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ABSTRACT: Polyphenol oxidase (PPO) catalyses oxidation of phenolics, which results in instant but differential browning in many cut fruits and vegetables, including eggplant. Eight cultivars of eggplant were characterised by their PPO specific activity, phenolic content, browning index, and PPO polymorphism. In fresh eggplant, browning was found to be dependent on both the phenolic content and PPO specific activity, whereas, total phenolic content played a major role in browning of stored fruits. Interestingly, although browning index increased in stored eggplant fruits, PPO activity reduced in four out of eight cultivars studied. Phenolic level was found to increase in all these cultivars during storage. Although a significant level of homology was observed in PPO nucleotide and conceptually translated protein sequence, two cultivars, which displayed highest PPO specific activity, differed in the 38 amino acid stretch in the peptide region 301-338.
Food Chemistry 08/2013; 139(1-4):105-14. · 3.65 Impact Factor
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ABSTRACT: Petals from different rose (Rosa centifolia) cultivars ("passion," "pink noblesse," and "sphinx") were assessed for antimutagenicity using Escherichia coli RNA polymerase B (rpoB)-based Rif (S) →Rif (R) (rifampicin sensitive to resistant) forward mutation assay against ethyl methanesulfonate (EMS)-induced mutagenesis. The aqueous extracts of rose petals from different cultivars exhibited a wide variation in their antimutagenicity. Among these, cv. "passion" was found to display maximum antimutagenicity. Upon further fractionation, the anthocyanin extract of cv. "passion" displayed significantly higher antimutagenicity than its phenolic extract. During thin-layer chromatography (TLC) analysis, the anthocyanin extract got resolved into 3 spots: yellow (Rf : 0.14), blue (Rf : 0.30), and pink (Rf : 0.49). Among these spots, the blue one displayed significantly higher antimutagenicity than the other 2. Upon high-performance liquid chromatography analysis, this blue spot further got resolved into 2 peaks (Rt : 2.7 and 3.8 min). The 2nd peak (Rt : 3.8 min) displaying high antimutagenicity was identified by ESI-IT-MS/MS analysis as peonidin 3-glucoside, whereas less antimutagenic peak 1 (Rt : 2.7) was identified as cyanidin 3, 5-diglucoside. The other TLC bands were also characterized by ESI-IT-MS/MS analysis. The least antimutagenic pink band (Rf : 0.49) was identified as malvidin 3-acetylglucoside-4-vinylcatechol, whereas non-antimutagenic yellow band (Rf : 0.14) was identified as luteolinidin anthocyanin derivative. Interestingly, the anthocyanin extracted from rose tea of cv. "passion" exhibited a similar antimutagenicity as that of the raw rose petal indicating the thermal stability of the contributing bioactive(s). The findings thus indicated the health protective property of differently colored rose cultivars and the nature of their active bioingredients.
Journal of Food Science 04/2013; · 1.66 Impact Factor
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ABSTRACT: As mutation causes many life-threatening diseases including cancer, a diet enriched with specific vegetables having potential to reduce mutagenesis possesses immense significance. In this study, 41 commonly used vegetables from diverse botanical taxa were evaluated and compared for their relative antimutagenic potential using standard assays [Escherichia coli RNA polymerase β (rpoB)-based Rif(S) → Rif(R) assay and Ames test] against known mutagens (UV, gamma radiation, 4-nitroquinoline-N-oxide and ethylmethanesulphonate). Significant differences in antimutagenicity were observed even among the cultivars within the same species, as well as at other phylogenetic levels such as genus or family. The effect of cooking in terms of boiling (aquathermal treatment), on the antimutagenicity of these vegetables, was also investigated. In majority of the cases, aquathermal treatment did not affect the antimutagenic potential. The antimutagenicity of these vegetables was not found to correlate well with their antioxidant properties.
