Microbiological contamination of fresh-cut produce in Korea.

This study evaluated the microbiological contamination of fresh-cut produce in Korea. A total of 108 fresh-cut vegetables and fruits were surveyed for the aerobic mesophilic (AM) count, aerobic psychrophilic (AP) count, total coliform, generic Escherichia coli, yeast and mold, and foodborne pathogens. AM counts ranged from 1.00 to 7.36 log CFU/g, and AP counts showed very similar results as AM counts. For total coliform and generic E. coli, 53.7% and 9.3% of the samples were detected, respectively. For foodborne pathogens, none of the samples were identified as E. coli O157:H7 or Salmonella spp. However, S. aureus and B. cereus was detected in 5.6% and 6.5% of the samples, respectively. Although the contamination level has varied widely, samples with high bacterial counts, such as julienned green onion, bell pepper, carrot, and mixed sprout, should be implemented with strict control measures.

Microbial inactivation in fresh and minimally processed foods by intense pulsed light (IPL) treatment.

The purposes of this study were to evaluate the inactivation effects of intense pulsed light (IPL) on indigenous and inoculated microorganisms in fresh and minimally processed foods and the industrial applicability of this nonthermal sterilization method. The samples were treated with IPL by varying the treatment time and voltage. The inactivation effect tended to increase as the treatment conditions increased. Further, indigenous microorganisms showed a lower inactivation level than inoculated microorganisms, E. coli ATCC 25922, due to the variability of indigenous microorganisms and their properties. Chopped garlic showed a higher E. coli inactivation effect (2.65 log reduction after 0.185 J/cm2 of IPL) than peeled garlic (1.21 log reduction) due to its larger surface area. The manila clam showed a lower E. coli inactivation (0.93 log reduction) effect than squid (1.84 log reduction) due to its rougher surface. After the IPL treatment, there was no significant difference in temperature, moisture content, and color.

Bactericidal effect of intense pulsed light on seeds without loss of viability.

This study investigated the microbial inactivation effects of intense pulsed light (IPL) treatment as an alternative to chemical treatment for decontaminating the radish and pak choi seeds. The f R values (which indicate the resistance to IPL treatment) for radish and pak choi seeds were 24.50, 20.81 J/cm2, respectively. This resistance exhibited by seeds to IPL treatment is related to their surface roughness. Their Rq (the root-mean-square roughness), average surface roughness (Ra), and 10-point height roughness (Rz) values indicate that each crevice on a rough surface could shelter microorganisms from IPL. Viability tests of seeds exposed to IPL treatment indicated that the average germination rates of treated seeds exceeded 85% on day 3 of germination, which is considered as an acceptable criterion for germination. Also, on day 5 of germination the average shoot lengths of sprouts exposed to IPL did not differ significantly from those of untreated seeds.

Sporicidal Activities of Various Surfactant Components against Bacillus subtilis Spores.

The sporicidal activities against Bacillus subtilis spores of surfactant components with hydrophilic and hydrophobic properties that can lead to the denaturation of various proteins comprising the spore structure were investigated. The reduction in spore numbers by each of the surfactant components bornyl acetate, geranyl acetate, pinene, p-cymene, camphene, citral, 2,3-dihydrobenzofuran, polylysine, and thiamine dilaurylsulfate at 1% was estimated at 1 to 2 log CFU/ml. The average hydrophilelipophile balance value of surfactants with sporicidal activity causing a reduction of 1 to 2 log CFU/ml was 9.3, with a range from 6.7 to 15.8, which is similar to the values of various chemical surfactants of 9.6 to 16.7. The results also showed that the surfactants that were hydrophobic were more effective than those that were hydrophilic in killing B. subtilis spores. Furthermore, the sporicidal effect of surfactants like geranyl acetate and γ-terpinene was significantly enhanced in the presence of a germinant, because L-alanine and synergistic cofactors (e.g., K(+) ions) trigger cortex hydrolysis in spores.

Extraction characteristics of subcritical water depending on the number of hydroxyl group in flavonols.

