• 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 Soil Science and Fertilizer
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• v.5 no.2
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• pp.1-38
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• 1972

This study, designed to establish a classification system of paddy soils and suitability groups on productivity and management of paddy land based on soil characteristics, has been made for the paddy soils on the Gimje-Mangyeong plains. The morphological, physical and chemical properties of the 15 paddy soil series found on these plains are briefly as follows: Ten soil series (Baeggu, Bongnam, Buyong, Gimje, Gongdeog, Honam, Jeonbug, Jisan, Mangyeong and Suam) have a B horizon (cambic B), two soil series (Geugrag and Hwadong) have a Bt horizon (argillic B), and three soil series (Gwanghwal, Hwagye and Sindab) have no B or Bt horizons. Uniquely, both the Bongnam and Gongdeog series contain a muck layer in the lower part of subsoil. Four soil series (Baeggu, Gongdeog, Gwanghwal and Sindab) generally are bluish gray and dark gray, and eight soil series (Bongnam, Buyong, Gimje, Honam, Jeonbug, Jisan, Mangyeong and Suam) are either gray or grayish brown. Three soil series (Geugrag, Hwadong and Hwagye), however, are partially gleyed in the surface and subsurface, but have a yellowish brown to brown subsoil or substrata. Seven soil series (Bongnam, Buyong, Geugrag, Gimje, Gongdeog, Honam and Hwadong) are of fine clayey texture, three soil series (Baeggu, Jeonbug and Jisan) belong to fine loamy and fine silty, three soil series (Gwanghwal, Mangyeong and Suam) to coarse loamy and coarse silty, and two soil series (Hwagye and Sindab) to sandy and sandy skeletal texture classes. The carbon content of the surface soil ranges from 0.29 to 2.18 percent, mostly 1.0 to 2.0 percent. The total nitrogen content of the surface soil ranges from 0.03 to 0.25 percent, showing a tendency to decrease irregularly with depth. The C/N ratio in the surface soil ranges from 4.6 to 15.5, dominantly from 8 to 10. The C/N ratio in the subsoil and substrata, however, has a wide range from 3.0 to 20.25. The soil reaction ranges from 4.5 to 8.0. All soil series except the Gwanghwal and Mangyeong series belong to the acid reaction class. The cation exchange cpacity in the surface soil ranges from 5 to 13 milliequivalents per 100 grams of soil, and in all the subsoil and substrata except those of a sandy texture, from 10 to 20 milliequivalents per 100 grams of soil. The base saturation of the soil series except Baeggu and Gongdeog is more than 60 percent. The active iron content of the surface soil ranges from 0.45 to 1.81 ppm, easily-reduceable manganese from 15 to 148 ppm, and available silica from 36 to 366 ppm. The iron and manganese are generally accumulated in a similar position (10 to 70cm. depth), and silica occurs in the same horizon with that of iron and manganese, or in the deeper horizons in the soil profile. The properties of each soil series extending from the sea shore towards the continental plains change with distance and they are related with distance (x) as follows: y(surface soil, clay content) = $$-0.2491x^2+6.0388x-1.1251$$ y(subsoil or subsurface soil, clay content) = $$-0.31646x^2+7.84818x-2.50008$$ y(surface soil, organic carbon content) = $$-0.0089x^2+0.2192x+0.1366$$ y(subsoil or subsurface soil, pH) = $$-0.0178x^2-0.04534x+8.3531$$ Soil profile development, soil color, depositional and organic layers, soil texture and soil reaction etc. are thought to be the major items that should be considered in a paddy soil classification. It was found that most of the soils belonging to the moderately well, somewhat poorly and poorly drained fine and medium textured soils and moderately deep fine textured soils over coarse materials, produce higher paddy yields in excess of 3,750 kg/ha. and most of the soils belonging to the coarse textured soils, well drained fine textured soils, moderately deep medium textured soils over coarse materials and saline soils, produce yields less than 3,750kg/ha. Soil texture of the profile, available soil depth, salinity and gleying of the surface and subsurface soils etc. seem to be the major factors determining rice yields, and these factors are considered when establishing suitability groups for paddy land. The great group, group, subgroup, family and series are proposed for the classification categories of paddy soils. The soil series is the basic category of the classification. The argillic horizon (Bt horizon) and cambic horizon (B horizon) are proposed as two diagnostic horizons of great group level for the determination of the morphological properties of soils in the classification. The specific soil characteristics considered in the group and subgroup levels are soil color of the profile (bluish gray, gray or yellowish brown), salinity (salic), depositonal (fluvic) and muck layers (mucky), and gleying of surface and subsurface soils (gleyic). The family levels are classified on the basis of soil reaction, soil texture and gravel content of the profile. The definitions are given on each classification category, diagnostic horizons and specific soil characteristics respectively. The soils on these plains are classified in eight subgroups and examined under the existing classification system. Further, the suitability group, can be divided into two major categories, suitability class and subclass. The soils within a suitability class are similar in potential productivity and limitation on use and management. Class 1 through 4 are distinguished from each other by combination of soil characteristics. Subclasses are divided from classes that have the same kind of dominant limitations such as slope(e), wettness(w), sandy(s), gravels(g), salinity(t) and non-gleying of the surface and subsurface soils(n). The above suitability classes and subclasses are examined, and the definitions are given. Seven subclasses are found on these plains for paddy soils. The classification and suitability group of 15 paddy soil series on the Gimje-Mangyeong plains may now be tabulated as follows.