International Journal of Food Sciences and Nutrition 01/2013; · 1.15 Impact Factor
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Asian Pacific Journal of Tropical Diseases. 12/2012;
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ABSTRACT: Eggplant (Solanum melongena) is a very rich source of polyphenol oxidase (PPO), which negatively affects its quality upon cutting and postharvest processing due to enzymatic browning. PPO inhibitors, from natural or synthetic sources, are used to tackle this problem. One isoform of PPO was 259-fold purified using standard chromatographic procedures. The PPO was found to be a 112kDa homodimer. The enzyme showed very low K(m) (0.34mM) and high catalytic efficiency (3.3×10(6)) with 4-methyl catechol. The substrate specificity was in the order: 4-methyl catechol>tert-butylcatechol>dihydrocaffeic acid>pyrocatechol. Cysteine hydrochloride, potassium metabilsulphite, ascorbic acid, erythorbic acid, resorcylic acid and kojic acid showed competitive inhibition, whereas, citric acid and sodium azide showed mixed inhibition of PPO activity. Cysteine hydrochloride was found to be an excellent inhibitor with the low inhibitor constant of 1.8μM.
Food Chemistry 10/2012; 134(4):1855-61. · 3.65 Impact Factor
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ABSTRACT: Mistranslation leads to elevated mutagenesis and replication arrest, both of which are hypothesized to result from the presence of mixed populations of wild type and mistranslated versions of DNA polymerase III subunit proteins. Consistent with this possibility, expression of missense alleles of dnaQ (which codes for the proofreading subunit ɛ) in wild type (dnaQ+) cells is shown to lead to SOS induction as well as mutagenesis. Exposure to sublethal concentrations of streptomycin, an aminoglycoside antibiotic known to promote mistranslation, also leads to SOS induction.
Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis 06/2012; 735(1-2):46-50. · 2.85 Impact Factor
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ABSTRACT: Ochratoxin A (OTA) produced in food by Aspergillus ochraceus is known to cause adverse health effects. Among the plantation products, green coffee beans are prone to fungal attack and get contaminated with OTA frequently. A fungal strain isolated from green coffee beans was characterized by morphological analyses as well as internal transcribed spacer (ITS) and 5.8S rDNA sequencing, turned out to be A. ochraceus, however, nontoxigenic. Hence, additional strains of A. ochraceus were procured and characterized for toxin production. Presterilized green coffee beans were spiked with a toxigenic strain and treated with gamma radiation. Minimum inhibitory dose (MID) of gamma radiation for 10(4) and 10(8) spores of A. ochraceus strain per 10 g of green coffee beans was found to be approximately 1 and approximately 2.5 kGy, respectively. The radiation treatment (10 kGy) almost degraded the preformed or in vitro added OTA (50 ppb) in coffee beans. OTA degradation was found to be enhanced with increase in moisture content. Cytotoxicity in terms of cell viability was found to be reduced significantly for radiation treated OTA in MTT [3-(4,5-dimethylthiazole-2yl)-2,5-diphenyl tetrazolium bromide] assay as well as flow cytometric analysis when studied using human intestinal epithelial (Int-407) cells. Similar finding was also observed with E. coli MG1655 cells. Thus the inclusion of gamma radiation treatment in the postharvest processing chain of green coffee beans could help in eliminating toxigenic fungi as well as destroying preformed OTA without affecting the sensory attributes. PRACTICAL APPLICATION: In general, mycotoxins including ochratoxin A (OTA) are highly stable to detoxifying agents. Green coffee beans are prone to fungal attack and could get frequently contaminated with the OTA due to improper drying or rehydration during storage. Gamma radiation processing of green coffee beans was found to eliminate the A. ochraceus spores as well as inactivate OTA without affecting its sensory attributes. Thus inclusion of gamma radiation in the postharvest processing chain of green coffee beans would be very useful for consumer safety and coffee trade.
