Meltem Yesilcimen Akbas

Effectiveness of organic acid, ozonated water and chlorine dippings on microbial reduction and storage quality of fresh-cut iceberg lettuce

BACKGROUND The comparative effects of organic (citric and lactic) acids, ozone and chlorine on the microbiological population and quality parameters of fresh-cut lettuce during storage were evaluated. RESULTS Dipping of lettuce in 100 mg L(-1) chlorine solution reduced the numbers of mesophilic and psychrotrophic bacteria and Enterobacteriaceae by 1.7, 2.0 and 1.6 log(10) colony-forming units (CFU) g(-1) respectively. Treatment of lettuce with citric (5 g L(-1)) and lactic (5 mL L(-1)) acid solutions and ozonated water (4 mg L(-1)) reduced the populations of mesophilic and psychrotrophic bacteria by 1.7 and 1.5 log(10) CFU g(-1) respectively. Organic acid dippings resulted in lower mesophilic and psychrotrophic counts than ozonated water and chlorine dippings during 12 days of storage. Lactic acid dipping effectively reduced (by 2.2 log(10) CFU g(-1)) and maintained low populations of Enterobacteriaceae on lettuce for the first 6 days of storage. No significant (P > 0.05) changes were observed in the texture and moisture content of lettuce samples dipped in chlorine, organic acids and ozonated water during storage. Colour, β-carotene and vitamin C values of fresh-cut iceberg lettuce did not change significantly (P > 0.05) until day 8. CONCLUSION Lactic and citric acid and ozonated water dippings could be alternative treatments to chlorine dipping to prolong the shelf life of fresh-cut iceberg lettuce. Copyright

Application of gaseous ozone to control populations of Escherichia coli, Bacillus cereus and Bacillus cereus spores in dried figs

The effect of ozonation as a method to reduce Escherichia coli, Bacillus cereus and Bacillus cereus spores in dried figs was investigated. Dried figs were sprinkle inoculated with E. coli, B. cereus and B. cereus spores in sterile bags at a level of 10(7)microorganism g(-1), mixed and allowed to dry for 1h at 25 degrees C prior to ozonation. Inoculated samples were exposed to gaseous ozone in a chamber at 20 degrees C and 70% relative humidity. Ozone concentrations of 0.1, 0.5 and 1.0 ppm up to 360 min were used to inactivate E. coli and B. cereus while 1.0, 5.0, 7.0 and 9.0 ppm ozone concentrations for 360 min were used to treat B. cereus spores. E. coli and B. cereus counts were decreased by 3.5 log numbers at 1.0 ppm ozone concentration for 360 min ozone treatment. Up to 2 log reductions in the number of B. cereus spores were observed above 1.0 ppm ozone concentration at the end of 360 min of ozonation. No significant changes in color, pH and moisture content values of dried figs were observed after the ozonation treatments. No significant changes were found between sweetness, rancidity, flavor, appearance and overall palatability of ozonated and non-ozonated dried figs. Ozonation was found to be effective especially in reduction of vegetative cells in dried figs and a promising method for the decontamination of dried figs.

Redox-mediated interactions of VHb (Vitreoscilla haemoglobin) with OxyR: novel regulation of VHb biosynthesis under oxidative stress.

