Biomedical subjects

Gülay Ozcengiz

Publications and source records attributed to Gülay Ozcengiz.

11 recordsLinked to original sources

PubMed2006

Characterization of bacteriocins from two Lactococcus lactis subsp. lactis isolates.

In this study, bacteriocins from two Lactococcus lactis subsp. lactis isolates from raw milk samples in Turkey designated OC1 and OC2, respectively, were characterized and identified. The activity spectra of the bacteriocins were determined by using different indicator bacteria including Listeria, Bacillus and Staphylococcus spp. Bacteriocins were tested for their sensitivity to different enzymes, heat treatments and pH values. Loss of bacteriocin activities after alpha-amylase treatment suggested that they form aggregates with carbohydrates. Molecular masses of the purified bacteriocins were determined by SDS-PAGE. PCR amplification was carried out with specific primers for the detection of their structural genes. As a result of these studies, the two bacteriocins were characterized as nisin and lacticin 481, respectively. Examination of plasmid contents of the isolates and the results of plasmid curing and conjugation experiments showed that in L. lactis subsp. lactis OC1 strain the 39.7-kb plasmid is responsible for nisin production, lactose fermentation and proteolytic activity, whereas the 16.0-kb plasmid is responsible for lacticin 481 production and lactose fermentation in L. lactis subsp. lactis OC2 strain.

Animals↗

PubMed2006

Enzymatic extraction of activated sludge extracellular polymers and implications on bioflocculation.

This study examines enzyme hydrolysis, a mild, effective, but a rarely used method of extracellular polymer extraction, in removing polymers from mixed culture activated sludge flocs. Two carbohydrate specific enzymes (alpha-amylase and cellulase) and a protein specific enzyme (proteinase) are used during the study. First, the kinetic aspect is investigated, then enzyme dose optimization is carried out on laboratory grown activated sludge samples cultured at solids retention times (SRT) of 4 and 20 days. A more commonly used cation exchange resin (CER) extraction technique is also employed for comparison purposes. Results indicate that the extraction of extracellular polymers by enzymes is a rather quick process reaching equilibrium within only a few hours. As the doses of enzymes are increased, the extracted polymer quantities increase up to a certain dose, beyond which not much extraction is observed. The method does not cause any significant cell lysis as measured by the viable cell counts. Carbohydrate-hydrolyzing enzymes extract small amount of proteins along with the carbohydrates and protein-hydrolyzing enzyme extracts some carbohydrates together with the proteins, indicating that proteins and carbohydrates exist bound to each other in the extracellular polymer network of sludge. Enzyme extraction generally gives a lower estimate of polymers compared to the CER method, but correctly detects the trends in the polymer quantity.

Cellulase↗

PubMed2005

Cry3Aa11: a new Cry3Aa delta-endotoxin from a local isolate of Bacillus thuringiensis.

A local isolate of Bacillus thuringiensis Mm2 had insecticidal activity against the larvae of Melolontha melolontha, Agelastica alni, Leptinotarsa decemlineata and Amphimallon solstitiale and produced a 65 kDa protein. SDS-PAGE profile of B. thuringiensis Mm2 was compared with those of 29 different Cry3Aa producers which verified Cry3Aa biosynthesis by the isolate. The cry3Aa gene of Mm2 was cloned, sequenced and the deduced amino acid sequence was compared with the cry3Aa sequences of ten different quaternary ranks. Its identity to these sequences ranged between 97.4% and 99.2%. The gene was next cloned into E. coli-Bacillus shuttle vector pNW33N and expressed at a low level in B. subtilis 168.

Amino Acid Sequence↗

PubMed2004

Cloning, characterization and heterologous expression of the aspartokinase and aspartate semialdehyde dehydrogenase genes of cephamycin C-producer Streptomyces clavuligerus.

