• Journal of Life Science
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• v.26 no.12
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• pp.1376-1382
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• 2016

Xylitol is widely used in the food and medical industry. It is produced by the reduction of xylose (lignocellulosic biomass) in the Saccharomyces cerevisiae strain, which is considered genetically safe. In this study, the expression system of the GRE3 (YHR104W) gene that encodes xylose reductase was constructed to efficiently produce xylitol in the S. cerevisiae strain, and the secretory production of xylose reductase was investigated. To select a suitable promoter for the expression of the GRE3 gene, pGMF-GRE3 and pAMF-GRE3 plasmid with GAL10 promoter and ADH1 promoter, respectively, were constructed. The mating factor ${\alpha}$ ($MF{\alpha}$) signal sequence was also connected to each promoter for secretory production. Each plasmid was transformed into S. cerevisiae $SEY2102{\Delta}trp1$, and $SEY2102{\Delta}trp1$/pGMF- GRE3 and $SEY2102{\Delta}trp1$/pAMF-GRE3 transformants were selected. In the $SEY2102{\Delta}trp1$/pGMF-GRE3 strain, the total activity of xylose reductase reached 0.34 unit/mg-protein when NADPH was used as a cofactor; this activity was 1.5 fold higher than that in $SEY2102{\Delta}trp1$/pAMF-GRE3 with ADH1 as the promoter. The secretion efficiency was 91% in both strains, indicating that most of the recombinant xylose reductase was efficiently secreted in the extracellular fraction. In a baffled flask culture of the $SEY2102{\Delta}trp1$/pGMF-GRE3 strain, 12.1 g/l of xylitol was produced from 20 g/l of xylose, and ~83% of the consumed xylose was reduced to xylitol.

• Korean journal of food and cookery science
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• v.26 no.3
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• pp.335-345
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• 2010

The objective of this study was to model the kinetics of S. aureus survival on high risk foods in school foodservice operations. After inoculating S. aureus ATCC25923 onto the various high risk foods, the effects of competitive microorganism, storage temperatures($25^{\circ}C$, $35^{\circ}C$), and initial contamination levels ($1.0{\times}10^2\;CFU/g$, $1.0{\times}10^5\;CFU/g$) on the growth of S. aureus were investigated. Lag time decreased and specific growth rate increased with a storage temperature ($25^{\circ}C$<$35^{\circ}C$) and with a higher initial inoculation level ($1.0{\times}10^2\;CFU/g$<$1.0{\times}10^5\;CFU/g$). Previously it was shown that S. aureus is a weaker competitor than other organisms, but it proliferates aggressively in a noncompetitive environment. However, in our study, when S. aureus was used to inoculate japchae (glass noodles with sauteed vegetables) and meat ball, the growth of S. aureus was similar and more active with competitive organisms than that without competitive organisms. Regardless of other factors, the initial level of S. aureus was a more significant factor of the growth. High inoculation levels of S. aureus were reached at 6 log CFU/g within 3 hours. An incubation temperature of $35^{\circ}C$ and the animal protein component of menu items also were identified as significant factors influencing the growth of S. aureus. Therefore, the duration of time meals are stored before serving should be considered a critical control point. Food service providers must control time and temperature to insure the safety of cooked foods.

• Korean Journal of Pharmacognosy
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• v.5 no.2
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• pp.111-124
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• 1974

