• Applied Biological Chemistry
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• v.10
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• pp.69-100
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• 1968

1. In order to investigate on the microflora and enzyme activity of mold wheat 'Nuruk' , the major source of microorganisms for the brewing of Takju (a Korean Sake), two samples of Nuruk, one prepared at the College of Agriculture, Chung Nam University (S) and the other perchased at a market (T), were taken for the study. The molds, aerobic bacteria, lactic acid bacteria, and yeasts were examined and counted. The yeasts were classified by the treatment with TTC (2, 3, 5 triphenyltetrazolium chloride) agar that yields a varied shade of color. The amylase and protease activities of Nuruk were measured. The results were as the followings. a) In the Nuruk S found were: Aspergillus oryzae group, $204{\times}10^5$; Black Aspergilli, $163{\times}10^5$; Rhizogus, $20{\times}10^5$; Penicillia, $134{\times}10^5$; Areobic bacteria, $9{\times}10^6-2{\times}10^7$; Lactic acid bacteria, $3{\times}10^4$ In the Nuruk T found were: Aspergillus oryzae group, $836{\times}10^5$; Black Aspergilli, $286{\times}10^5$; Rhizopus, $623{\times}10^5$; Penicillia, $264{\times}10^5$; Aerobic bacteria, $5{\times}10^6-9{\times}10^6$; Lactic acid bacteria, $3{\times}10^4$ b) Eighty to ninety percent of the aerobic bacteria in Nuruk S appeared to belong to Bacillus subtilis while about 70% of those in Nuruk T seemed to be spherical bacteria. In both Nuruks about 80% of lactic acid bacteria were observed as spherical ones. c) The population of yeasts in 1g. of Nuruk S was about $6{\times}10^5$, 56.5% of which were TTC pink yeasts, 16% of which were TTC red pink yeasts, 8% of which were TTC red yeasts, 19.5% of which were TTC white yeasts. In Nuruk T(1g) the number of yeasts accounted for $14{\times}10^4$ and constituted of 42% TTC pink. 21% TTC red pink 28% TTC red and 9% TTC white. d) The enzyme activity of 1g Nuruk S was: Liquefying type Amylase, $D^{40}/_{30},=256$ W.V. Saccharifying type Amylase, 43.32 A.U. Acid protease, 181 C.F.U. Alkaline protease, 240C.F.U. The enzyme activity of 1g Nuruk T was: Liquefying type Amylase $D^{40}/_{30},=32$ W.V. Saccharifying type amylase $^{30}34.92$ A.U. Acid protease, 138 C.F.U. Alkaline protease 31 C.F.U. 2. During the fermentation of 'Takju' employing the Nuruks S and T the microflora and enzyme activity throughout the brewing were observed in 12 hour intervals. TTC pink and red yeasts considered to be the major yeasts were isolated and cultured. The strains ($1{\times}10^6/ml$) were added to the mashes S and T in which pH was adjusted to 4.2 and the change of microflora was examined during the fermentation. The results were: a) The molds disappeared from each sample plot since 2 to 3 days after mashing while the population of aerobic bacteria was found to be $10{\times}10^7-35{\times}10^7/ml$ inS plots and $8.2{\times}10^7-12{\times}10^7$ in plots. Among them the coccus propagated substantially until some 30 hours elasped in the S and T plots treated with lactic acid but decreased abruptly thereafter. In the plots of SP. SR. TP. and TR the coccus had not appeared from the beginning while the bacillus showed up and down changes in number and diminished by 1/5-1/10 the original at the end stage. b) The lactic acid bacteria observed in the S plot were about $7.4{\times}10^7$ in number per ml of the mash in 24 hours and increased up to around $2{\times}10^8$ until 3-4 days since. After this period the population decreased rapidly and reached about $4{\times}10^5$ at the end, In the plot T the lactic acid becteria found were about $3{\times}10^8$ at the period of 24 fours, about $3{\times}10$ in 3 days and about $2{\times}10^5$ at the end in number. In the plots SP. SR. TP, and TR the lactic acid bacteria observed were as less as $4{\times}10^5$ at the stage of 24 hours and after this period the organisms either remained unchanged in population or ceased to exist. c) The maiority of lactic acid bacteria found in each mash were spherical and the change in number displayed a tendency in accordance with the amount of lactic acid and alcohol produced in the mash. d) The yeasts had showed a marked propagation since the period of 24 hours when the number was about $2{\times}10^8$ ㎖ mash in the plot S. $4{\times}10^8$ in 48 hours and $5-7{\times}10^8$ in the end period were observed. In the plot T the number was $4{\times}10^8$ in 24 hours and thereafter changed up and down maintaining $2-5{\times}10^8$ in the range. e) Over 90% of the yeasts found in the mashes of S and T plots were TTC pink