Cells of Acinetobacter sp. strain JC1 DSM 3803, an aerobic monoxide-oxidizing bacterium, growing on glucose exhibited high catalase activity at the mid-exponential growth phase. The enzyme activity decreased gradually after then until the early stationary phase, increased again at the mid-stationary phase, and then decreased again thereafter. Cells growing on glucose was found to contain three kinds of catalses. Cat1, Cat2 and Cat3. The activities of Cat1 and Cat3 did change significantly during growth, but that of Cat2 exhibited significant variation. Cat3 was found to present only in cells growing on glucose, but not in cells growing on carbon monoxide of methanol. The activities of call and Cat3 in cell-free extracts were stable upon treatment with ethanol and chloroform, but decreased to some extent when the enzymewere treated with 2mM $H_2O_2$ and/or 3-amino-1,2,4-triazole (AT). Cat2 was found to be extremely sensitive to the ethanol-chloroform and $H_2O_2$ treatments, but was insensitive to the AT treatment. Cat1 exhibited enzyme activity after incubation for 1 min at 80$^{\circ}C$. Cat2 and Cat3 did not show enzyme activity after incubation for 1 min at 60$^{\circ}C$ and 70$^{\circ}C$, respectively. Cat2 was found to have peroxidase activity. Cat3 was purified to homogenity in seven steps. The molecular weight of the native enzyme was estimated to be 150,000. Sodium dodecyl sulfate-gel electrophoresis revealed two identical subunits of molecular weight 65,000. The enzyme was found to show two $K_m$ values of 39 mM and 58mM. The optimal pH for the enzyme activity was 7.0, but the activities at pH 6.0, 8.0, and 9.0, were found to be comparable to that at the optimal pH. The optimal temperature for the enzyme activity was found to be 40$^{\circ}C$. The enzyme also exhibited strong activity at 20$^{\circ}C$, 30$^{\circ}C$, and 50$^{\circ}C$. The purified enzyme was not affected by the ethanol-chloroform treatment. The enzyme, howerver, showed less than 10% of the original activity when it was treated with 12 mN AT, 0.1 mM $NaN_3$ of 1mM KCN.
A constructed sea stream in Yeongdo, Busan, Republic of Korea is mostly static due to the lifted stream bed and tidal characters, and receives domestic wastewater nearby, causing a consistent odor production and water quality degradation. Bioaugmentation of a microbial consortium was proposed as an effective and economical restoration technology to restore the polluted stream. The microbial consortium activated on site was augmented on a periodic basis (7~10 days) into the most polluted site (Site 2) which was chosen considering the pollution level and tidal movement. Physicochemical parameters of water qualities were monitored including pH, temperature, DO, ORP, SS, COD, T-N, and T-P. COD and microbial community analyses of the sediments were also performed. A significant reduction in SS, COD, T-N, and COD (sediment) at Site 2 occurred showing their removal rates 51%, 58% and 27% and 35%, respectively, in 13 months while T-P increased by 47%. In most of the test sites, population densities of sulfate reducing bacterial (SRB) groups (Desulfobacteraceae_uc_s, Desulfobacterales_uc_s, Desulfuromonadaceae_uc_s, Desulfuromonas_g1_uc, and Desulfobacter postgatei) and Anaerolinaeles was observed to generally decrease after the bioaugmentation while those of Gamma-proteobacteria (NOR5-6B_s and NOR5-6A_s), Bacteroidales_uc_s, and Flavobacteriales_uc_s appeared to generally increase. Aerobic microbial communities (Flavobacteriaceae_uc_s) were dominant in St. 4 that showed the highest level of DO and least level of COD. These microbial communities could be used as an indicator organism to monitor the restoration process. The alpha diversity indices (OTUs, Chao1, and Shannon) of microbial communities generally decreased after the augmentation. Fast uniFrac analysis of all the samples of different sites and dates showed that there was a similarity in the microbial community structures regardless of samples as the augmentation advanced in comparison with before- and early bioaugmentation event, indicating occurrence of changing of the indigenous microbial community structures. It was concluded that the bioaugmentation could improve the polluted water quality and simultaneously change the microbial community structures via their niche changes. This in situ remediation technology will contribute to an eco-friendly and economically cleaning up of polluted streams of brine water and freshwater.
