• Title/Summary/Keyword: microbial enzyme

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Effects of Dietary Supplementation of Coffee Meal on Intestinal Enzyme Activity, Biochemical Profiles and Microbial Population in Broiler Chicks (커피박 첨가가 육계의 소장 효소 활성도, 생화학 지표 및 장내 미생물 균총에 미치는 영향)

  • Ko, Young-Hyun;Yun, Seo-Hyun;Song, Min-Hae;Kim, Se-Yun;Kim, Jong-Sun;Kim, Hyoun-Wook;Jang, In-Surk
    • Korean Journal of Poultry Science
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    • v.41 no.2
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    • pp.105-113
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    • 2014
  • The current study was performed to investigate the effects of dietary supplementation of dried coffee meal (CM) on growth performance, intestinal and blood biochemical index, intestinal enzymes, and cecal microbial populations. A total of 162, 3-day-old male broiler chicks were randomly allocated into three dietary groups: control group (CON), basal diet added with 0.5% CM (CM I), and basal diet added with 1.0% CM (CM II). Dietary supplementation of CM did not change bird performance and the relative weight of intestinal mucosal tissues. The birds fed the diet supplemented with CM (0.5 and 1.0%) significantly decreased mucosal glucose concentration (P<0.05) without affecting blood glucose level compared with those fed control diet. The level of blood aspartate aminotransferase (AST) significantly increased in CM II group (P<0.05) without affecting ${\gamma}$-glutamyl transpeptidase (${\gamma}$-GTP) compared with that in the CON group. The specific activity of intestinal maltase, leucine aminopeptidase (LAP) and alkaline phosphatase (ALP) were not affected by dietary supplementation of CM, whereas sucrase activity in birds fed the diet supplemented with CM was decreased (P<0.05) compared to that in the control birds. The colony forming units (CFU) of E. coli in the cecum of CM-fed birds was significantly decreased (P<0.05) compared with that of control birds without changing the CFU of Lactobacillus. In conclusion, dietary supplementation of lower level of CM (0.5%) can be used as a beneficial feed resource without liver toxicity in broiler chicks.

Effect of Urushiol-Free Extracts from Fermented-Rhus verniciflua Stem Bark with Fomitella fraxinea on the Fermentation Characteristics of Doenjang (Soybean Paste) (장수버섯 배양법에 의해 urushiol이 제거된 발효옻 추출물이 된장발효에 미치는 영향)

  • Choi, Han-Seok;Jeong, Seok-Tae;Choi, Ji-Ho;Kang, Ji-Eun;Kim, Eugene;Noh, Jong-Min;Kim, Myung-Kon
    • The Korean Journal of Mycology
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    • v.40 no.4
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    • pp.244-253
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    • 2012
  • The effect of fermented Rhus verniciflua stem bark (FRVSB) extract on the microbial count, enzyme activity, concentrations of free amino acids and organic acids, and physiochemical properties of doenjang (soybean paste) was evaluated during brine fermentation. The FRVSB extract increased the total free amino acid concentration by 1.3-3.1-fold on the $42^{nd}$ day of brine fermentation. After the filtration of brine, the following microbial counts were obtained in the doenjang: bacteria, $0.3{\times}10^8-12.0{\times}10^8$ cfu/g; mold, $3.0{\times}10^4-21.0{\times}10^4$ cfu/g; yeast, $1.0{\times}10^4-2.0{\times}10^4$ cfu/g; Escherichia coli, not detected; and Bacillus cereus, $3.0{\times}10^2-25.0{\times}10^2$ cfu/g. The FRVSB extract addition enhanced the protein and starch degrading activity by 13.8-26.0% and 16.1-35.1%, respectively. The extract increased the total free amino acid content by 1.4-3.0-fold. Lactic acid, acetic acid, and pyroglutamic acid were the predominant organic acids in doenjang. Moreover, the proximate composition, pH, moisture, ash, salt, and amino nitrogen content were increased.