The radioactive compound sodium $acetate-U-C^{14}\;(C^{14}-acetate)$ was administered to two- and four-year-old July and September American ginseng (Araliaceae, Panax quinquefolium L.) plants and cuttings. The $C^{14}-acetate$ uptake was approximately 99%. The autoradiochromatograms suggest that the saponins isolated by preparative thin-layer chromatography contained impurities, especially those isolated from the leaf and stem extracts. The root and fruit methanol extracts yielded relatively pure saponins. The large amounts of panaquilin B and its proximity to panaquilin C on preparative thin-layer plates resulted in some admixing. The average concentration (% plant dry weight) of semi-purified saponins were high in the leaves (13.8%), as compared to fruits (9.8%), stems (7.9%) and roots (6.3%). The average percentage of $C^{14}-acetate$ incorporation into panaquilins was 4.8%. The average percentage of $C^{14}-acetate$ incorporation into panaquilins B and C was higher (1.40% and 1.13%, respectively) than that into panaquilins C, (d), G-1 and G-2 (0.75%, 0.65%, 0.13% and 0.53%, respectively). Panaquilin synthesis may be depending upon the part, collection period and age of the plant. The average percentage of $C^{14}-acetate$ incorporation into panaquilin B is high in roots (0.58%) and stems (0.48%); that into panaquilins C and (d) high in leaves (0.40% and 0.45%, respectively); and that into panaquilin E high in roots and leaves (0.55% and 0.50%, respectively). Panaquilin G-2 was synthesized in all parts of plants. The panaquilins appear to be biosynthesized more actively in July than September (exception-panaquilin G-1). Panaquilins B, C and G-1 may be biosynthesized more actively in four-year-old plants and panaquilins (d) and E more actively in two-year-old plants. The results from expectance with cuttings suggest that the panaquilins are synthesized de novo in the above-ground parts of ginseng plants, and that panaquilin G-1 may be synthesized de novo in the leaf. It is known from the tissue culture studies that panaquilins are produced by leaf, stem and root callus tissues and cailus-root cultures of American and Korean ginseng plants. Panaquilins may actively be synthesized de novo in most any cell or organ of the ginseng plants. It was verified that $C^{14}-acetate$ was incorporated into the panaxadiol portions of the panaquilins of two-year-old plants (sp. act. 0.56 mmcCi/mg) and four-year-old plants $(sp.\;act.\;0.54\;m{\mu}Ci/mg)$.

• Research in Plant Disease
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• v.15 no.2
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• pp.120-126
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• 2009

In the course of study on the roles of phytotoxins in the pathogenicity of Botrytis cinerea, we isolated two novel phytotoxins. They were identified as 3-O-acetyl botcinol and 3-O-acetyl botcinolide. In this study, we investigated correlation between the two phytotoxins and the pathogenicity of B. cinerea. In liquid cultures, the two phytotoxins were not produced by three low pathogenic isolates out of 25 B. cinerea isolates. Among strong or moderate pathogenic isolates, some produced the two phytotoxins, but the others did not. On the other hand, the ethyl acetate extracts of fermentation broths of 10 out of 25 isolates showed phytotoxic activity against various plants tested in a whole plant assay. The phytotoxins were detected in all of the 10 phytotoxic ethyl acetate extracts. In planta, the two phytotoxins were detected in all of the plant tissues infected with strong pathogenic isolates. However, there was no correlation between the ability of B. cinerea isolates to produce the two phytotoxins and their pathogenicities. The two phytotoxins began to detect in tomato plant tissues infected with B. cinerea 2-16 at 3 days after inoculation, increased gradually till 4 days after inoculation, and then decreased. The above results suggest that 3-O-acetyl botcinol and 3-O-acetyl botcinolide are one of pathogenicity factors for B. cinerea, but not a primary determinant of its pathogenicity.

• Weed & Turfgrass Science
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• v.6 no.3
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• pp.222-234
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• 2017

Green tides, which was mainly caused by Ulva spp., have been increasing in severity and frequency globally, and have negatively affected on marine ecosystems. This study was conducted to investigate effects of various physical and chemical factors on the death of Ulva australis (ULAUS) and to consider a practical measures useful for alleviating Ulva bloom. Soaking of ULAUS thalli in pure water for 8 hr didn't induce a death, but incubation in 1.0-1.5% salinity for 7 d inhibited sporulation by about 70%. Desiccation gave rise to a serious damage when more than 40-50% of initial fresh weight was lost. ULAUS growth was sensitive to temperature and seriously inhibited from more than $30^{\circ}C$. At $35^{\circ}C$, $40^{\circ}C$, $45^{\circ}C$ and $50^{\circ}C$, treatment time required for 90-95% death of ULAUS thalli was 1 d, 10 min, 30 sec, and 1 sec, repectively. ULAUS growth was seriously inhibited at lower than pH 6.0 and completely dead at pH 4.0. Several compounds for ULAUS control was selected and the chemcals causing a rapid death were oxidants such as hydrogen peroxide and sodium percarbonate. Taken together, our results suggest that low salinities, dryness, pH, high temp. and compounds could be selected for Ulva bloom control, and high temperature and compounds seems to be useful for a development of practical control methods.