This study compared the efficiencies of using subcritical water, hot water, and organic solvents to extract flavonols from black tea, celery, and ginseng leaf. The effect of key operating conditions was determined by varying the temperature (110-200°C), extraction time (5-15min), and pressure (about 10MPa) and the extracts were analysed quantitatively using HPLC. The yields of myricetin, quercetin, and kaempferol from plants were maximal at extraction temperatures of 170°C, 170°C and 200°C, respectively, and they depend on the number of hydroxyl groups included in the chemical structure of the flavonols, with more of those with fewer hydroxyl (OH) groups attached being extracted at higher temperatures. The results also showed that the yields of flavonols by subcritical water extraction were 2.0- to 22.7- and 1.8- to 23.6-fold higher than those obtained using the ethanol and methanol as traditional extraction methods, respectively.

Optimization of subcritical water extraction of flavanols from green tea leaves.

The subcritical-water extraction (SWE) of six kinds of flavanols from green tea leaves and the effect of extraction conditions were investigated by varying the temperature and time. The maximum yield of total flavanols, 71.36 ± 4.23 mg/g green tea leaves (mean ± SD), was obtained under extraction temperature/time conditions of 150 °C/5 min. The efficiency of SWE for total flavanols was slightly higher than that of the conventional extraction solvents such as methanol and ethanol. The extraction of flavanols via SWE was specifically adequate for epimer structures such as catechin, catechin gallate, and gallocatechin gallate due to the epimerization of epicatechins. The extraction efficiency of epimers was increased at temperatures up to 170 °C, whereas that of epicatechins was decreased. Thus, most epicatechins were converted to epimers during SWE, leading to some flavanol destruction at high temperatures, except when a short extraction time of 5 min was used.

Relationship analysis between flavonoids structure and subcritical water extraction (SWE).

Subcritical water (about 10MPa) is an excellent solvent for extracting non-polar flavonoids by varying the temperature-dependent dielectric constant. This study determined the optimum conditions for subcritical water extraction (SWE), such as the time and temperature, for extracting flavonoids from eight plants, and their dependence on the chemical structure of flavonoids (polarity of side chains and the presence of sugar, and double bonds). Flavonoids having an OH side chain (quercetin at 170°C/10min) were optimally extracted at lower temperatures than O-CH3 (isorhamnetin at 190°C/15min) and H (kaempferol at 190°C/15min) side chains. The optimal temperatures of the glycoside forms including sugar, such as quercitrin (110°C/5min), spiraeoside (150°C/15min), and isoquercitrin (150°C/15min), were lower than of the less-polar aglycones (170°C/10min and 190°C/15min). Apigenin, having double bonds, was extracted well at a higher temperature (190°C/15min) than naringenin (170°C/15min) in SWE.

Optimization of processing conditions for the sterilization of retorted short-rib patties using the response surface methodology.

The aim of this study was to determine the optimum sterilization conditions for short-rib patties in retort trays by considering microbiological safety, nutritive value, sensory characteristics, and textural properties. In total, 27 sterilization conditions with various temperatures, times, and processing methods were tested using a 3(3) factorial design. The response surface methodology (RSM) and contour analysis were applied to find the optimum sterilization conditions for the patties. Quality attributes were significantly affected by the sterilization temperature, time, and processing method. From RSM and contour analysis, the final optimum sterilization condition of the patties that simultaneously satisfied all specifications was determined to be 119.4°C for 18.55min using a water-cascading rotary mode. The findings of the present study suggest that using optimized sterilization conditions will improve the microbial safety, sensory attributes, and nutritional retention for retorted short-rib patties.

An adjunctive therapeutic vaccine against reactivation and post-treatment relapse tuberculosis.

Preventing latently infected or inadequately treated individuals from progressing to active disease could make a major impact on tuberculosis (TB) control worldwide. The purpose of this study was to evaluate a new approach to prevent reactivation and TB relapse that combines drug treatment and vaccination. Mycobacterium tuberculosis harbors a gene called mce1R that, in vivo, negatively regulates a 13-gene cluster called the mce1 operon. In a Cornell mouse model, BALB/c mice infected with M. tuberculosis H37Rv disrupted in mce1R consistently develop latent infection and reactivation disease. We used this new mouse model to test a recombinant M. tuberculosis cell wall protein (Mce1A), encoded by a gene in the mce1 operon, for its ability to prevent post-treatment TB. At 32 weeks of follow-up, a complete sterilizing protection was observed in lungs of the vaccinated mice. Mce1A but not phosphate-buffered saline administered intraperitoneally during the period of latent infection prevented disease progression and proliferation of M. tuberculosis mce1R mutant. The only visible lung lesions in vaccinated mice included small clusters of lymphocytes, while the unvaccinated mice showed progressively enlarging granulomas comprised of foamy macrophages surrounded by lymphocytes. The combination of anti-TB drugs and a vaccine may serve as a powerful treatment modality against TB reactivation and relapse.