Development of a Molecular Selection Marker for Bacillus licheniformis K12 (Bacillus licheniformis K12 균주 분자 선발 마커 개발)

  • Young Jin Kim;Sam Woong Kim;Tae Wok Lee;Won-Jae Chi;Woo Young Bang;Ki Hwan Moon;Tae Wan Kim;Kyu Ho Bang;Sang Wan Gal
    • Journal of Life Science
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    • v.33 no.10
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    • pp.808-819
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    • 2023
  • This study was conducted to develop a selection marker for the identification of the Bacillus licheniformis K12 strain in microbial communities. The strain not only demonstrates good growth at moderate temperatures but also contains enzymes that catalyze the decomposition of various polymer materials, such as proteases, amylases, cellulases, lipases, and xylanases. To identify molecular markers appropriate for use in a microbial community, a search was conducted to identify variable gene regions that show considerable genetic mutations, such as recombinase, integration, and transposase sites, as well as phase-related genes. As a result, five areas were identified that have potential as selection markers. The candidate markers were two recombinase sites (BLK1 and BLK2), two integration sites (BLK3 and BLK4), and one phase-related site (BLK5). A PCR analysis performed with different Bacillus species (e.g., B. licheniformis, Bacillus velezensis, Bacillus subtilis, and Bacillus cereus) confirmed that PCR products appeared at specific locations in B. licheniformis: BLK1 in recombinase, BLK2 in recombinase family protein, and BLK3 and BLK4 as site-specific integrations. In addition, BLK1 and BLK3 were identified as good candidate markers via a PCR analysis performed on subspecies of standard B. licheniformis strains. Therefore, the findings suggest that BLK1 can be used as a selection marker for B. licheniformis species and subspecies in the microbiome.

Studies on the Enzyme from Arthrobacter luteus Accelerating the Lysis of Yeast Cell Walls -II. Separation of the Factor Accelerating the Lysis of Yeast Cell Walls from the Preparation of Crude Zymolyase and Partial Purification of the Zymolyase with the Sephadex G-75 Gel- (Arthrobacter luteus가 생산(生産)하는 효모세포벽(酵母細胞壁) 용해촉진효소(溶解促進酵素)에 관(關)한 연구(硏究) -제 2 보(第2報) : Crude Zymolyase 표품중(標品中)으로부터 효모(酵母) 세포벽(細胞壁) 용해(溶解) 촉진(促進) 인자(因子)의 분리(分離) 및 Sephadex G-75 Gel에 의한 Zymolyase의 부분(部分) 정제(精製)-)

  • Oh, Hong-Rock;Shimoda, Tadahisa;Funatsu, Masaru
    • Korean Journal of Food Science and Technology
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    • v.12 no.4
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    • pp.254-262
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    • 1980
  • A series of experiment were carried out to separate the factor accelerating the lysis of cell wall of $Saccharomyces\;sak{\acute{e}}$ from the preparation of crude zymolyase obtained from Arthrobacter luteus. An attempt was also made to purify the enzyme which is essential for the study on the separation of the factor. The results are summarized as follows: 1. Crude zymolyase was fractionated 5 peaks $(A{\sim}E)$ containing three peaks $(A{\sim}C)$ passed through the column by the chromatography on Biogel CM-30. 2. Among the five peaks, peak E (protease fraction) was found to contain the factor accelerating the lytic activity of the zymolyase. 3. L-c fraction purified in almost free form from the nonlytic ${\beta}-1$, 3-glucanase, protease and inert protein by the affinity adsorption chromatography with Sephadex G-75 gel was obtained from zymolyase fraction (peak D). When it was subjected to polyacrylamide gel disc electrophoresis, only one clear protein band was observed at pH 4. 5, but still detected two or more band at pH 8. 3.

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Microbiological and Enzymological Studies on the Flavor Components of Sea Food Pickles (젓갈등속(等屬)의 정미성분(呈味成分)에 관(關)한 미생물학적(微生物學的) 및 효소학적(酵素學的) 연구(硏究))