The non-ionic surfactant (NIS) Tween 80 was evaluated for its ability to influence invitro cumulative gas production, dry matter digestibility, cellulolytic enzyme activities, anaerobic microbial growth rates, and adhesion to substrates by mixed rumen microorganisms on rice straw, alfalfa hay, cellulose filter paper and tall fescue hay. The addition of NIS Tween 80 at a level of 0.05% increased significantly (P<0.05) in vitro DM digestibility, cumulative gas production, microbial growth rate and cellulolytic enzyme activity from all of substrates used in this study. In vitro cumulative gas production from the NIS-treated substrates; rice straw, alfalfa hay, filter paper and tall fescue hay was significantly (P<0.05) improved by 274.8, 235.2, 231.1 and 719.5% compared with the control, when substrates were incubated for 48 hr in vitro. The addition of 0.05% NIS Tween 80 to cultures growing on alfalfa hay resulted in a significant increase in CMCase (38.1%), xylanase (121.4%), Avicelase (not changed) and amylase (38.2%) activities after 36 h incubation. These results indicated that the addition of 0.05% Tween 80 could greatly stimulate the release of some kinds of cellulolytic enzymes without decreasing cell growth rate in contrast to trends reported with aerobic microorganism. Our SEM observation showed that NIS Tween. 80 did not influence the microbial adhesion to substrates used in the study. Present data clearly show that improved gas production, DM digestibility and cellulolytic enzyme activity by Tween 80 is not due to increased bacterial adhesion on the substrates.
By using Korean native soybean, traditional meju was prepared in Chuncheon, Kangweondo according to the traditional process. Analysis of physico-chemical, enzymatic and microbiological changes during meju fermentation were carried out in order to obtain a basic information for industrial scale production of meju. The enviroments for natural meju fermentation were $10{\sim}15^{\circ}C$ and $60{\sim}70%{\;}RH$. Moisture content decreased from 59% to 11% (exterior section) and 19% (interior section). the pH of meju rapidly increased up to 8.5 at $33^{rd}{\;}day$ of fermentation and thereafter decreased down to 7.9 at $70^{th}{\;}day$ of fermentation. Souble protein content was 1.47% at initial stage and increased up to $6.31{\sim}7.34%$ at $33^{rd}{\;}day$ of fermentation. Amino nitrogen content was $460{\sim}770{\;}mg%$ at $70^{th}{\;}day$ of fermentation. the color of meju became gradually black and decreased in redness and yellowness. During the process, protease and lipase seemed to play an important role in the digestion of soy protein and fat. Acidic protease activity increased up to $135.9{\sim}152.4{\;}unit/g$ at $33^{rd}{\;}day$ of fermentation and were $181.3{\sim}272.6{\;}unit/g$ at $70^{th}{\;}day$ of fermentation. Lipase activity increased up to 6 unit/g (interior section) and 15 unit/g (exterior section) at $70^{th}{\;}day$ of fermentation. the viable cell count of meju was at the level of $10^8{\;}CFU/g$ during the overall fermentation period. Aerobic halophilic count was $1.51{\times}10^7{\;}CFU/g$ at initial stage and maintained $10^8{\;}CFU/g$ level during the process. Initial anaerobic cell count was $2.0^9{\times}10^4{\;}CFU/g$ and increased up to $10^5{\;}CFU/g$ level at 47 days. Yeast and mold counts were $10^4{\sim}10^5{\;}CFU/g$ for the fermentation period.
