L-Xylosone was detected as its quinoxaline derivative in the degradation solution of dehydro-L-ascorbic acid. The production rate of L-xylosone was much faster in aerated phosphate-cirate buffer (pH 5. 6) than in pure water. L-Xylosone and dehydro-L-ascorbic acid were identified in the crude extracts of Saccharomyces cerevisiae. The concentration of L-xylosone in the crude cell extracts was calculated to be about 0.2 nmol $(mg protein)^{-1}$. When L-xylosone was added to asynchronous culture of S. cerevisiae, it inhibited primarily the synthesis of protein and RNA. Examination of the effect of L- xylosone on synchronous culture of the yeast indicated that L-xylosone inhibited the initiation of DNA replication and that the cells were arrested at $G_1$, stage of cell division cycle. These results suggested a possibility that L-xylosone can act as an inhibitor of cell growth.
The objective of this study was to evaluate the antimicrobial effects of various natural flavonoids against growth of psychotropic Bacillus cereus strains, which cause dairy food outbreaks. Flavonoids were first screened for their ability to inhibit growth of B. cereus strains using the paper-disc diffusion test. Second, the growth inhibitory effect of selected flavonoids was evaluated in tryptic soy broth supplemented with 0.6% yeast extract, and the bactericidal effect of the flavonoids was measured in 0.8% (w/v) NaCl solution. Based on the paper-disc diffusion test, kaempferol was effectively active against B. cereus P14 and B. cereus KCCM 40935. Kaempferol had an antimicrobial effect at concentrations greater than 100 ${\mu}M$, and the numbers of B. cereus P14 and B. cereus KCCM 40935 decreased by 3.55 and 1.5 log cycles, respectively. The cell numbers of B. cereus P14 and B. cereus KCCM 40935 treated with 50 ${\mu}M$ kaempferol were reduced by 4.18 and 2.84 log cycles during a 24 h incubation to test the bactericidal effect of kaempferol (p<0.05). The results indicate that kaempferol had the greatest antimicrobial effect among the psychotropic B. cereus strains and the natural flavonoids tested.
Kim, Jennifer Jooyoun;Kwon, Young-Kyung;Kim, Ji Hyung;Heo, Soo-Jin;Lee, Youngdeuk;Lee, Su-Jin;Shim, Won-Bo;Jung, Won-Kyo;Hyun, Jung-Ho;Kwon, Kae Kyoung;Kang, Do-Hyung;Oh, Chulhong
Journal of Microbiology and Biotechnology
/
v.24
no.11
/
pp.1559-1565
/
2014
Cellulase and xylanase are main hydrolysis enzymes for the degradation of cellulosic and hemicellulosic biomass, respectively. In this study, our aim was to develop and test the efficacy of a rapid, high-throughput method to screen hydrolytic-enzyme-producing microbes. To accomplish this, we modified the 3,5-dinitrosalicylic acid (DNS) method for microwell plate-based screening. Targeted microbial samples were initially cultured on agar plates with both cellulose and xylan as substrates. Then, isolated colonies were subcultured in broth media containing yeast extract and either cellulose or xylan. The supernatants of the culture broth were tested with our modified DNS screening method in a 96-microwell plate, with a $200{\mu}l$ total reaction volume. In addition, the stability and reliability of glucose and xylose standards, which were used to determine the enzymatic activity, were studied at $100^{\circ}C$ for different time intervals in a dry oven. It was concluded that the minimum incubation time required for stable color development of the standard solution is 20 min. With this technique, we successfully screened 21 and 31 cellulase- and xylanase-producing strains, respectively, in a single experimental trial. Among the identified strains, 19 showed both cellulose and xylan hydrolyzing activities. These microbes can be applied to bioethanol production from cellulosic and hemicellulosic biomass.
In this study, an attempt was made to increase the survival rate of bifidobacteria entrapped in alginate in the gastrointestinal tract, and to investigate the potential industrial applications, for example lyophilized capsules and yogurt. First, the protective effect of various food additives on bifidobacterial survivability was determined after exposure to simulated gastric juices and bile salts. The additives used in this study were skim milk (SM), polydextrose (PD), soy fiber (SF), yeast extract (YE), chitosan (CS), $\kappa$-carageenan ($\kappa$-C) and whey, which were added at 0.6% concentration (w/v) to 3% alginate-bifidobacterial solution. In the simulated gastric juices and bile salts, the protective effect of 0.6% skim milk-3% alginate (SM-A) beads on the survival rate of bifidobacteria proved to be higher than the other additives. Second, the hydrogen ion permeation was detected through SM-A vessel without bifidobacterial cells at different SM concentrations (0.2%, 0.4%, 0.6%, 0.8%, and 1.0%). There were no differences in terms of the pH decrease in SM-A vessels at 0.6%, 0.8%, and 1.0% (w/v) SM concentrations. The survival rate of bifidobacteria in SM-A beads would appear to be related to the SM buffering capacity against hydrogen ions and its tendency to reduce the pore size of bead. In this experiment, the survival rate of bifidobacteria entrapped in beads containing 0.6% SM showed the highest viability after exposure to simulated gastric juices for 3h, thereby indicating that 0.6% SM is the optimum concentration fir 3% alginate bead preparation. Third, the effect of SM-A beads on the freeze-drying and yogurt storage for 10 days was investigated. SM-A beads were found to be more efficient for freeze drying and yogurt storage than untrapped cells and the alginate bead. Consequently, the survival rate of bifidobacteria entrapped in SM-A beads was increased in simulated gastric juices, bile salts and probiotic products, such as lyophilized capsules and yogurt, SM-A beads can be expected to produce high value probiotic products.