  • Lee, Ke-Ho
    • Applied Biological Chemistry
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    • v.11
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    • pp.1-27
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    • 1969
  • More than thirty kinds of sea food pickles have been eaten in Korea. Out of these salted yellow tail pickle, salted clam pickle, salted oyster pickle, and salted cuttlefish pickle were employed for the analysis of their components, identification of main fermenting microbes, and determination of enzyme characteristics concerned. Also studied was the effect of enzymic action of microbes, which are concerned with the fermenting of pickles, on the production of flavorous 5'-mononucleotides and amino acids. The results are summarized as follows: 1. Microflora observed in the pickles are: (a) Total count of viable cells after 1-2 months of pickling was found to be $10^7$ and that after 6 months decreased to $10^4$. (b) Microbial occurence in the early stage of pickling was observed to be 10-20% Micrococcus spp., 10-20% Brevibacterium spp., 0-30% Sarcina spp., 20-30% Leuconostoc spp., ca 30% Bacillus spp., 0-10% Pseudomonas spp., 0-10% Flavobacterium spp., and 0-20% yeast. (c) Following the early stage of pickling, mainly halophilic bacteria such as Bacillus subtilis, Leuconostoc mesenteroides, Pediococcus halophilus and Sarcina litoralis, were found to exhibit an effect on the fermentation of pickle and their enzyme activities were in direct concern in fermentation of pickles. (d) Among the bacteria participating in the fermentation, Sarcina litoralis 8-14 and 8-16 strains were in need of high nutritional requirement and the former was grown only in the presence of purine, pyrimidine and cystine and the latter purine, pyrimidine and glutamic acid. 2. Enzyme characteristics studied in relation to the raw materials and the concerned microbes isolated are as follows: (a) A small amount of protease was found in the raw materials and 30-60% decrease in protease activity was demonstrated at 7% salt concentration. (b) Protease activity of halophilic bacteria, Bacillus subtilis 7-6, 11-1, 3-6 and 9-4 strains, in the complete media decreased by 10-30% at the 7% salt concentration and that of Sarcina litoralis 8-14 and 8-16 strains decreased by 10-20%. (c) Proteins in the raw materials were found to be hydrolyzed to yield free amino acids by protease in the fermenting microbes. (d) No accumulation of flavorous 5'-mononucleotides was demonstrated because RNA-depolymerase in the raw materials and the pickles tended to decompose RNA into nucleoside and phosphoric acid. (e) The enzyme produced in Bacillus subtilis 3-6 strain isolated from the salted clam pickles, was ascertained to be 5'-phosphodiesterase because of its ability to decompose RNA and thus accumulating 5'-mononucleotide. (f) It was demonstrated that the activity of phosphodiesterase in Bacillus subtilis 3-6 strain was enhanced by some components in the corn steep liquor and salted clam pickle. The enzyme activity was found to decrease by 10-30% and 40-60% at the salt concentration of 10% and 20%, respectively. 3. Quantitative data for free amino acids in the pickles are as follows: (a) Amounts of acidic amino acids such as glutamic and aspartic acids in salted clam pickle, were observed to be 2-10 times other pickles and it is considered that the abundance in these amino acids may contribute significantly to the specific flavor of this food. (b) Large amounts of basic amino acids such as arginine and histidine were found to occur in salted yellow tail pickle. (c) It is much interesting that in the salted cuttlefish pickle the contents of sulfur-containing amino acids were exceedingly high compared with those of others: cystine was found to be 17-130 times and methionine, 7-19 times. (d) In the salted oyster pickle a high content of some essential amino acids such as lysine, threonine, isoleucine and leucine, was demonstrated and a specific flavor of the pickle was ascribed to the sweet amino acids. Contents of alanine and glycine in the salted oyster pickle were 4 and 3-14 times as much as those of the others respectively. 4. Analytical data for 5'-mononucleotides in the pickles are as follows: (a) 5'-Adenylic acid and 3'-adenylic acid were found in large amounts in the salted yellow tail pickle and 5'-inosinic acid in lesser amount. (b) 5'-Adenylic acid, especially 3'-adenylic acid predominated in amount in the salted oyster pickle over that in the other pickles. (c) The salted cuttlefish pickle was found to contain only 5'-adenylic acid and 3'-adenylic acid. It has become evident from the above fact that clam and the invertebrate lack of adenylic deaminase and contain high content of adenylic acid. Thus, they were demonstrated to be the AMP-type. (d) 5'-Inosinic acid was contained in the salted yellow tail pickle in a significant concentration, and it might be considered to be IMP-type. 5. Comparative data for flavor with regard to the flavorous amino acids and the contents of 5'-mononucleotides are: (a) A specific flavor of salted yellow tail pickle was ascribed to the abundance in glutamic acid and aspartic acid, and to the existence of a small amount of flavorous 5'-inosinic acid. The combined effect of these components was belived to exhibit a synergistic action in producing a specific fiavor to the pickle. (b) A specific flavor of salted clam pickle has been demonstrated to be attributable to the richness in glutamic acid and aspartic acid rather than to that of 5'-mononucleotides.