This study assessed hazards at the harvest stage of strawberry farms which may cause risk to humans. A total of 216 samples were collected from 6 strawberry farms (soil culture farms: A, B, C; nutriculture farms: D, E, F) located in Western Gyeongnam. The collected samples were subjected for sanitary indicator bacteria (aerobic plate count, coliforms and Escherichia coli), major foodborne pathogens (E. coli O157:H7, Listeria monocytogenes, Salmonella spp., Staphylococcus aureus and Bacillus cereus), and fungi. The levels of APC and coliform in the soil culture farms were 1.0-6.9 and 0.4-4.6 log CFU/g (leaf, mL, hand or 100 $cm^2$), respectively. The samples obtained from the nutriculture farms were contaminated with the levels of 0.8-4.9, and 0.2-2.6 log CFU/g (leaf, mL, hand or 100 $cm^2$) of APC and coliform. However, E. coli was not detected in any samples. In major foodborne pathogens, S. aureus was detected at the level of ${\leq}$3.3 log CFU/hand in workers' hand samples and B. cereus was detected at the levels of 0.4-4.1 log CFU/g (hand or 100 $cm^2$) in soil, plants and workers' hygiene. L. monocytogenes, E. coli O157:H7 and Salmonella spp. were not detected. Fungi were detected at the levels of 1.0-5.2 and 0.2-4.4 log CFU/g (leaf, mL, hand or 100 $cm^2$) in soil culture and nutriculture farms, respectively.
Kim, Se-Ri;Kim, Jin-Bae;Lee, Hyo-Sup;Lee, Eun-Sun;Kim, Won-Il;Ryu, Song-Hee;Ha, Jihyung;Kim, Hwang-Yong;Ryu, Jae-Gee
Journal of Food Hygiene and Safety
/
v.30
no.3
/
pp.249-257
/
2015
The purposes of this study were to develop a small scale post-harvest facility, and consequently to evaluate the effects of applying the facility along with hygiene education on the level of microbial safety in Korean leeks production. A total of 135 samples were collected at three Korean leeks farms in Yangju, Gyeonggi province. Food safety indicators (Aerobic plate count (APC), coliform count, and Escherichia coli) and foodborne pathogens (E. coli O157:H7, Salmonella spp., Staphylococcus aureus, Listeria monocytogenes, and Bacillus cereus) on/in the samples were assessed. The microbial load measured as APC with harvesting tools such as comb, chopping board, and knife, at the farms where the small scale post-harvest facility had been operated (Farms A and B) was lower than that at another farm having no post-harvest facility (Farm C) by 1.44~2.33 log CFU / $100cm^2$. Moreover, the chopping board from Farm C was observed being contaminated with B. cereus at 6.03 log CFU / $100cm^2$. The coliform counts from the samples increased by 0.57~1.89 log CFU/g after leeks was submerged in ground water for washing. E. coli was recovered from leeks, soil, and the ground water used in the washing process, while no E. coli O157:H7, Salmonella spp., and L. monocytogenes was detected. Our results indicated that the small scale post-harvest facility developed in this study as well as the hygiene education played an important role in enhancing the level of microbial food safety in the leeks production environment. However, a disinfection technique could be needed during the washing step in order to prevent a potential contamination.
Cheon, Jin-Young;Yang, Ji Hye;Kim, Min Jeong;Lee, Su-Mi;Cha, Myeonghwa;Park, Ki-Hwan;Ryu, Kyung
Journal of Food Hygiene and Safety
/
v.27
no.4
/
pp.420-426
/
2012
The purpose of this study was to identify control points through microbiological hazard analysis in the manufacturing processes of starch noodles. Samples were collected from the ingredients, manufacturing processes, equipment and environment. Microbiological hazard assessments were performed using aerobic plate counts (APC), Enterobacteriaceae (EB), E. coli and five pathogens including B. cereus, E. coli O157:H7, L. monocytogenes, Salmonella spp., and S. aureus. The APC levels in raw materials were from 2.12 to 3.83 log CFU/g. The contamination levels after kneading were 4.31 log CFU/g for APCs and 2.88 log CFU/g for EB counts. APCs decreased to 1.63 log CFU/g and EB were not detected after gelatinization, but their levels slightly increased upon cooling, cutting, ripening, freezing, thawing, and separating. The reuse of cooling and coating water would be a critical source of microbial increase after cooling. After drying, APCs and EB counts decreased to 5.05 log CFU/g and 2.74 log CFU/g, respectively, and the levels were maintained to final products. These results suggest that the cooling process is a critical control point for microbiological safety, and the cooling water should be treated and controlled to prevent cross contamination by pre-requisite program.