Journal of Food Science 02/2012; 77(2):T44-51. · 1.66 Impact Factor
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ABSTRACT: Honey, both unifloral (Syzygiumcumini) and bifloral, demonstrated strong antimutagenicity against physical (UV, γ) and chemical (ethylmethane sulfonate) mutagens as ascertained by rpoB/RifR and Ames tests. The effect of honey was evaluated in radiation (UV or γ) exposed Escherichia coli cells for SOS response, a well known error prone repair pathway known to significantly contribute to mutagenicity by quantifying LexA repressor level, measuring cell filamentation frequency, and prophage induction by SIVET (Selectable--In-Vivo Expression Technology) assay. LexA was almost completely degraded, phenotypically long filamentous cells (∼30 μm) were formed, and SIVET induction frequency was increased in radiation exposed E. coli cultures, however, these changes were significantly inhibited in presence of honey confirming its strong antimutagenic nature. Further, rpoB/RifR mutation frequency upon UV exposure in E. coli recA- cells was found to be negligible, whereas, E. coliumuC- and umuD- knockouts showed comparatively higher mutation frequency. Honey did not show any effect on mutagenesis in these knockouts, indicating the SOS dependence of the observed mutagenesis. Honey was also found to suppress EMS induced mutagenesis but through SOS independent mechanism. Phenolics present in honey were found to be one of the important factors contributing to the antimutagenicity of honey.
Food and chemical toxicology: an international journal published for the British Industrial Biological Research Association 01/2012; 50(3-4):625-33. · 2.99 Impact Factor
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ABSTRACT: Preserving raw sugarcane juice is a challenging problem. Sugarcane juice turns brown soon after its extraction and gets spoiled due to fermentation within hours. A combination of gamma radiation (5 kGy) with permitted preservatives and low temperature storage (10 °C) could preserve raw sugarcane juice for more than a month. The preservatives used were citric acid (0.3%), sodium benzoate (0.015%), potassium sorbate (0.025%), and sucrose (10%). The treatment helped in extending the shelf life to 15 d at ambient temperature (26 ± 2 °C) and 35 d at 10 °C. The microbial load was found to be below detectable limit within this period. The biochemicals like phenolics and flavonoids were not found to be affected by addition of these preservatives. The antioxidant activities including free radical scavenging activity, nitrite scavenging activity, and reducing power were also not significantly affected. The sensory evaluation scores showed that the juice with this combination treatment was highly acceptable.
Journal of Food Science 10/2011; 76(8):M573-8. · 1.66 Impact Factor
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ABSTRACT: Xanthomonas campestris pv. glycines (Xcg), an etiological agent of the bacterial pustule disease of soybean, displayed nutritionally regulated caspase-dependent programmed cell death (PCD). Experiments showed that Xcg was under metabolic stress during PCD, as evident from the intracellular accumulation of NADH and ATP. Further, the accumulation of reactive oxygen species (ROS), as confirmed by 2',7'-dichlorofluorescein diacetate labeling, electron spin resonance spectroscopy, and scopoletin assay, was also observed along with the activation of caspase-3. ROS scavengers such as dimethylsulfoxide, glutathione, n-propyl gallate, and catalase significantly inhibited caspase biosynthesis as well as its activity, eventually leading to the inhibition of PCD. The presence of a sublethal concentration of an electron transport chain uncoupler, 2,4-dinitrophenol, was found to reduce the ROS generation and the increase in the cell survival. These results indicated that Xcg cells grown in a protein-rich medium experienced metabolic stress due to electron leakage from the electron transport chain, leading to the generation of ROS and the expression as well as the activation of caspase-3, and resulting in PCD. A bacterial DNA gyrase inhibitor, nalidixic acid, was also found to inhibit PCD. Gyrase, which regulates DNA superhelicity, and consequently DNA replication and cell multiplication, appears to be involved in the process.
FEMS Microbiology Letters 11/2010; 312(2):176-83. · 2.04 Impact Factor
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Food Chemistry. 01/2007; 101:515-523.