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Screening of Brewing Yeasts and Saccharifying Molds for Foxtail Millet-Wine Making (제주민속 좁쌀약주 발효를 위한 우수균주의 선발)

  • Kim, Ji-Yong;Koh, Jeong-Sam
    • Applied Biological Chemistry
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    • v.47 no.1
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    • pp.78-84
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    • 2004
  • In order Nuruk to improve the quality of millet wine, a traditional Jeju cereal wine, yeasts and molds were isolated from 35 kinds of Nuruk collected nationwide. Isolated strains were screened for saccharification of starch and brewing of millet wine. Fermentation characteristics of millet wine with different types of Nuruk were also investigated. The average number of microbial populations in the Nuruk were $6.4{\times}10^5{\sim}4.5{\times}10^7\;cfu/g$ for molds and $1.4{\times}10^4{\sim}7.7{\times}10^7\;cfu/g$ for yeasts. Among the 169 strains of molds and 103 strains of yeasts, 16 strains were screened for saccharifying activity on starch as a substrate, and one yeast strain was screened for the brewing of millet wine. A8-3, supposed as Aspergillus sp., showed the highest enzyme activities of glucdamylase, ${\alpha}-amylase$ and xylanase while B23-3 strain, supposed as Rhizopus sp., showed the highest saccharifying activity. A10-4, supposed as Saccharomyces sp., showed the highest level of weight loss from $CO_2$ evolution, sugar and alcohol tolerance during fermentation. When the Nuruk was made after inoculation with the selected strains, saccharifying activity was higher for the co-cultivation of A8-3 and B23-3 than individual cultivation of each strain. Similar saccharifying activities were shown in both disc-type and pellet-type Nuruk. It was suggested that pellet-type Nuruk could improve fermentation yield. The collected Nuruk consisted of $10{\sim}13%$ moisture, $55{\sim}70%$ total sugar, $10{\sim}18%$ crude protein, $0.2{\sim}1.0%$ crude fat and $1.8{\sim}2.1%$ ash. The Nuruk made in this study was composed of $12{\sim}15%$ moisture, $61{\sim}71%$ total sugar, $15{\sim}20%$ crude protein, $0.4{\sim}1.5%$ crude fat and $1.1{\sim}1.5%$ ash.

Modulation of Cellulalr Quinone Reductase Inducibility by Roasting Treatment and Acid Hydrolysis of Perilla (들깨의 볶음처리와 산가수분해에 의한 세포모델계 Quinone Reductase 활성유도능의 변화)

  • Hong, Eun-Young;Kang, Hee-Jung;Kwon, Chong-Suk;Nam, Young-Jung;Suh, Myung-Ja;Kim, Jong-Sang
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.2
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    • pp.186-192
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    • 1997
  • Increased activities of phase 2 enzymes including quinone reductase(QR) have been reported to be associated with protection of animals from neoplastic, mutagenic, and other toxic effects of many carcinogens. In previous study, we found that methanol extract of roasted and defatted perilla meal induced the activity of quinone reductase, an anticarcinogenic marker enzyme, in murine hepalc1c7 cells. Current study showed that unroasted perilla had a limited QR-inducing activity, suggesting that roasting cause the generation of active component(s). Thus we hypothesized that QR inducer in perilla might be covalently linked to sugar moiety and released during roasting process. Methanol extract of defatted raw perilla was subject to acid treatment in order to hydrolyze the potential sugar moiety. Prolonged hydrolysis of methanol extract of defatted raw perilla at $98{\sim}100^{\circ}C$ increased the ability to induce cytosolic QR activity of hepalclc7 cells. Furthermore roasting at 180 and $200^{\circ}C$ resulted in significant induction of QR activity. The result strongly support the idea that QR inducer(s) is present in bound form in raw perilla and released during roasting. Cellular QR activity was induced proportionately with the increase of concentration of methanol extract of roasted perilla. The induction of QR by defatted perilla was also examined in the cytosols of liver, small intestine, stomach, lung and kidney of male ICR mice. Induction patterns showed specificity with respect to target tissue and roasting of perilla. Unroasted perilla meal (defatted) significantly induced QR in liver and lung, while roasted perilla meal induced QR in liver and stomach. The observation that raw perilla showed similar QR induction patterns to roasted perilla is consistent with our proposal that QR inducer(s) is present in bound form and released by physical and chemical treatments as digestive or microbial enzymes could release the inducers from inactive glycoside forms in gastrointestinal tract of mice. In conclusion, perilla could exert protective effect against chemically induced carcinogenesis by inducing phase 2 enzymes in biological systems regardless of chemical and physical process such as roasting.