The bacterial haemoglobin from Vitreoscilla, VHb, displays several unusual properties that are unique among the globin family. When the gene encoding VHb, vgb, is expressed from its natural promoter in either Vitreoscilla or Escherichia coli, the level of VHb increases more than 50-fold under hypoxic conditions and decreases significantly during oxidative stress, suggesting similar functioning of the vgb promoter in both organisms. In the present study we show that expression of VHb in E. coli induced the antioxidant genes katG (catalase-peroxidase G) and sodA (superoxide dismutase A) and conferred significant protection from oxidative stress. In contrast, when vgb was expressed in an oxyR mutant of E. coli, VHb levels increased and the strain showed high sensitivity to oxidative stress without induction of antioxidant genes; this indicates the involvement of the oxidative stress regulator OxyR in mediating the protective effect of VHb under oxidative stress. A putative OxyR-binding site was identified within the vgb promoter and a gel-shift assay confirmed its interaction with oxidized OxyR, an interaction which was disrupted by the reduced form of the transcriptional activator Fnr (fumurate and nitrate reductase). This suggested that the redox state of OxyR and Fnr modulates their interaction with the vgb promoter. VHb associated with reduced OxyR in two-hybrid screen experiments and in vitro, converting it into an oxidized state in the presence of NADH, a condition where VHb is known to generate H2O2. These observations unveil a novel mechanism by which VHb may transmit signals to OxyR to autoregulate its own biosynthesis, simultaneously activating oxidative stress functions. The activation of OxyR via VHb, reported in the present paper for the first time, suggests the involvement of VHb in transcriptional control of many other genes as well.

Bactericidal Activity of Ozone against Escherichia coli in Whole and Ground Black Peppers

The effects of different ozone concentrations and ozonation times on the inactivation of Escherichia coli and on the organoleptic properties of whole and ground black peppers were determined. Black peppers were exposed to three different ozone concentrations (0.1, 0.5, and 1.0 ppm) for different periods (0 to 360 min) at 85% +/- 3% relative humidity (RH) and 25 +/- 0.5 degree C. An approximately 7-log CFU/g reduction of E. coli in whole black peppers was achieved by treating the samples with 0.1, 0.5, or 1.0 ppm of ozone for 360, 240, or 120 min, respectively. E. coli populations in ground black peppers were reduced from an initial count of about 7.5 log CFU/g to 1 log CFU/g after 360 min at 0.1- and 0.5-ppm ozone concentrations, whereas the same microbial reduction was obtained after 240 min at 1.0-ppm ozone concentration. An ozone concentration of 0.1 ppm for 360 min was found to be appropriate to inactivate E. coli in whole and ground black peppers without alteration of the organoleptic properties. No significant differences (P > or = 0.05) were present in the bitterness, flavor, odor, color, or overall acceptability values of the whole and ground black peppers treated with 0.1 ppm of ozone for up to 360 min.

Improved ethanol production from cheese whey, whey powder, and sugar beet molasses by “Vitreoscilla hemoglobin expressing” Escherichia coli

This work investigated the improvement of ethanol production by engineered ethanologenic Escherichia coli to express the hemoglobin from the bacterium Vitreoscilla (VHb). Ethanologenic E. coli strain FBR5 and FBR5 transformed with the VHb gene in two constructs (strains TS3 and TS4) were grown in cheese whey (CW) medium at small and large scales, at both high and low aeration, or with whey powder (WP) or sugar beet molasses hydrolysate (SBMH) media at large scale and low aeration. Culture pH, cell growth, VHb levels, and ethanol production were evaluated after 48 h. VHb expression in TS3 and TS4 enhanced their ethanol production in CW (21–419%), in WP (17–362%), or in SBMH (48–118%) media. This work extends the findings that “VHb technology” may be useful for improving the production of ethanol from waste and byproducts of various sources.

Recent trends in bioethanol production from food processing byproducts

The widespread use of corn starch and sugarcane as sources of sugar for the production of ethanol via fermentation may negatively impact the use of farmland for production of food. Thus, alternative sources of fermentable sugars, particularly from lignocellulosic sources, have been extensively investigated. Another source of fermentable sugars with substantial potential for ethanol production is the waste from the food growing and processing industry. Reviewed here is the use of waste from potato processing, molasses from processing of sugar beets into sugar, whey from cheese production, byproducts of rice and coffee bean processing, and other food processing wastes as sugar sources for fermentation to ethanol. Specific topics discussed include the organisms used for fermentation, strategies, such as co-culturing and cell immobilization, used to improve the fermentation process, and the use of genetic engineering to improve the performance of ethanol producing fermenters.