type while both TTC red pink and TTC red types held range of $2{\times}10-3{\times}10^7$ throughout the entire fermentation. f) The population of TTC pink yeasts in the plot SP was as $5{\times}10^8$ much as that is, twice of that of S plot at the period of 24 hours. The predominance in number continued until the middle and later stages but the order of number became about the same at the end. g) Total number of the yeasts observed in the plot SR showed little difference from that of the plot SP. The TTC red yeasts added appeared considerably in the early stage but days after the change in number was about the same as that of the plot S. In the plot TR the population of TTC red yeasts was predominant over the T plot in the early stage which there was no difference between two plots there after. For this reason even in the plot w hers TTC red yeasts were added TTC pink yeasts were predominant. TTC red yeasts observed in the present experiment showed continuing growth until the later stage but the rate was low. h) In the plot TP TTC pink yeasts were found to be about $5{\times}10^8$ in number at the period of 2 days and inclined to decrease thereafter. Compared with the plot T the number of TTC pink yeasts in the plot TP was predominant until the middle stage but became at the later stage. i) The productivity of alcohol in the mash was measured. The plot where TTC pink yeasts were added showed somewhat better yield in the earely stage but at and after the middle stage the difference between the yeast-added and the intact mashes was not recognizable. And the production of alcohol was not proportional to the total number of yeasts present. j) Activity of the liquefying amylase was the highest until 12 hours after mashing, somewhat lowered once after that, and again increased around 36-48 hours after mashing. Then the activity had decreased continuously. Activity of saccharifying amylase also decreased at the period of 24 hours and then increased until 48 hours when it reached the maximum. Since, the activity had gradually decreased until 72 hours and rapidly so did thereafter. k) Activity of alkaline protease during the fermentation of mash showed a tendency to decrease continusously although somewhat irregular. Activity of acid protease increased until hours at the maximum, then decreased rapidly, and again increased, the vigor of acid protease showed better shape than that of alkaline protease throughout. 3. TTC pink yeasts that were predominant in number, two strains of TTC red pink yeasts that appeared throughout the brewing, and TTC red yeasts were identified and the physiological characters examined. The results were as described below. a) TTC pinkyeasts (B-50P) and two strains of TTC red pink yeasts (B-54 RP & B-60 RP) w ere identified as the type of Saccharomyces cerevisiae and TTC pink red yeasts CB-53 R) were as the type of Hansenula subpelliculosa. b) The fermentability of four strains above mentioned were measured as follows. Two strains of TTC red pink yeasts were the highest, TTC pink yeasts were the lowest in the fermantability. The former three strains were active in the early stage of fermentation and found to be suitable for manufacturing 'Takju' TTC red yeasts were found to play an important role in Takju brewing due to its strong ability to produce esters although its fermentability was low. c) The tolerance against nitrous acid of strains of yeast was marked. That against lactic acid was only 3% in Koji extract, and TTC red yeasts showed somewhat stronger resistance. The tolerance against alcohol of TTC pink and red pink yeasts in the Hayduck solution was 7% while that in the malt extract was 13%. However, that of TTC red yeasts was much weaker than others. Liguefying activity of gelatin by those four strains of yeast was not recognized even in 40 days. 4. Fermentability during Takju brewing was shown in the first two days as much as 70-80% of total fermentation and around 90% of fermentation proceeded in 3-4 days. The main fermentation appeared to be completed during :his period. Productivity of alcohol during Takju brewing was found to be apporximately 65% of the total amount of starch put in mashing. 5. The reason that Saccharomyces coreanuss found be Saito in the mash of Takju was not detected in the present experiment is considered due to the facts that Aspergillus oryzae has been inoculated in the mold wheat (Nuruk) since around 1930 and also that Koji has been used in Takju brewing, consequently causing they complete change in microflora in the Takju brewing. This consideration will be supported by the fact that the original flavor and taste have now been remarkably changed.