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Effect of Phosphate Bio fertilizer Produced by Enterobacter intermedium on Rhizosphere Soil Properties and Lettuce Growth (Enterobacter intermedium으로 제조된 인산생물비료가 토양 특성 및 상추의 생육에 미치는 영향)

  • Park, Bum-Ki;Na, Jung-Heang;Hwang-Bo, Hoon;Lee, In-Jung;Kim, Kil-Yong;Kim, Yong-Woong
    • Korean Journal of Soil Science and Fertilizer
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    • v.38 no.1
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    • pp.15-24
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    • 2005
  • Enterobacter intermedium oxidizes glucose to gluconic acid and sequentially converts gluconic acid to 2-ketogluconic acid (2-KGA) under aerobic condition. Shaking incubation of E. intermedium in a broth medium containing 22.5 g glucose, 8.2 g gluconic acid and 40 g rock phosphate per liter resulted in $1028mg\;L^{-1}$ soluble phosphate. The culture broth was used as phosphate bio-fertilizer (PBF) in this experiment. To evaluate PBF produced by E. intermedium on lettuce growth, five treatments (PBF1/3, PBF2/3, PBF3/3, BP, and MF) were used. In MF and BP treatments, $P_2O_5$ 5.9 kg of mineral fertilizer per 10a was added, while in PBF1/3, PBF2/3, and PBF3/3 treatments, culture broth containing one third, two third, and same amount of soluble $P_2O_5$ 5.9 kg per 10a was applied, respectively. At 20, 35, and 50 days after transplanting of lettuce, plant growth components, biomass, enzyme activities and soil chemical properties were analyzed. Dehydrogenase activity and available phosphate concentration of rhizosphere in phosphate bio fertilizer treatments (PBF1/3, PBF2/3, PBF3/3) were generally higher compared to MF and BP treatments. Soil biomass in PBF3/3 treatment was significantly higher than MF and BP treatments at early growth stage. However, there was no significant difference among all treatments with time. Plant fresh weights in PBF1/3, PBF2/3, and MF treatments were better than those in BP and PBF3/3 treatments. However, in PBF2/3 treatment the highest fresh weight was discovered where alkaline phosphatase activity was generally higher than other treatments at 35 and 50 days. Enhancement of lettuce growth at 35 and 50 days in PBF2/3 treatment was associated with greater phosphate uptake in lettuce tissue. As regarding all results, PBF showed better lettuce growth compared to mineral phosphate fertilizer where PBF2/3 treatment resulted in increase of lettuce fresh weight by 23% and phosphate uptake by 50%.

Application of Earthworm Casting-derived Biofilter Media for Hydrogen Sulfide Removal (지렁이 분변토를 이용한 생물담체가 충전된 바이오필터에 의한 황화수소 제거)