To evaluate microbiological and aflatoxin safety on traditional dried persimmon, a total of 315 samples were collected from 105 farms. The collected samples were assessed on aflatoxin and microorganisms (Aerobic plate count, coliform count, Escherichia coli, Escherichia coli O157:H7, Salmonella spp., Staphylococcus aureus, Listeria monocytogenes, Bacillus cereus). The the APC of sliced dried persimmon, dried persimmon, and semi dried persimmon were $3.93{\pm}0.96$, $2.12{\pm}0.93$, and $1.50{\pm}1.08{\log}\;CFU/g$, respectively. S. aureus was detected in 40.0% of sliced dried persimmon, 29.5% of dried persimmon, and 23.5% of semi dried persimmon. E. coli recovered from dried persimmon and semi dried persimmon was 6.6%, and 2.9%, respectively. However, E. coli O157:H7, Salmonella spp., and L. monocytogenes were not detected. According to the result of aflatoxin by ELISA and UPLC, aflatoxin was not detected in any sample. These data suggested that safety management system should be introduce to the farms producing traditional dried persimmon to enhance the safety of traditional dried persimmon.
Kim, Sol-A;Lee, Jeong-Eun;Kim, Go-Un;Kim, Soo-Hwan;Shim, Won-Bo
Journal of Food Hygiene and Safety
/
v.32
no.6
/
pp.485-492
/
2017
This study investigated to determine the microbial contamination levels of strawberries at harvest and distribution stages and to suggest a control measure for reducing the microbial contamination of strawberries by replacing worker's gloves used at harvest and distribution stages. According to the monitoring results, the contamination levels of total aerobic bacteria (TAB) were in the order of soil ($7.12{\pm}0.61{\log}_{10}CFU/g$), gloves ($6.06{\pm}1.80{\log}_{10}CFU/cm^2$), strawberry ($3.28{\times}0.98{\log}_{10}CFU/g$), and water ($3.08{\pm}0.55{\log}_{10}CFU/g$) at harvest stage. TAB of strawberry at was harvest stage reduced from $3.28{\pm}0.98{\log}_{10}CFU/g$ to $1.85{\pm}0.21{\log}_{10}CFU/g$ and $2.6{\pm}0.30{\log}_{10}CFU/g$ at cold and room temperature storage, respectively. By the replacement of worker's gloves and distribution temperature, TAB levels of the strawberries were significantly reduced when compared to those of the strawberries treated without replacement of worker's gloves and distributed at room temperature. For reusing the replaced gloves, washing with a commercial disinfectant, clorox, was effective to reduce microorganisms contaminated on the worker's gloves. These results demonstrated that appropriate replacement of gloves at the harvest and distribution stages is an effective method for reducing microbial contamination of fresh strawberries.
The study analyzed the effects of salt concentration [high salt (HS) and low salt (LS)] and sodium nitrite ($NaNO_2$), which are typically utilized in Korean processing facilities, on fatty acid composition, free amino acids, microbial counts and sensory characteristics of processed dry-cured ham. Four different treatments were considered: three hams (11.30 kg) salted with 92 g/kg salt (w/w) (HS), three hams (10.65 kg) treated with HS and 100 ppm $NaNO_2$ (HS+$NaNO_2$), three hams (11.42 kg) salted with 62 g/kg salt (w/w) (LS), and three hams (10.62 kg) treated with LS and 100 ppm $NaNO_2$ (LS+$NaNO_2$). Fatty acid composition analysis revealed significantly (p<0.05) higher saturated fatty acid and lower (p<0.05) unsaturated fatty acid in the HS+$NaNO_2$ group compared with the other groups. Glutamate, alanine and lysine free amino acids were higher than the other free amino acids. The processing conditions did not significantly affect the free amino acids of biceps femoris muscles, except for the proline content (p>0.05). In sensory evaluation, the fermentation aroma of the LS group was higher than that of the HS group. The aerobic counts consistently ranged from from $2.3{\times}10^2$ to $1.11{\times}10^4$ CFU/g. Escherichia coli including strain O157:H7, Staphylococcus aureus, and Salmonella spp. were not detected.
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