Efficient ethanol production from potato and corn processing industry waste using E. coli engineered to express Vitreoscilla haemoglobin

Engineering of ethanologenic E. coli to express the haemoglobin (VHb) from the bacterium Vitreoscilla has been shown to enhance ethanol production by fermentation of pure sugars, sugars from hydrolysis of lignocellulose, components of whey, and sugars from wastewater produced during potato processing. Here, these studies were extended to see whether the same effect could be seen when a mixture of waste materials from processing of potatoes and corn into potato and corn chips were used as sugar sources. Consistent increases in ethanol production coincident with VHb expression were seen in shake flasks at both low aeration and high aeration conditions. The ethanol increases were due almost entirely to increases in the amount of ethanol produced per unit of cell mass. The VHb strategy for increasing fermentation to ethanol (and perhaps other valuable fermentation products) may be of general use, particularly regarding conversion of otherwise discarded materials into valuable commodities.

Use of organic acids for prevention and removal of Bacillus subtilis biofilms on food contact surfaces

The efficacies of organic acid (citric, malic, and gallic acids) treatments at 1% and 2% concentrations on prevention and removal of Bacillus subtilis biofilms were investigated in this study. The analyses were conducted on microtitration plates and stainless steel coupons. The biofilm removal activities of these organic acids were compared with chlorine on both surfaces. The results showed that citric acid treatments were as powerful as chlorine treatments for prevention and removal of biofilms. The antibiofilm effects of malic acid treatments were higher than gallic acid and less than citric acid treatment. When the antibiofilm effects of these acids and chlorine on the two surfaces were compared, the prevention and removal of biofilms were measured higher on microtitration plates than those on stainless steel coupons. Higher reductions were obtained by increasing concentrations of sanitizers on 24-hour biofilm with 20-minute sanitizer treatments for removal of biofilms.

Effective ethanol production from whey powder through immobilized E. coli expressing Vitreoscilla hemoglobin.

ABSTRACT Ethanol production from whey powder was investigated by using free as well as alginate immobilized E. coli and E. coli expressing Vitreoscilla hemoglobin (VHb) in both shake flask and fermenter cultures. Media with varying levels of whey (lactose contents of 3%, 5%, 8% or 15%) and yeast extract (0.3% or 0.5%) were evaluated with fermentation times of 48–96 h. Immobilization and VHb expression resulted in higher ethanol production with all media; the increases ranged from 2% to 89% for immobilization and from 2% to 182% for VHb expression. It was determined that growth medium containing 8% lactose with 0.5% yeast extract yielded the highest ethanol production for free or immobilized strains, with or without VHb expression, in both shake flask and fermenter cultures. Immobilization with alginate was found to be a promising process for ethanol production by VHb-expressing ethanologenic E. coli.

Further investigation of the mechanism of Vitreoscilla hemoglobin (VHb) protection from oxidative stress in Escherichia coli

In Escherichia coli, Vitreoscilla hemoglobin (VHb) protects against oxidative stress, perhaps, in part, by oxidizing OxyR. Here this protection, specifically VHb-associated effects on superoxide dismutase (SOD) and catalase levels, was examined. Exponential or stationary phase cultures of SOD+ or SOD−E. coli strains with or without VHb and oxyR antisense were treated with 2 mM hydrogen peroxide without sublethal peroxide induction, and compared to untreated control cultures. The hydrogen peroxide treatment was toxic to both SOD+ and SOD− cells, but much more to SOD− cells; expression of VHb in SOD+ strains enhanced this toxicity. In contrast, the presence of VHb was generally associated in the SOD+ background with a modest increase in SOD activity that was not greatly affected by oxyR antisense or peroxide treatment. In both SOD+ and SOD− backgrounds, VHb was associated with higher catalase activity both in the presence and absence of peroxide. Contrary to its stimulatory effects in stationary phase, in exponential phase oxyR antisense generally decreased VHb levels.