Carbon flow through the lysine branch of the aspartate biosynthetic pathway is a rate-limiting step in the formation of cephamycin C, a broad spectrum beta-lactam antibiotic produced by Streptomyces clavuligerus. In this study, genes which encode the enzymes catalyzing the first two steps of the aspartate pathway, ask (aspartokinase) and asd (aspartate semialdehyde dehydrogenase), in S. clavuligerus NRRL 3585 were cloned and sequenced. Nucleotide sequencing and codon preference analysis revealed three complete open reading frames (ORFs). ORF2 starts within ORF1 and terminates by utilizing the same stop codon as ORF1, an arrangement typical of many ask genes. ORF3 is located 2 nucleotides downstream of ORF1,2. Database comparisons with these proteins identified ORF1 as the large (alpha) subunit of aspartokinase, ORF2 as the small (beta) subunit of aspartokinase and ORF3 as the aspartate semialdehyde dehydrogenase. The cloned genes were functionally expressed in auxotrophic Escherichia coli strains, CGSC5074 (ask(-)) and E. coli CGSC5080 (asd(-)), the two enzymes were partially purified from E. coli cell extracts and their kinetic parameters were determined. The effects of end product amino acids and diaminopimelic acid on the activity of Ask and Asd enzymes were also described.

Aspartate Kinase↗

PubMed2004

Cloning and expression of the Clostridium thermocellum L-lactate dehydrogenase gene in Escherichia coli and enzyme characterization.

The structural gene for L-lactate dehydrogenase (LDH) (EC.1.1.1.27) from Clostridium thermocellum 27405 was cloned in Escherichia coli by screening the Lambda Zap II phage library of C. thermocellum genomic DNA. In one positive clone, an open reading frame of 948 base pairs corresponded to C. thermocellum ldh gene encoding for the predicted 315-residue protein. The ldh gene was successfully expressed in E. coli FMJ39 (ldh mutant) under the lac promoter. The recombinant enzyme was partially purified from E. coli cell extracts and its kinetic properties were determined. Clostridium thermocellum LDH was shown to catalyze a highly reversible reaction and to be an allosteric enzyme that is activated by fructose-1,6-diphosphate (FDP). For pyruvate, partially purified LDH had Km and Vmax values of 7.3 mmol/L and 87 micromol/min, respectively, and in the presence of FDP, a 24-fold decrease in Km and a 5.7-fold increase in Vmax were recorded. The enzyme exhibited no marked catalytic activity for lactate in the absence of FDP, whereas Km and Vmax values were 59.5 mmol/L and 52 micromol/min, respectively, in its presence. The enzyme did not lose activity when incubated at 65 degrees C for 5 min.

Amino Acid Sequence↗

PubMed2003

The effects of insertional mutations in comQ, comP, srfA, spo0H, spo0A and abrB genes on bacilysin biosynthesis in Bacillus subtilis.

In Bacillus subtilis, two extracellular signaling peptides, ComX pheromone and CSF (competence and sporulation factor), stimulate the development of genetic competence and surfaction biosynthesis in response to high cell density (quorum sensing) by regulating the activity of transcription factor ComA. We recently showed that biosynthesis of dipeptide antibiotic bacilysin is linked to ComA and PhrC(CSF) in a Spo0K(Opp)-dependent manner by constructing phrC-, comA- and oppA-disrupted mutants of B. subtilis. In the present study, another pathway of quorum-sensing global regulation, namely, ComQ/ComX was found to be essential for bacilysin biosynthesis. ComP function in this chain was dispensable, most probably because of the existence of an alternative mean of ComA activation. The disruption of srfA operon in the bacilysin producer resulted with the bacilysin-negative phenotype; thus, our study verified that the srfA operon functions directly in the production of bacilysin. The abrB mutation suppressed the bacilysin-negative phenotype of a spo0A mutant, whereas the same mutation in the wild-type strain resulted in a significant increase in the production of bacilysin. This indicated that abrB gene product negatively controls the transcription of the gene(s) involved in bacilysin formation.

Bacillus subtilis↗

PubMed2002

Regulation of crystal protein biosynthesis by Bacillus thuringiensis: II. Effects of carbon and nitrogen sources.

The regulation of crystal protein production in Bacillus thuringiensis 81 by sources of carbon and nitrogen was investigated. The highest titers of toxin were obtained on sucrose, lactose and inulin which also supported sporulation. Whey and molasses were also potential carbon substrates for toxin production. Other carbohydrates including glucose, glycerol, maltose, starch and dextrin yielded lower amounts of toxin. Nitrogen sources were found to exert the most profound controls. Peptone was the best organic nitrogen source, supporting optimum production and sporulation as well as high cell density. The formation of CryI and CryII toxin proteins was found to be differentially regulated by the inorganic nitrogenous compounds incorporated into the medium.