  • Yoo, Sun-Kyoung;Lee, Eun-Young
    • Journal of Korean Society of Environmental Engineers
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    • v.29 no.7
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    • pp.820-825
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    • 2007
  • Earthworm casting was the natural fertilizer that contained high concentrations of nutrients such as nitrogen, phosphate and potassium and of over $10^8$ CFU/ml of microorganisms. Greater than 80% of feed was excreted through the fermentation by the intestinal enzyme, after worm had eaten feeds such as fallen leaves and rotten roots under the ground. Also, the soil structure of casting was known to be very efficient in the aspects of the porosity, the water permeability, and deodorizing activities. In this research, the biofilter packed with a biomedia made of casting and waste polyurethane foam, a binder, which helped to improve the durability and perpetuity of casting, was investigated to degrade malodorous hydrogen sulfide gas. The biomedia had no need of extra supply of nutrients and of microbial inoculations. On the beginning of the operations, it showed 100% removal of hydrogen sulfide gas without lag phase. At SV of 50 $h^{-1}$, hydrogen sulfide gas from the outlet of the biofilter was not detected, when inlet concentration increased to 450 ppmv. After that, removal efficiency decreased as increasing inlet hydrogen sulfide concentration. Hydrogen sulfide removal was maintained at almost 93% until inlet concentration was increased up to 950 ppmv, at which the elimination capacity of $H_2S$ was 61.2 g $S{\cdot}m^{-3}{\cdot}h^{-1}$. Maximum elimination capacity guaranteing 90% removal was 61.2, 65.9, 84.7, 89.4 g $S{\cdot}m^{-3}{\cdot}h^{-1}$ at SV ranging from 50 $h^{-1}$ to 300 $h^{-1}$, but was 59.3 g $S{\cdot}m^{-3}{\cdot}h^{-1}$ at SV of 400 $h^{-1}$. The results calculated from Michaelis-Menten equation revealed that $V_m$ increased from 66.04, 88.96, 117.35, 224.15, to 227.54 g $S{\cdot}m^{-3}{\cdot}h^{-1}$ with increasing space velocity in the range of 50 $h^{-1}$ to 400 $h^{-1}$. However, saturation constant$(K_s)$ decreased from 79.97 ppmv to 64.95 and 65.37 ppmv, and then increased to 127.72 and 157.43 ppmv.

Comparative Assessment of Specific Genes of Bacteria and Enzyme over Water Quality Parameters by Quantitative PCR in Uncontrolled Landfill (정량 PCR을 이용한 비위생 매립지의 특정 세균 및 효소 유전자와 수질인자와의 상관관계 평가)

  • Han, Ji-Sun;Sung, Eun-Hae;Park, Hun-Ju;Kim, Chang-Gyun
    • Journal of Korean Society of Environmental Engineers
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    • v.29 no.8
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    • pp.895-903
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    • 2007
  • As for the increasing demanding on the development of direct-ecological landfill monitoring methods, it is needed for critically defining the condition of landfills and their influence on the environment, quantifying the amount of enzymes and bacteria mainly concerned with biochemical reaction in the landfills. This study was thus conducted to understand the fates of contaminants in association with groundwater quality parameters. For the study, groundwater was seasonally sampled from four closed unsanitary landfills(i.e. Cheonan(C), Wonju(W), Nonsan(N), Pyeongtaek(P) sites) in which microbial diversity was simultaneously obtained by 16S rDNA methods. Subsequently, a number of primer sets were prepared for quantifying the specific gene of representative bacteria and the gene of encoding enzymes dominantly found in the landfills. The relationship between water quality parameters and gene quantification were compared based on correlation factors. Correlation between DSR(Sulfate reduction bacteria) gene and BOD(Biochemical Oxygen Demand) was greater than 0.8 while NSR(Nitrification bacteria-Nitrospira sp.) gene and nitrate were related more than 0.9. A stabilization indicator(BOD/COD) and MTOT(Methane Oxidation bacteria), MCR(Methyl coenzyme M reductase), Dde(Dechloromonas denitrificans) genes were correlated over 0.8, but ferric iron and Fli(Ferribacterium limineticm) gene were at the lowest of 0.7. For MTOT, it was at the highest related at 100% over BOD/COD. In addition, anaerobic genes(i.e., nirS-Nitrite reductase, MCR. Dde, DSR) and DO were also related more than 0.8, which showing anaerobic reactions generally dependant upon DO. As demonstrated in the study, molecular biological investigation and water quality parameters are highly co-linked, so that quantitative real-time PCR could be cooperatively used for assessing landfill stabilization in association with the conventional monitoring parameters.