Deoxynivalenol (DON)-contaminated barley was treated with 0.1 M $Na_2CO_3$ solution to reduce DON content. DON content of barley was reduced from 2.08 to 0.67 ppm. Bread was made with 10, 20, and 30% DON-reduced barley flour added to white wheat flour. Farinogram showed water absorption and arrival time increased, while stability and mechanical tolerance index decreased when DON-reduced barley flour was added to white wheat flour. Gelatinization temperature, temperature at maximum viscosity, and maximum viscosity increased in amylogram with increasing addition of DON-reduced barley flour. Loaf volume of bread decreased with increasing amount of DON-reduced barley flour, while loaf weight increased. Barley flour pH increased by treatment with$Na_2CO_3$, and pH reduction decreased fermentation rate of yeast. Volume and size of gluten matrix decreased and crumb hardened in bread made with DON-reduced barley flour. Acceptabilities for color and texture were low in bread made with DON-reduced barley flour. Addition of DON-reduced barley flour at 30% reduced overall acceptability, whereas no significant difference in overall acceptability was observed when DON-reduced barley flour was added at 10 and 20%.
Secretion of the expressed heterologous proteins can reduce the stress to the host cells and is beneficial to their recovery and purification. In this study, fed-batch cultures of Escherichia coli YK537 (pAET-8) were conducted in a 5-L fermentor for the secretory production of human epidermal growth factor (hEGF) whose expression was under the control of alkaline phosphatase promoter. The effects of feeding of glucose and complex nitrogen sources on hEGF production were investigated. When the fed-batch culture was conducted in a chemically de-fined medium, the cell density was 9.68 g/L and the secreted hEGF was 44.7 mg/L in a period of 60 h. When a complex medium was used and glucose was added in pH-stat mode, the secreted hEGF was improved to 345 mg/L. When the culture was fed with glucose at a constant specific rate of $0.25\;gg^{-1}h^{-1}$, hEGF reached 514 mg/L. The effects of adding a solution containing yeast extract and tryptone were further studied. Different rate of the nitrogen source feeding resulted in different levels of phosphate and acetic acid formation, thus affected hEGF expression. At the optimal feeding rate, hEGF production achieved 686 mg/L.
The effects of phosphate alternatives on meat quality in marinated chicken were investigated with the application of chilling and freezing. Breast muscles were injected with solution of the green weight containing 1.5% NaCl and 2% sodium tripolyphosphate (STPP) or phosphate alternatives. Treatment variables consisted of no phosphate [control (-)], 0.3% sodium tripolyphosphate [control (+)], 0.3% prune juice (PJ), 0.3% oyster shell, 0.3% nano-oyster shell, and 0.3% yeast and lemon extract (YLE) powder. One-third of the meat samples were stored at 4℃ for 1 d, and the rest of the meats were kept at -18℃ for 7 d. In chilled meat, a lower drip loss was noted for control (+) and YLE, whereas higher cooking yield in YLE compared to all tested groups. Compared with control (+), the other treatments except PJ showed higher pH, water holding capacity, moisture content, lower thawing and cooking loss, and shear force. Natural phosphate alternatives except for PJ, improved the CIE L* compared to control (-), and upregulated total protein solubility. However, phosphate alternatives showed similar or higher oxidative stability and impedance measurement compared to control (+), and an extensive effect on myofibrillar fragmentation index. A limited effect was observed for C*, h°, and free amino acids in treated meat. Eventually, the texture profile attributes in cooked of phosphate alternatives improved except for PJ. The results indicate the high potential use of natural additives could be promising and effective methods for replacing synthetic phosphate in chilled and frozen chicken with quality enhancement.