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ABSTRACT: Elevated mistranslation induces a mutator response termed translational stress-induced mutagenesis (TSM) that is mediated by an unidentified modification of DNA polymerase III. Here we address two questions: (i) does TSM result from direct polymerase corruption, or from an indirect pathway triggered by increased protein turnover? (ii) Why are homologous recombination functions required for the expression of TSM under certain conditions, but not others? We show that replication of bacteriophage T4 in cells expressing the mutA allele of the glyVtRNA gene (Asp-Gly mistranslation), leads to both increased mutagenesis, and to an altered mutational specificity, results that strongly support mistranslational corruption of DNA polymerase. We also show that expression of mutA, which confers a recA-dependent mutator phenotype, leads to increased lambdoid prophage induction (selectable in vivo expression technology assay), suggesting that replication fork collapse occurs more frequently in mutA cells relative to control cells. No such increase in prophage induction is seen in cells expressing alaVGlu tRNA (Glu-->Ala mistranslation), in which the mutator phenotype is recA-independent. We propose that replication fork collapse accompanies episodic hypermutagenic replication cycles in mutA cells, requiring homologous recombination functions for fork recovery, and therefore, for mutation recovery. These findings highlight hitherto under-appreciated links among translation, replication and recombination, and suggest that translational fidelity, which is affected by genetic and environmental signals, is a key modulator of replication fidelity.
Molecular Microbiology 12/2006; 62(6):1752-63. · 5.01 Impact Factor
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ABSTRACT: In earlier studies from this laboratory, Xanthomonas campestris pv. glycines was found to exhibit a nutrition stress-related postexponential rapid cell death (RCD). The RCD was exhibited in protein-rich media but not in starch or other minimal media. This RCD in X. campestris pv. glycines was found to display features similar to those of the programmed cell death (PCD) of eukaryotes. Results of the present study showed that the observed RCD in this organism is both positively and negatively regulated by small molecules. The amino acids glycine and l-alanine as well as the D isomers of valine, methionine, and threonine were found to induce the synthesis of an active caspase-3-like protein that was associated with the onset of RCD. Addition of pyruvate and citrate to the culture medium induced both the synthesis of active caspase-3-like protein and RCD. Higher levels of intracellular accumulation of pyruvate and citrate were also observed under conditions favoring RCD. On the other hand, dextrin and maltose, the hydrolytic products of starch, inhibited the synthesis of the caspase-3-like protein. Addition of glucose and cyclic AMP (cAMP) to the RCD-favoring medium prevented RCD. Glucose, cAMP, caffeine (a known inhibitor of a phosphodiesterase that breaks down cAMP), and forskolin (from the herb Coleus forskholii, known to activate the enzyme adenylate cyclase that forms cAMP) inhibited the caspase enzyme activity in vivo and consequently the RCD process. The addition of glucose and other inhibitors of RCD enhanced intracellular cAMP accumulation. This is the first report demonstrating the involvement of small molecules in the regulation of nutrition stress-related stationary-phase rapid cell death in X. campestris pv. glycines, which is programmed.
Journal of Bacteriology 09/2006; 188(15):5408-16. · 3.83 Impact Factor
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ABSTRACT: Xanthomonas campestris pv. glycines strain AM2 (XcgAM2), the etiological agent of bacterial pustule disease of soybean, exhibited post-exponential rapid cell death (RCD) in LB medium. X. campestris pv. malvacearum NCIM 2310 and X. campestris NCIM 2961 also displayed RCD, though less pronouncedly than XcgAM2. RCD was not observed in Pseudomonas syringae pv. glycines, or Escherichia coli DH5alpha. Incubation of the post-exponential LB-grown XcgAM2 cultures at 4 degrees C arrested the RCD. RCD was also inhibited by the addition of starch during the exponential phase of LB-growing XcgAM2. Protease negative mutants of XcgAM2 were found to be devoid of RCD behavior observed in the wild type XcgAM2. While undergoing RCD, the organism was found to transform to spherical membrane bodies. The presence of membrane bodies was confirmed by using a membrane specific fluorescent label, 1,6-diphenyl 1,3,5-hexatriene (DPH), and also by visualizing these structures under microscope. The membrane bodies of XcgAM2 were found to contain DNA, which was devoid of the indigenous plasmids of the organism. The membrane bodies were found to bind annexin V indicative of the externalization of membrane phosphatidyl serine. Nicking of DNA in XcgAM2 cultures undergoing RCD in LB medium was also detected using a TUNEL assay. The RCD in XcgAM2 appeared to have features similar to the programmed cell death in eukaryotes.