• Korean Journal of Microbiology
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• v.48 no.1
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• pp.29-36
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• 2012

This study was conducted to investigate antioxidative and physiological activities of two stage fermented Chinese Yam (TSFCY) by Monascus sp. MK2 and Lactobacillus brevis HLJ59. The extracts from TSFCY were measured to examine pigments, monacolin K contents, total polyphenol and flavonoid contents, DPPH radical scavenging activity, reducing power, ACE inhibitory activity and GABA contents. In this study, the results show that Monascus sp. MK2 and Lactobacillus brevis HLJ59, with Dioscorea batatas Dence. As the substrate can produce pigments (yellow, orange and red), monacolin K contents, total polyphenol and flavonoid contents, DPPH radical scavenging activity, reducing power, ACE inhibitory activity, and GABA contents at 14.03 (yellow), 13.88 (orange), 15.71 (red), 487.9 (MK, mg/kg), 723.8 (TP, mg/kg), 326.4 (TF, mg/kg), 81.7 (DPPH, %), and 1.5 (RP, O.D), respectively. In addition, the showed ACE inhibitory activity and GABA contents was 86.9% and 977.4 mg/kg in EtOH extract, respectively.

• Journal of The Korean Society of Grassland and Forage Science
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• v.25 no.1
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• pp.43-50
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• 2005

The purpose of this study is to discuss the ways to consistently feed high quality corn silage(CS). This study evaluated the effect of the corn silage, after a certain time has elapsed, on the chemical composition and fermentation characteristics after feedout during the winter feeding period of the CS. Six samples of CS from four dairy farms(E1, E2, E3, and L1 ) were taken in order to feed the milking cows over a winter feeding Period from November of 2002 until February of the following year, 2003(six samples were taken at the fellowing dates in the following order: sample one was taken on the 23rd Nov. 2002, sample two on the 5th of Dec. 2002, 3rd sample on the 23rd of Dec. 2002, 4th sample on the 7th of Jan. 2003, 5th sample on the 22nd of Jan. 2003, and the 6th sampling was carried out on the 6th of Feb. 2003) at the three sampling sites after the opening of the trench silos at intervals of 15 days. In the dry matter contents of CS, there wasn't any specific tendency according to the elapsed time in the range of 21.3~$27.3\%$ at all low dairy farm(E1, E2, E3, and L1). And the average dry matter contents were 24.1, 25.9, 23.6, and $20.4\%$. Considering the Proper amount of the dry matter of CS during the ripen yellow stage, the appropriate moisture content was $33\%$ (NRC, 1989), and these dry matter contents were all low. A consistent tendency was not found in the contents of CS. The average of CP contents were 10.2, 8.0, 8.5, and $9.8\%$ at the E1, E2, E3, and L1 farms, and there were significant differences. The TDN contents of CS were not different among forms according to the time elapsed. The pH, according to the time elapsed after opening of the CS, there were no differences at each of E1, E2, E3, md L1 farms. Average pH were 3.5, 3.9, 3.6, and 4.1, md all of them were in normal range. In the lactic acid contents of CS, a consistent tendency was not found among four farms. But according to the time elapsed. there was a goat difference from 1.13~$7.8\%$ The acetate, propionate, and butyrate contents of CS were very low. In this study, there was no significant difference in the CS's chemical composition and fermentation characteristics according to the elapsed time at all four dairy farms. Considering the proper dry matter contents of CS during the ripen yellow stage, the appropriate dry matter content was $33\%$, and dry matter contents of few farms were all low. To enhance the quality of corn silage should be ensiled com at proper dry matter content range from 28 to $35\%$ Therefore, content of the corn plant should be always be closely monitored prior to beginning harvest.

• Journal of The Korean Society of Grassland and Forage Science
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• v.30 no.4
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• pp.333-342
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• 2010

Corn is very important forage in Korea. The great part of them is utilized as silage. Generally, it contains a lot of grains that is feed of animal. This experiment was conducted to evaluation of fermentation ability of microbes for corn silage inoculant. Good lactic acid bacteria were isolated from good corn silage by plating MRS agar containing 0.02% sodium azide, and assessed by growing and acid producing ability in MRS broth. Six lactic acid bacteria were selected, and were found to be Gram positive, rods and catalase negative and were identified to be lactobacillus plantarum (C3-2, B13-1, CC9-1), Lactobacillus fermentum (C11-4), Lactobacillus paracasei (B14-1), and Leuconostoc lactis (A3-1) on the basis of the biochemical characteristics and utilization of substrates. Corn was ensiled at ripen stage following treatment with selected five lactic acid bacteria, two commercial inoculant, and no additive (control). After 2 month, B13-1 and CC9-1 bacteria inoculated silage were lower pH and higher lactic acid content than others treatments. The Flieg's score and grade of B13-1 and CC9-1 bacteria treated silage were higher than commercial inoculant. According to this experiment, lactobacillus plantarum B13-1 and CC9-1 strain were recommendable for good inoculant of corn silage.