Preparation and properties of polypropylene/clay nanocomposites for food packaging.

Polypropylene (PP)/clay nanocomposites based on PP, organic clay (montmorillonite; MMT), and maleated polypropylene (MAPP) were prepared by melt compounding. The mechanical, thermal, morphological, and gas barrier properties of the resulting PP/clay nanocomposites were investigated at varying concentrations of the components for food packaging. The results revealed that the mechanical strengths, including tensile, flexural, and Izod impact strength, were increased for PP/clay nanocomposites compared to neat PP. The thermal properties showed a tendency for the melting and degradation temperatures to increase with the clay concentration. The effect of the compatibilizer was observed using scanning electron microscopy (SEM). The X-ray diffraction pattern of the nanocomposites revealed increased d-spacing of the MMT layers, indicating that the compatibility of neat PP and clay was improved by the addition of MAPP, and the intercalation and partial exfoliation of the layers. The use of clay increased the mobility distance of the gas molecules, leading to the oxygen permeability of neat PP being reduced by 26% to 55%.

Posttreatment reactivation of tuberculosis in mice caused by Mycobacterium tuberculosis disrupted in mce1R.

BACKGROUND: The reactivation of tuberculosis arises in persons who are latently infected and in those who have been previously treated. The mechanism of the reactivation of tuberculosis in either situation is not well understood. A 13-gene mce1 operon of Mycobacterium tuberculosis was previously shown to be associated with latent infection in mice and may also play a role in reactivation. METHODS: We tested mce1 operon M. tuberculosis mutants in a Cornell mouse model to examine disease progression and reactivation. RESULTS: In BALB/c mice, the wild-type, mce1 operon mutant, and mce1R (negative transcriptional regulator of the mce1 operon) mutant M. tuberculosis strains were equally susceptible to orally administered isoniazid and pyrazinamide. However, after cessation of the treatment, the mce1R mutant rapidly and progressively proliferated in mouse lungs and spleens, whereas the other strains remained latent. The reactivation of the mce1R mutant was associated with disease progression in the mouse lungs. CONCLUSION: This observation demonstrates that the constitutive expression of the mce1 genes by M. tuberculosis in the latent state can cause a reactivation of tuberculosis. The constitutive expression of the mce1 genes in the mce1R mutant may allow this mutant to maintain its lipid metabolism, enabling it to survive long-term and proliferate inside granulomas.

Enhanced production of gamma-aminobutyric acid using rice bran extracts by Lactobacillus sakei B2-16.

An efficient and simple fermentation process was developed for the production of gamma-amminobutyric acid (GABA) by Lactobacillus sakei B2-16. When the L. sakei B2-16 was cultivated in the rice bran extracts medium containing 4% sucrose, 1% yeast extract and 12% monosodium glutamate, the maximum GABA concentration reached 660.0 mM with 100% conversion yield, showing the 2.4-fold higher GABA concentration compared to the modified MRS medium without the rice bran extracts. The GABA production was scaled-up from a laboratory scale (5 L) to a pilot (300 L) and a plant scales (5,000 L) to investigate the application possibility of GABA production to industrial fields. The GABA production at the pilot and plant scales was similar to the laboratory scale using rice bran extracts medium which could be effective for the low-cost production of GABA.

Production of D-tagatose at high temperatures using immobilized Escherichia coli cells expressing L-arabinose isomerase from Thermotoga neapolitana.

Escherichia coli cells expressing L-arabinose isomerase from Thermotoga neapolitana (TNAI) were immobilized in calcium alginate beads. The resulting cell reactor (2.4 U, t (1/2) = 43 days at 70 degrees C) in a continuous recycling mode at 70 degrees C produced 49 and 38 g D-tagatose/l from 180 and 90 g D-galactose/l, respectively, within 12 h.