• Korean Journal of Microbiology
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• v.40 no.4
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• pp.275-281
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• 2004

Our previous research has demonstrated that the bacterium, Pseudomonas sp. NFQ-l capable of utilizing quin­oline (2,3-benzopyridine) as the sole source of carbon, nitrogen, and energy was isolated and characterized [Yoon et ai. (2003) Kor. J. Biotechnol. Bioeng. 18(3):174-179]. In this study, we have found that Pseudomonas sp. NFQ-l could degrade quinoline as well as benzoate, and extended this work to characterize the catechol 1,2­dioxygenase (C1,2O) purified from the bacterium cultured in benzoate media. Initially, C1,2O has been purified by ammonium sulfate precipitation, gel permeation chromatography, and Source 15Q. After Source 15Q, puri­fication fold was increased to approximately 14.21 unit/mg. Molecular weight of C1,2O was about 33 kDa. Physicochemical characteristics (e.g., substrate specificity, Km, Vmax, pH, temperature and effect of inhibitors) of purified C1,2O were examined. C1,2O demonstrated the activity for catechol, 4-methylcatechol and 3-meth­ylcatechol as a substrate, respectively. The Km and Vmax value of C1,2O for catechol was 38.54 ${\mu}M$ and $25.10\;{\mu}mol{\cdot}min^{-1}{\cdot}mg^{-1}.$ The optimal temperature of C1,2O was $30^{\circ}C$ and the optimal pH was approximately 8.5. Metal ions such as $Ag^+,\;Hg^+,\;Ca^{2+},\;and\;Cu^{2+}$ show the inhibitory effect on the activity of C1,2O. N-terminal amino sequence of C1,2O was analyzed as ^1TVKISQSASIQKFFEEA^{17}.$ In this work, we found that the amino acid sequence of NFQ-l showed the sequence homology of 82, 71, 59 and $53\%$ compared with C1,2O from Pseudomonas aeruginosa PA0l, Pseudomonas arvilla C-1., P. putida KT2440 and Pseudomonas sp. CA10, respectively.

• Journal of Animal Science and Technology
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• v.45 no.1
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• pp.131-142
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• 2003

We evaluated the effects of adding fibrolytic enzyme into ruminant diets on ruminal fermentation (in vitro) and lactational performances of dairy cows (in vivo). Through the in vitro experiment that was carried out with different contents of NDF (34, 38, 43%) in diets, digestibilities of NDF in the rumen appeared not significantly different by the addition of enzyme but were different by NDF content in diets showing higher digestibility in NDF 43% diet. It could be attributed by the relatively higher amount of hemicellulose in the current experimental diets than in conventional diets that might have been digested easily by the addition of fibrolytic enzyme in the rumen. The addition of fibrolytic enzyme tended to increase NDF digestibilities to a little extent both in 0.05 and 0.1% enzyme levels. Ruminal pH, NH3-N concentrations and VFA production in the rumen were not affected by the addition of fibrolytic enzyme. Activities of CMCase and xylanase were higher in enzyme treated diets of both NDF 34 and 38%. In particular, the activities of xylanase that slowly decreased from 0 to 12 hr but rapidly after 24 hr indicates that the major action of the enzyme in the rumen occurs in early period of incubation. Through an in vivo experiment, fibrolytic enzyme addition into the diets of dairy cows indeed affected lactational performance of milk yield. The cows fed enzyme treated diets produced 8% (1.9kg/d) more amounts of milk than with no enzyme addition. Milk composition of milk fat and protein was not affected by enzyme addition. Overall, the results of this in vivo study indicates that fibrolytic enzyme can be used to improve milk production in lactating cows. In respect that animals in different treatments of this study had the same amounts of intake, the increased milk yield with enzyme addition may be attributed to the improved utilization of nutrients in the digestive tract.