• Korean Journal of Food Science and Technology
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• v.23 no.2
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• pp.123-128
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• 1991

Addition of two different salt mixtures of sodium phosphates, Ca-EDTA, $NaNO_2$ and sodium citrate were investigated for their effects on relative viscosity, textrue, sensory properties of kimchi and solids contents of kimchi and kimchi liquid during fermentation at $4{\sim}3.5^{\circ}C$. The salt mixtures were added into half fermented kimchi with the concentration range of $0.001{\sim}0.01\;M$. The results showed that higher values in viscosity of kimchi liquids were obtained for those fermented at low temperature and with salts mixtures added. The hardness of Chinese cabbage was gernerally increased until pH 4.0 reached and then decreased thereafter for those fermented without salts mixture. However the salts added kimchi showed no decrease and a slightly harder texture measured at the late stage of fermentation. Soluble solids concentration steadly decreased in kimchi liquids for those salts mixture added while those without salts mixture were initially increased followed by slow decrease. Comparison of sensory properties showed that the degree of changes was reduced when salt mixture was added. Higher scores in fresh-sourness and acidic taste, hardness and chewiness in texture and lower moldy odor were obtained when the data was compared for those kimchi having the pH range of $4.0{\sim}4.2$.

• Applied Biological Chemistry
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• v.8
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• pp.11-20
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• 1967

In order to study the change of nitrogeneous compounds during the "Natto" preparation, the contents of insoluble protein, water soluble protein and amino nitrogen were determined and the pattern of peptides and amino acids was investigated by paper chromatography for the fractions resulting from molecular sieving. The results are summarized as follows: 1. Insoluble protein nitrogen which was increased to 84% by autoclaving the native soybean decreased to 44%, whereas the trichloroacetic acid soluble nitrogen increased from 8% to 45% during Natto preparation, But the soluble protein nitrogen showed a slight increase. 2. Fractionation of the peptides using Dowex-50 resins showed that they consisted mostly of lower molecular weight peptides which increased in accordance with the progress of fermentation, especially after 30-hour period. 3. Sixteen known and two unknown amino acids, and three peptides with different Rf values were identified during the Natto preparation. Their appearance showed some difference in that phenylalanine appeared after 10 hours, methionine, after 20 hours and proline, after 30 hours, respectively. The three peptides appeared at the different stage of fermentation.

• Korean Journal of Organic Agriculture
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• v.21 no.3
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• pp.437-450
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• 2013

Miscanthus sacchariflorus var. No. 1 has been newly developed in Korea. This study was conducted to assess the feed value of M. sacchariflorus var. No. 1 at different growth and harvesting time. Total 3 different miscanthus - 1y4m (first shoot and harvested at 4 month), 2y4m (second shoot and harvested at 4 month) and 2y8m (second shoot and harvested at 8 month). Two experiments were carried out, In vitro rumen simulated fermentation and In situ dry matter digestibility (DMD). Ruminal pH at in vitro fermentation were higher in M. sacchariflorus var. No. 1 treatments compared to the rice straw (RS). In volatile fatty acid production, 1y4m resulted in higher acetate production than the other M. sacchariflorus var. No. 1 at higher maturity stages. Significant differences among treatments were observed in propionate and total volatile fatty acid (VFA) productions at 9, 24 and 48 h of incubation times. Higher ammonia nitrogen productions were found as increased maturity of M. sacchariflorus var. No. 1. At In situ experiment, high DMD was detected in the order of RS (60.51%) > 1y4m (57.65%) > 2y4m (57.63%) > 2y8m (46.28%). The results from this study indicate that young and early harvested M. sacchariflorus var. No. 1 are able to improve its nutrient values in the ruminant animal.