Purification and characterization of a fibrinolytic subtilisin-like protease of Bacillus subtilis TP-6 from an Indonesian fermented soybean, Tempeh.

We have isolated a bacterium (TP-6) from the Indonesian fermented soybean, Tempeh, which produces a strong fibrinolytic protease and was identified as Bacillus subtilis. The protease (TPase) was purified to homogeneity by ammonium sulfate fractionation and octyl sepharose and SP sepharose chromatography. The N-terminal amino acid sequence of the 27.5 kDa enzyme was determined, and the encoding gene was cloned and sequenced. The result demonstrates that TPase is a serine protease of the subtilisin family consisting of 275 amino acid residues in its mature form. Its apparent K (m) and V (max) for the synthetic substrate N-succinyl-Ala-Ala-Pro-Phe-pNA were 259 microM and 145 micromol mg(-1) min(-1), respectively. The fibrinogen degradation pattern generated by TPase as a function of time was similar to that obtained with plasmin. In addition, N-terminal amino acid sequence analysis of the fibrinogen degradation products demonstrated that TPase cleaves Glu (or Asp) near hydrophobic acids as a P1 site in the alpha- and beta-chains of fibrinogen to generate fragments D', E', and D' similar to those generated by plasmin. On plasminogen-rich fibrin plates, TPase did not seem to activate fibrin clot lysis. Moreover, the enzyme converted the active plasminogen activator inhibitor-1 to the latent form.

Nisin biosynthesis and its properties.

The antimicrobial peptide, nisin, produced by several strains of Lactococcus lactis, which belongs to the Class I bacteriocins called lantibiotics, is a small (3.4 kDa), 34-amino acid, cationic, hydrophobic peptide and has the five characteristic (beta-methyl)lanthionine rings formed by significant post-translational modification. A cluster of 11 genes has been involved in the biosynthesis of nisin and are proposed to be transcriptionally arranged as nisA(Z)BTCIP, nisRK, and nisFEG. The biosynthesis of nisin is regulated in a growth-phase-dependent manner including nisin-mediated induction which occurs via NisRK two-component regulatory system. This review outlines some of the more recent developments in the properties, regulation and applications of nisin biosynthesis.

Enhanced nisin production by increasing genes involved in nisin Z biosynthesis in Lactococcus lactis subsp. lactis A164.

Nisin Z production in Lactococcus lactis subsp. lactis A164 was improved by introducing multicopy genes, nisZ, nisRK, or nisFEG, involved in nisin biosynthesis into A164 strain. A similar growth profile was obtained from all strains tested. However, the cells expressing nisRK produced 25,000 AU nisin Z ml(-1) compared to 16,000 AU ml(-1) by the control strain. Northern blot analysis revealed that over-expression of nisRK promoted the transcription of the nisZ gene. The A164 strain expressing multicopy nisFEG also had an increased nisin Z production (25,000 AU ml (-1)) but produced the nisin more slowly than the cells expressing multicopy nisRK.

Simple one-step purification of nisin Z from unclarified culture broth of Lactococcus lactis subsp. lactis A164 using expanded bed ion exchange chromatography.

A simple one-step purification method, using expanded bed, ion-exchange chromatography, for the fractionation of nisin Z produced by Lactococcus lactis subsp. lactis A164 was developed. The highest dynamic binding capacity (0.92) of the adsorbent was obtained at a superficial velocity of 367 cm h(-1), resulting in approx. 2.7-fold bed expansion. The range of pH for the maximum adsorption was 3-4. The isocratic elution with 0.15 M NaCl led to approx. >90% recovery. Single-step purification of nisin Z from unclarified A164 culture broth resulted in 31-fold purification with a 90% yield.

Antagonism of Helicobacter pylori by bacteriocins of lactic acid bacteria.