• Applied Microscopy
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• v.36 no.3
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• pp.165-172
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• 2006

Secretory leukocyte protease inhibitor (SLPI) involves tissue protection against the destructive action of neutrophil elastase at the site of inflammation. Several studies on new functions of SLPI have demonstrated that SLPI may play a primary role in innate immunity than protease inhibitor, To identify the function of SLPI by lipopolysaccharide (LPS) stimulation in the embryonic fibroblast (NIH3T3) cells. we studied the expression of SLPI compared to other growth factors involving the LPS treatment. To address this, we performed the reverse transcriptase polymerase chain reaction (RT-PCR) and Western blots for the detection of mRNA and protein expression of the SLPI and some growth factors such as VEGF. bFGF, and PDGF-BB after LPS stimulation. NIH3T3 cells were exposed 100 ng/mL Escherichia coli LPS for 30min, 60min, 90min, 24h, and 48h, respectively. The result of RT-PCR showed that SLPI and VEGF mRNA was expressed strongly in NIH3T3 without related to LPS stimulation. mRNA of bFGF was weakly expressed such as the expression of the control. PDGF mRNA expression gradually increased follows at time course. However, SLPI protein level was increased in lysates and culture medium by LPS stimulation. Phase contrast microscopic and scanning electron microscopic observation showed that the increased cell number and cytoplasmic enlargement of the NIH3T3 cells. Therefore, it suggests that the LPS upregulates SLPI expression in NIH3T3 cells. Moreover, secreted SLPI may stimulate cell proliferation and migration.

• Journal of Animal Science and Technology
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• v.48 no.5
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• pp.699-708
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• 2006

Ground rice, barley and corn were fed separately to the ruminally cannulated Hanwoo (Korean native cattle) for comparing their in situ and in vitro degradabilities, microbial growth, pH and gas production. It has been found that nearly all the dry matter (DM) and organic matter (OM) in barley and rice disappeared during 24 hr suspension in the rumen, but those in corn were only reduced by around 67%. Water soluble DM and OM fractions(‘a’), ranked from highest to lowest was corn, then rice and finally barley, but the order was reversed for content ‘b’, degradable fraction during time ‘t’. Judging by the degradation parameter of ‘b’ fraction, degradation rates per hour of DM and OM for barley were 38.3% and 37.2% respectively, significantly higher than those for rice (7.7% and 5.6%) and corn (4.1% and 1.3%). In general, results obtained from in vitro degradability of DM and OM were lower than those from in situ trials, but the ranking order of degradability was in agreement between both trials. In particular, ground rice has relatively lower in vitro microbial growth than corn or barley, but exhibited higher gas production. In addition, in vitro microbial growth of ground rice increased with up to 12 hr of incubation period, thereafter experienced a decrease with extended incubation time. pH of in vitro solution of rice decreased following 9 hr of incubation but gas production increased rapidly during the same period. From the results of DM and OM degradabilities and pH changes of in vitro solution with incubation time, it is concluded that rice represents a good source of energy for stability of rumen fermentation.

• Horticultural Science & Technology
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• v.32 no.5
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• pp.628-635
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• 2014

The large amount of $CO_2$ emitted from mushrooms during incubation and developmental stages can be utilized in plant production systems as a $CO_2$ source. The objectives of this study were to measure the $CO_2$ emission and absorption rates of mushroom and lettuce, respectively, and to analyze the $CO_2$ concentrations at various ratios of mushroom and lettuce in a closed production system. The $CO_2$ emission rate of king oyster mushrooms (Pleurotus eryngii ( DC.) Qu$\acute{e}$l) and $CO_2$ absorption rate of lettuces (Lactuca sativa L. cv. Asia Heuk Romaine) were measured by using two closed acryl chambers ($1.0m{\times}0.8m{\times}0.5m$) in which indoor temperatures were maintained at $18^{\circ}C$ and $22^{\circ}C$, respectively. The lettuce was grown at a light intensity of PPF $340mol{\cdot}m^{-2}{\cdot}s^{-1}$ and with nutrient solution at EC $1.2dS{\cdot}m^{-1}$. The air was periodically circulated between the two chambers using a diaphragm pump. The $CO_2$ emission rate of the mushroom increased until the $15^{th}$ day after scratching (DAS) and then decreased. The rate also increased with increased indoor temperature. In particular, the $CO_2$ emission rate per fresh weight of fruit body increased by about 3.1 times after thinning compared to before thinning. In terms of $CO_2$ balance, the $CO_2$ emission rates from a bottle (950 mL) of the mushroom at 9, 12, and 14 DAS were equivalent to those of 3, 4.5, and 5.5 lettuce plants at 7, 10, and 12 DAT (days after transplanting), respectively. This work shows that balance in $CO_2$ concentration could be achieved using an appropriate ratio of the two crops in a closed production system.