

Multiple drug resistance patterns and plasmid profiles of non-typhi salmonellae in Turkey.

A total of 259 clinical isolates of nonrepetitive non-typhi salmonellae (NTS) were examined for antibiotic resistance patterns and plasmid content. The antibiotics used were amoxicillin-clavulanic acid (AMC), ampicillin (AM), aztreonam (ATM), carbenicillin (CB), cefixime (CFM), cefotaxime (CTX), cefoxitin (FOX), ceftazidime (CAZ), ceftriaxone (CRO), chloramphenicol (C), ciprofloxacin (CIP), gentamicin (GM), imipenem (IPM), ofloxacin (OFX), tetracycline (TE), trimethoprim-sulfomethoxazole (SXT). Multi-drug resistant (MDR) strains comprised 19.3% of the total isolates (50/259) and almost all were S. typhimurium (49/50). Fifteen different patterns of resistance was observed, AM/CB/C/AMC/TE and AM/CB/C/AMC/SXT/GM/CTX/CRO/CAZ/CFM/ATM being the most frequent patterns. Twenty-eight out of 50 multiresistant isolates were found to contain at least one plasmid (mean five) and the size of the plasmids ranged between 1.7 and 158 kb. Plasmid profiles of multiresistant NTS strains were heterogenous as 21 different profiles were detected in a total of 28 plasmid-bearing isolates. No direct correlation was established between antibiotic resistance patterns and plasmid profiles.

Anti-Bacterial Agents↗

PubMed1996

Biotechnological developments in Turkey.

Turkey is a country not usually associated with industrial biotechnology. However, when current research potential in universities and other R & D centers and particularly contributions made to the international literature since the mid-1980s are taken into account, high-quality international-level work is now commonplace, especially in areas such as industrial microbiology, enzyme technology, biomaterials and biological wastewater treatment. Work in plant biotechnology is at a relatively early stage, but is expected to become a rapidly developing force in the near future. The present article documents current potential in Turkey, based on significant publications produced during the last 8 years.

Animal Husbandry↗

PubMed1993

Studies on metal resistance system in Kluyveromyces marxianus.

Through preliminary plate tests, Kluyveromyces marxianus was found to be much more resistant to toxic heavy metals compared to a CUP1R strain of Saccharomyces cerevisiae. Specific growth rate and maximum dry weights affected by increasing metal concentrations were determined to obtain precise patterns of resistance. Metal biosorption was also monitored during the course of growth in synthetic media containing respective metals at 0.5 mM final concentration. Although Zn- and Co-binding was negligible, as much as 90% of silver, 60% of copper, and 65% of cadmium were found to be absorbed by the end of active growth. Analysis of the protein profiles of S. cerevisiae and K. marxianus on metal exposure suggested constitutive production of metallothionein in K. marxianus. Furthermore, a smaller protein synthesized by K. marxianus on induction by silver or cadmium accounts for the high resistance of the organism to these metals.

Drug Resistance↗

PubMed1991

Bacilysin production by Bacillus subtilis: effects of bacilysin, pH and temperature.

The dipeptide antibiotic bacilysin, when added externally to the early exponential-phase cultures, markedly limited its own synthesis. It was shown in cell-free extracts that the feedback effect does not involve the inhibition of bacilysin synthetase, the enzyme catalyzing bacilysin formation. We also studied pH and temperature dependence of bacilysin production. Production was highest at about pH 6.8 and at 25 degrees C.

Anti-Bacterial Agents↗

PubMed1991

[Plasmid analysis of antibiotic resistant Shigella isolates].

A correlation between the multiple drug resistance patterns and the plasmid profiles given by 70 clinical isolates of Shigella sonnei and Shigella flexneri was investigated in this study. The most common plasmids were purified from different isolates by electroelution and characterized via restriction endonuclease digestions.

Anti-Bacterial Agents↗

PubMed1990

Regulation of biosynthesis of bacilysin by Bacillus subtilis.

Production of the dipeptide antibiotic bacilysin by Bacillus subtilis 168 was growth associated and showed no evidence of repression by glucose or sucrose. Carbohydrates other than glucose and sucrose yielded lower specific titers of bacilysin. Bacilysin production in three such carbon sources (maltose, xylose, ribose) was delayed until growth slowed down. Ammonium salts were poor for bacilysin production when used as the sole nitrogen source. When added to the standard medium containing glutamate, they suppressed antibiotic production. Aspartate was slightly better than glutamate for antibiotic production as sole nitrogen source. No other nitrogen source tested, including inorganic, organic or complex, approached the activity of glutamate or aspartate. When added to glutamate, casamino acids, phenylalanine and alanine (a substrate of bacilysin synthetase) suppressed bacilysin production while stimulating growth. Phosphate provided for optimum growth and production at 7.5 mM and both processes were inhibited at higher concentrations. Ferric citrate stimulated growth and inhibited bacilysin production, the effects being due to both the iron and the citrate components. Elimination of ferric citrate stimulated production as did increasing the concentration of Mn to its optimum concentration of 6.6 x 10(-4) M.

Amino Acids↗