The Journal of General and Applied Microbiology 05/2002; 48(2):67-76. · 0.98 Impact Factor
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ABSTRACT: Xanthomonas campestris pv. glycines strain AM2 (XcgAM2), the aetiological agent of bacterial pustule disease of soybean, as well as some other strains of Xanthomonas including X. campestris pv. malvacearum NCIM 2310 and X. campestris NCIM 2961, exhibited post-exponential rapid cell death (RCD) in Luria-Bertani (LB) medium. RCD was not displayed by Xanthomonas strains while growing in starch medium. Addition of starch to LB culture of XcgAM2 at any point of incubation during the exponential growth was found to arrest the onset of RCD. RCD in this organism was found to be associated with the synthesis of an endogenous enzyme similar to human caspase-3, a known marker of apoptosis in eukaryotes. On sodium dodecyl sulphate polyacrylamide gel elecrophoresis (SDS-PAGE) the XcgAM2 caspase appeared to run along a 55 kDa protein molecular weight marker. The caspase-3-like protein was detected in all Xanthomonas strains tested. RCD was not detected in Escherichia coli cultures in LB medium. The caspase-3-like enzyme activity or pro-tein was also found to be absent in this bacterium. Caspase-3-like protein or Xanthomonas caspase was detected only in the cells of XcgAM2 growing in LB medium and not in those growing in starch medium. The Xanthomonas caspase protein appeared in cells at around 4 h of incubation, and peaked at around 24 h, before finally disappearing at around 54 h of incubation. However, caspase enzyme activity was detected only 12-13 h after incubation and peaked around 18-20 h. Addition of starch at the beginning or during the period of exponential growth in LB cultures of XcgAM2 terminated the synthesis of this protein. It is presumed that starch acted as the repressor of biosynthesis of the Xanthomonas caspase, thereby preventing the organism from undergoing RCD. The cells undergoing RCD also displayed the other markers of eukaryotic apoptosis. These included binding of annexin V to plasma membrane of cells undergoing RCD and the presence of nicked DNA in culture supernatant as evidenced by the TUNEL (terminal deoxynucleotidyl transferase dUTP nick-end labelling) assay. Caspase-negative mutants of XcgAM2 did not display post-exponential RCD. The importance of RCD in Xanthomonas life cycle is not yet clear, however the phenomenon appears to have similarities with eukaryotic apoptosis.