Bacillus thuringiensis↗

PubMed2002

Regulation of crystal protein biosynthesis by Bacillus thuringiensis: I. Effects of mineral elements and pH.

Crystal protein synthesis by a local isolate of Bacillus thuringiensis was monitored and compared in association with growth and sporulation in media differing in mineral element content. Mg and Cu were the most important metals for the biosynthesis of 135 kDa and 65 kDa toxin components in that the former was essential and the latter was greatly stimulatory at 10(-6) to 10(-7) M concentration. Also the inclusion of Mn favored toxin production at concentrations ranging from 3 x 10(-4) to 10(-5) M. The omission of Zn and Ca had no effect on toxin formation. Crystal protein synthesis and sporulation did not generally seem to be co-regulated by the minerals as these processes responded differently to mineral levels. There was no evidence for suppression of biosynthesis by inorganic phosphate over a range of 3 to 100 mM. Crystal protein production was more efficient in buffered medium, especially when the initial pH was adjusted to 6.5.

Bacillus thuringiensis↗

PubMed2002

Characterisation of local isolates of Enterobacteriaceae from Turkey.

20 local isolates of enterics belonging to the genera Salmonella, Enterobacter, Proteus, Citrobacter from human, chicken and/or egg were characterised for their antibiotic resistance patterns, plasmid profiles, phage types, outer membrane proteins, and lipopolysaccharide patterns. Relatedness of these characteristics for epidemiological analysis was assessed. 18 (90%) strains were resistant to at least one antibiotic and those (multi-drug resistant ones) resisting to two or more antibiotics constituted 50% of all isolates. A common 54 kb plasmid was harboured by 55% of the isolates. 14 isolates showed smooth type lipopolysaccharide. 60% of the 20 isolates contained outer membrane proteins in a molecular weight range of 34.6 to 30.6 kDa. The data reveal the lack of correlation between the characteristics investigated.

Animals↗

PubMedDate not supplied

Nutritional and cultural parameters influencing antidipteran delta-endotoxin production.

In this study, various nutritional and cultural parameters influencing diptera-specific delta-endotoxin synthesis by Bacillus thuringiensis subsp. israelensis HD500 were investigated. Of various inorganic nitrogen sources, the highest yields of Cry11Aa and Cry4Ba proteins were obtained on (NH(4))(2)HPO(4). Among carbon sources, inulin, dextrin, maltose, lactose, sucrose, whey and glycerol were all stimulatory, while glucose, starch and molasses were suppressive. High concentrations of inorganic phosphate (50 to 100 mM K(2)HPO(4)) were required for an effective synthesis of Cry4Ba. Mn was the most critical element for the biosynthesis of both toxins at 10(-6) M concentration. Mg and Ca favored production when provided at 8 x 10(-3) M and 5.5 x 10(-4) M concentrations, respectively, while Fe, Zn and Cu negatively influenced biosynthesis. Cry4-toxin synthesis was best at neutral pH and also when the organism was grown at 25 degrees C. Throughout the study, the extent of growth and sporulation of the producer organism was also monitored.

Bacillus thuringiensis↗

PubMedDate not supplied

Evaluation of symbiotic effectiveness of various Rhizobium cicer strains.

Five local and seven standard strains of Rhizobium cicer were compared in terms of their efficiency in increasing the nitrogen content of the chickpea. Shoot dry weight, nodule number, nodule dry weight, protein and total nitrogen contents were taken as the parameters of plant productivity. Different combinations of the strains that were found to be promising (385, 620, Y-29 and 3379) were next employed. The maximum increase in total nitrogen content was only 3.5-fold in single infection whereas an increase as great as 35-fold was recorded for multiple infections. The double infection with Y-29 and 385 as well as the triple infection with Y-29, 620 and 3379 gave rise to the maximum values. Competitiveness of the strains in mixed infections was determined through their recovery from root surfaces and nodules and their subsequent identification. The effects of soil pH and of varying the concentration of some minerals on the outcome of symbiosis were also reported.

Evaluation Studies as Topic↗