Antimicrobial activity of seven bacteriocins produced by lactic acid bacteria against Helicobacter pylori strains (ATCC 43504, Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH [DSM] 4867, DSM 9691, and DSM 10242) was investigated in vitro using a broth microdilution assay. The bacteriocins chosen for the study were nisin A; lacticins A164, BH5, JW3, and NK24; pediocin PO2; and leucocin K. Antimicrobial activity of the bacteriocins varied among the H. pylori strains tested, of which strain ATCC 43504 was the most tolerant. Among the bacteriocins tested, lacticins A164 and BH5 produced by Lactococcus lactis subsp. lactis A164 and L. lactis BH5, respectively, showed the strongest antibacterial activity against H. pylori strains. MICs of the lacticins against H. pylori strains, when assessed by the critical dilution micromethod, ranged from 0.097 to 0.390 mg/liter (DSM strains) or from 12.5 to 25 mg/liter (ATCC 43504), supporting the strain-dependent sensitivity of the pathogen. Pediocin PO2 was less active than the lacticins against four strains of H. pylori, and leucocin K was the least active peptide, with no inhibition toward H. pylori ATCC 43504. Anti-Helicobacter activity of lacticin A164 was dependent on initial inoculum size as well as concentration of the bacteriocin added.

Influence of growth conditions on the production of a nisin-like bacteriocin by Lactococcus lactis subsp. lactis A164 isolated from kimchi.

The influence of growth parameters on the fermentative production of a nisin-like bacteriocin by Lactococcus lactis subsp. lactis A164 isolated from kimchi was studied. The bacteriocin production was greatly affected by carbon and nitrogen sources. Strain A164 produced at least 4-fold greater bacteriocin in M17 broth supplemented with lactose than other carbon sources. The amount of 3% yeast extract was found to be the optimal organic nitrogen source. While the maximum biomass was obtained at 37 degrees C, the optimal temperature for the bacteriocin production was 30 degrees C. The bacteriocin production was also affected by pH of the culture broth. The optimal pH for growth and bacteriocin production was 6.0. Although the cell growth at pH 6.0 was nearly the same level at pH 5.5 and 6.5, the greater bacteriocin activity was observed at pH 6.0. Exponential growth took place only during an initial period of the cultivation, and then linear growth was observed. Linear growth rates increased from 0.160 g(DCW) x l(-1) x h(-1) to 0.245 g(DCW) x l(-1) x h(-1) with increases in lactose concentrations from 0.5 to 3.0%. Maximum biomass was also increased from 1.88 g(DCW) x l(-1) to 4.29 g(DCW) x l(-1). However, increase in lactose concentration did not prolong the active growth phase. After 20 h cultivation, cell growth stopped regardless of lactose concentration. Production of the bacteriocin showed primary metabolic kinetics. However, bacteriocin yield based on cell mass increased greatly during the late growth phase. A maximum activity of 131x10(3) AU x ml(-1) was obtained at early stationary growth phase (20 h) during the batch fermentation in M17L broth (3.0% lactose) at 30 degrees C and pH 6.0.

Weissella kimchii sp. nov., a novel lactic acid bacterium from kimchi.

A gram-positive, catalase-negative, non-sporulating, facultatively anaerobic, short rod-shaped bacterium, with cells measuring 0.3-0.5 x 1-2 microm and designated strain CHJ3T, was isolated from partially fermented kimchi, a traditional Korean fermented vegetable food. The strain produced CO2 gas, D-lactate from glucose and dextran from sucrose and hydrolysed aesculin and arginine. It also fermented N-acetylglucosamine, amygdalin, arbutin, cellobiose, D-fructose, galactose, beta-gentiobiose, gluconate, D-glucose, maltose, D-mannose, salicin, sucrose and D-xylose. The G+C content of the DNA was 48.2 mol%. Phylogenetic analysis of 16S rRNA showed that strain CHJ3T is a member of the genus Weissella. The nearest phylogenetic relative of strain CHJ3T was Weissella confusa, with 16S rRNA similarity of 98.3%. However, strain CHJ3T could be differentiated from W. confusa on the basis of some phenotypic characteristics, analysis of whole-cell protein patterns and DNA-DNA hybridization data. These data suggest that strain CHJ3T be classified in the genus Weissella as a novel species, Weissella kimchii sp. nov. The type strain is CHJ3T (= KCCM 41287T = DSM 14295T = KCTC 3746T).