Molecular Microbiology 05/2002; 44(2):393-401. · 5.01 Impact Factor
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ABSTRACT: A microbiological method for the detection of irradiated spices is proposed. The method involves harvesting of residual microflora from the spice sample and allowing it to grow in a nutrient medium. After a fixed interval of incubation, turbidity developed is measured as absorbency at 600 nm (A600). The differences between the A600 values obtained from irradiated and nonirradiated samples are large enough to allow identification of the former. The growth profiles of the harvested microflora from irradiated spices were found to be different. A dose-dependent increase in lag phase was observed with the harvested residual microflora from irradiated samples. The proposed method is much simpler and less cumbersome compared to the DEFT/APC method recommended currently for detection of irradiated spices. Keywords: Irradiated spices; detection methods; microbiological; turbidimetry
11/1998;
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ABSTRACT: The exposure of plasmid pUC18 and pBR322 DNA to high hydrostatic pressure increased the ability of plasmids to transform competent Escherichia coli cells. For pUC18 plasmid, a pressure of 400 MPa, and for pBR322, a pressure of 200 MPa was found to provide the highest transformation efficiency. The DNA duplexes of the two plasmids were found to be the most stable for melting conditions at these pressures. At pressures higher than these, both the stability of the duplex DNA and the transformation efficiency were affected. The stabilizing effect of high hydrostatic pressure on the hydrogen bond may be responsible for the observed increase in transformation efficiency of the pressure-exposed plasmid DNA. The possibility of pressure-induced changes in the structure and conformation of DNA was studied using various techniques. In agarose gel electrophoresis, pressure-treated plasmids (pUC18 at 400 MPa and pBR322 at 200 MPa) consistently showed visibly distinct higher mobility compared to untreated plasmids. Pressure-treated pUC18 as well as pBR322 DNA showed significant reduction in ethidium bromide binding as is evident from the reduced intensity of fluorescence of the dye bound pressure-treated DNA. Spectroscopic studies using circular dichroism and Fourier transform infrared (FTIR) spectroscopy also showed significant differences in the absorption profiles of pressure-treated plasmids as compared to an untreated control. These studies revealed that the pressure-induced changes in the conformation of these DNAs may be responsible for the observed increase in the transformation ability of the plasmids. On the other hand, the exposure of competent cells of E. coli to a high hydrostatic pressure of 50 MPa not only reduced their colony-forming ability but also drastically reduced their ability to take up plasmid DNA.
The Journal of General and Applied Microbiology 09/1997; 43(4):199-208. · 0.98 Impact Factor
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ABSTRACT: Gamma radiation was found to be an effective tool for hygienization of municipal wastewater sludge. The sludge received from the primary settling tank of a municipal wastewater treatment plant was gamma irradiated using a cobalt-60 source in a sludge hygienization research irradiator. The process parameters were adjusted to effectively eliminate coliform bacteria in the sludge and to prevent their regrowth. Irradiated sludge was found to be free of fecal coliform and could be directly disposed after drying in a landfill or used as manure. It could also be used as a medium for growth of Rhizobium sp for obtaining a bio-fertilizer.
Water Environment Research 77(5):472-9. · 0.88 Impact Factor
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ABSTRACT: Gamma radiation is known to inactivate microorganisms in various foods and thus ensures their microbial safety. In the present study, process parameters were standardized for achieving microbial decontamination of honey of Indian origin. Study was also carried out to examine the effect of gamma radiation treatment on the biochemical, antioxidant, antibacterial, and organoleptic attributes of the honey. A 15 kGy dose of gamma radiation was found to be sufficient for complete microbial decontamination of honey including spores, thus improving its microbial safety without affecting the quality attributes.
Journal of Food Science 75(1):M19-27. · 1.66 Impact Factor
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ABSTRACT: A comprehensive study was carried out to assess the microbiological and biochemical characteristics of four herbals, namely, rose (Rosa centifolia), guggul (Commiphora mukul), chirata (Swertia chirayita), gulvel (Tinospora cordifolia) and four herbal formulations rasayan, shatpatryadi, scrub and kashayam. Total aerobic plate count (TAPC) was in the range of 3–7 log cfu/g, whereas, presumptive coliform count in many of these samples was in the range 2–6 log cfu/g. The IMViC (indole, methyl red, Voges-Proskauer, citrate) analysis and molecular characterisation (16S rDNA sequencing) ascertained the presence of Escherichiacoli in some of the samples. A gamma radiation dose of up to 10 kGy was found to be sufficient for complete microbial decontamination without affecting the bioactive properties of herbal formulations, including antioxidant potential, which was high in rasayan, shatpatryadi, scrub, rose, and guggul. The antioxidant property of these herbals could be attributed to components such as phenolics, flavonoids and colour pigments.
Food Chemistry. 119(1):328-335.