• Journal of agriculture & life science
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• v.51 no.5
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• pp.91-101
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• 2017

The in vitro experiment was conducted to ensure the supplemental level of spent Flammulina velutipes mushroom substrates(SMS) as an energy source in manufacturing of rye silage. Rye harvested at heading stage was ensiled with spent mushroom substrates of 0%(Control), 20%(R-20), 40%(R-40) and 60%(R-60) as fresh matter basis for 6week. The rumen fluid for preparation of in vitro solution was collected from two cannulated Holstein bulls fed a 40:60 concentrate:timothy diet. The experiment was conducted by 3, 6, 9, 12, 24, and 48 hrs of ncubation time with 3 replications. The silages were evaluated fermentation characteristics and dry matter digestibility(DMD) in vitro. The pH of in vitro solution was inclined to decrease with elapsing the incubation time, and that of the R-60 was significantly(p<0.05) lower than the other treatment at 48 hr of incubation. The microbial growth in vitro was inclined to increase with elapsing the incubation time, and that of the R-20 was significantly(p<0.05) greater than the Control at 48 hr of incubation. Gas production was greater(p<0.05) in the Control than the other treatments at 48 hr of incubation. In vitro dry matter digestibility(IVDMD) was higher with increasing the supplemental level of SMS, and was significantly(p<0.05) lower in the Control compared with other treatments throughout whole incubation time. The IVDMD for R-60 was the highest(p<0.05) among treatments at 24 hr and 48 hr of incubation. Considering of above results and the availability of SMS, SMS could be supplemented by 60% in fresh matter basis for rye silage fermentation.

• Food Science of Animal Resources
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• v.34 no.1
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• pp.40-48
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• 2014

In this study, the effects of two different commercial starter culture mixes and processing methodologies (traditional and heat process) on the lipolytic changes of fermented sausages manufactured with turkey meat were evaluated during processing stages and storage. Free fatty acid (FFA) value increased with fermentation and during storage over 120 d in all fermented sausage groups produced with both processing methodologies (p<0.05). After drying stage, free fatty acid values of traditional style and heat processed fermented sausages were between 10.54-13.01% and 6.56-8.49%, respectively. Thiobarbituric acid (TBA) values of traditionally processed fermented sausages were between $0.220-0.450mg{\cdot}kg^{-1}$, and TBA values of heat processed fermented sausages were in a range of $0.405-0.795mg{\cdot}kg^{-1}$. Oleic and linoleic acids were predominant fatty acids in all fermented sausages. It was seen that fermented sausage groups produced with starter culture had lower TBA and FFA values in comparison with the control groups, and heat application inhibited the lipase enzyme activity and had an improving effect on lipid oxidation. As a result of these effects, heat processed fermented sausages had lower FFA and higher TBA values than the traditionally processed groups.

• Journal of Microbiology
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• v.38 no.3
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• pp.160-168
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• 2000

A-factor I a microbial hormone that can positively control cell differentiation leading to spore formation and secondary metabolite formation in Streptomyces griseus. to identify a protease that is deeply involved in the morphological and physiological differentiation of Streptomyces, the proteases produced by Streptomyces griseus IFO 13350 and its A-factor deficient mutant strain, Streptomyces griseus HH1, as well as Streptomyces griseus HH1 transformed with the afsA gene were sturdied. In general Streptomyces griseus showed a higher degree of cell growth and protease activity in proportion to its ability to produce a higher amount of A-factor. In particular, the specific activity of the trypsin of Streptomyces griseus IFO 13350 was greatly enhanced more than twice compared with that of Streptomyces griseus HH1 in the later stage of growth. The specific activity of the metalloprotease of Streptomyces griseus HH1 was greatly enhanced more than twice compared with that of Streptomyces griseus IFO 13350, and this observation was reversed in the presence of thiostreptione, However, Streptomyces griseus HH1 transformed with the afsA gene showed a significantly decreased level of trypsin and metalloprotease activity compared with that of the HH1 strain. There was no significant difference between Streptomyces griseus IFO 13350 and HH1 strain in their chymotrypsin and thiol protease activity, yet the level of leu-amionpeptidase activity was 2 times higher in Streptomyces griseus HH1 than in strain IFO 13350 . Streptomyces griseus HH1 harboring afsA showed a similar level of enzyme activity , however, all the three protease activities sharply increased and the thiol protease activity was critically increased at the end of the fermentation. When a serine protease inhibitor, pefabloc SC, and metalloprotease inhibitor, EDTA, were applied to strain IFO 13350 to examine the in vivo effects of the protease inhibitors on the morpholofical differentiation, the formation of aerial meycelium and spores was delayed by two or three days.