For the effect of water-soluble degraded chitosan on the shelf-life of tofu, sterilized distilled water, 0.5% degraded chitosan, 0.5% fumaric acid and 0.5% lactic acid used as an tofu-immersion solutions were investigated by microbial counts, pH, and turbidity during the periods of storage at $4^{\circ}C$. After 2 weeks storage, total aerobic microbial counts in tap water and sterilized distilled water used as an immersion solution were $3.8\;{\times}\;10^8$ and $1.8\;{\times}\;10^8\;CFU/mL$, respectively. In 0.5% fumaric acid and 0.5% lactic acid immersion solutions, the microbial counts were around $10^7\;CFU/mL$ after 2 weeks while the microbial population in 0.5% water-soluble degraded chitosan were, however, $1.6\;{\times}\;10^5\;CFU/mL$ after 2 weeks and $1.7{\times}10^7\;CFU/mL$ after 3 weeks. The lag phase of initial contaminated microbes in 0.5% degraded chitosan solution was longer than those of other treatments. The addition of 0.5% fumaric acid and 0.5% lactic acid decreased the initial pH to pH 5.0, while those of tap water, sterilized distilled water and 0.5% degraded chitosan stabilized the immersion solution at around pH 7.2. All initial pH values were decreased during storage and then slowly increased as storage time was increased. The turbidities in all treatments were increased during storage, but the addition of 0.5% degraded chitosan showed the lowest change, compared to other treatments, showing that the water-soluble degraded chitosan has a good antimicrobial effect and has a potential use to extend the shelf-life of tofu product.
Calcium carbonate ($CaCO_3$) immobilized with alginate was studied as buffer system to enhance the cultivation efficiency of Bifidobacterium longum (ATCC 15707) which is inhibited at low pH. To test the bufferring effect of the immobilized $CaCO_3$ beads, pH value in each modified trypticase-proteose peptone-yeast (TPY) broth which is adjusted to pH 4.0 with acetic acid, lactic acid and complex solution of acetic and lactic acid, 3:2 (M:M) was tested by concentration of $CaCO_3$ bead and reaction time. The bufferring effect of $CaCO_3$ bead became higher with increasing the amount of $CaCO_3$ bead in the acidic solution. The growth rate of bifidobacteria and bufferring effect were examined in relation to the amount of $CaCO_3$ bead and concentration of glucose in the modified TPY media. The growth rate of bifidobacteria and bufferring effect were increased with increasing the amount of $CaCO_3$ bead and concentration of glucose. Also, the exponential time of bifidobacteria became longer with increasing the amount of $CaCO_3$ bead and concentration of glucose in the modified TPY media. When we observed the growth rate of bifidobacteria by the method of pH-controlled culture and $CaCO_3$ buffer system, the $CaCO_3$ buffer system was more effective than that of pH-controlled culture. Therefore, this $CaCO_3$ buffer system may be useful as a method to enhance of the cultivation efficiency of bifidobacteria.
The whole plant of crop maize was chopped and ensiled in double-layered polyethylene bags to determine the influence of residual sugar on the fermentation of lactic acid and aerobic stability by L. buchneri in whole crop maize silage made in airstress condition. There were a total of six treatments used in this experiment as follow: added 25 g de-mineralised water per kg chopped maize serving as control (con), 37.5 g glucose solution containing 12.5 g glucose ($g_1$), 75 g glucose solution containing 25 g glucose ($g_2$), 25 g, L,.buchneri suspension intended for $10^6$ cfu $g^{-1}$ (L.b.), $g_1$+L.b. and $g_2$+L.b. All silos were opened at day 91 after ensiling for measuring the pH values, microbiological enumeration, fermentative products and aerobic stability. The dry matter loss increased significantly (p<0.01) due to inclusion of sugar or L. buchneri. The lower lactic acid concentrations were observed (p<0.01) in silages inoculated with L. buchneri only or in combination with sugar addition than the correspondent uninoculated silages. Compared with control silage, ethanol production was about 3 or 6-fold higher due to addition 12.5 or 25 g glucose per kg chopped maize at ensiling. The silages added with sugar contained less acetic acid concentration (p<0.01) than control, but silages inoculated with L. buchneri showed the contrary effects (p<0.01) at different sugar levels. No butyric acid was found in uninoculated silages, silages inoculated with L. buchneri. producted more propionic acid, 1-propanol and butyric acid. Lactic acid bacteria counts increased markedly (p<0.01) due to inoculation with L. buchneri, whereas it was reduced (p<0.01) by added sugar. No significant difference was observed in count of yeast, but inoculation with L. buchneri shows a decreasing trend. Mould count in all silages was less than 2 (log cfu $g^{-1}$). The added sugar had negative effects on aerobic stability of maize silage made under air-stress conditions, whereas inoculation with L. buchneri improves (p<0.01) the aerobic stability.
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