• Title/Summary/Keyword: 기질 분해율

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Purification and Characterization of Chitinase from Antagonistic Bacteria Pseudomonas sp. 3098. (생물방제균 Pseudomonas sp. 3098이 생산하는 Chitinase의 정제 및 특성)

  • 이종태;김동환;도재호;김상달
    • Microbiology and Biotechnology Letters
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    • v.26 no.6
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    • pp.515-522
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    • 1998
  • Plant root rotting fungi, Fusarium solani are suppressed their growth by the chitinase which is produced from the antagonistic soil bacteria. The chitinase producable antagonistic bacterium Pseudomonas sp. 3098 was selected as a powerful biocontrol agent of F. solani from ginseng rhizosphere. The antagonistic Pseudomonas sp. 3098 was able to produce a large amount of extracellular chitinase which is key enzyme in the decomposition of fusarial hypal walls. The chitinase was purified from cultural filtrate of Pseudomonas sp. 3098 by the procedure of ammonium sulfate precipitation, anion exchange chromatography, gel filtration on Bio-Gel P-100, and 1st and 2nd hydroxyapatite chromatography. The molecular mass of the purified enzyme was ca. 45 kDa on SDS-FAGE. The optimal pH and temperature for the activity of purified chitinase were 5.0 and 45$^{\circ}C$, respectively. The enzyme was stable in pH range of 5.0 to 9.0 up to 5$0^{\circ}C$ The enzyme was significantly inhibited by metal compounds such as FeCl$_2$, AgNO$_3$ and HgCl$_2$, and was slightly inhibited by p-CMB, iodoacetic acid, urea, 2,4-DNP and EDTA. The enzyme had ability of digestion on colloidal chitin and chitin from shrimp shell, but could not digest chitosan and chitin from crab shell. Km value of the enzyme was 0.11% on colloidal chitin, and the maximum hydrolysis rate of the enzyme was 34% on colloidal chitin.

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Conditions of Hemoglobin Hydrolysis and Separation for the Production of Enriched Heme-iron (고농도 Heme-iron의 생산을 위한 Hemoglobin의 가수분해 및 분리 조건)

  • Kang, In-Kyu;In, Man-Jin;Oh, Nam-Soon
    • Applied Biological Chemistry
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    • v.44 no.4
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    • pp.219-223
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    • 2001
  • Effects of hemoglobin (Hgb) concentration and degree of hydrolysis (DH) of Hgb on the separation of heme-iron were examined to produce highly enriched heme-iron from Hgb hydrolysate. Separation efficiency of Hgb hydrolysate with different DH was studied at wide pH range (pH $1.0{\sim}11.0$). Separation efficiency expressed as heme-iron/peptide ratio increased with decreasing Hgb concentration. When 5% Hgb (pH 10.0) was hydrolyzed using commercially available Esperase for 5 h at $50^{\circ}C$, DH was 25%. The precipitation of heme-iron-enriched peptides were remarkably high at pH range $3{\sim}6$. Optimal pH range for heme-iron with high heme-iron/peptide ratio shifted to acidic pH with increasing DHs of Hgb. The enriched heme-iron fraction in the precipitates showed a single band through urea-SDS-PAGE, with a molecular mass of 1 kDa. In the dry heme-iron product produced in a pilot bioreactor, content of heme-iron and heme-iron/peptide ratio were 27.1 and 38.7%, respectively, and production yield was 9.3%.

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Comparison of antioxidant capacity of protein hydrolysates from 4 different edible insects (식용곤충별 단백가수분해물의 항산화 활성 비교)

  • Jang, Hyun-Young;Park, Chae-Eun;Lee, Syng-Ook
    • Korean Journal of Food Science and Technology
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    • v.51 no.5
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    • pp.480-485
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    • 2019
  • The present study was conducted to compare antioxidant capacities of protein hydrolysates from four different edible insects (Protaetia brevitarsis larvae, Allomyrina dichotoma larvae, Gryllus bimaculatus imago, and Tenebrio molitor larvae) which have recently been registered as food varieties in Korea. Protein hydrolysates were prepared from each insect using enzymatic hydrolysis using alcalase, and were then separated into a fraction containing ${\leq}3kDa$. According to $RC_{50}$ values and trolox equivalent antioxidant capacity results obtained from five different antioxidant analyses, the Gryllus bimaculatus (GB) hydrolysate showed relatively high levels of antioxidant capacity and, in particular, the GB hydrolysate showed considerably strong antioxidant activities in 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging and in ferric reducing antioxidant power (FRAP) assays. The GB hydrolysate also showed the strongest inhibitory effect on peroxidation of linoleic acid, and its rate of inhibition at $100{\mu}g/mL$ on day 3 of treatment was 60.26%. These results suggest that protein hydrolysates from edible insects including GB represent potential sources of natural antioxidants.

Nutrient Dynamics in Decomposing Leaf Litter and Litter Production at the Long-Term Ecological Research Site in Mt. Gyebangsan (계방산 장기생태조사지의 낙엽 생산량 및 낙엽 분해에 따른 양분 동태)

  • Lee, Im-Kyun;Lim, Jong-Hwan;Kim, Choon-Sig;Kim, Young-Kul
    • Journal of Ecology and Environment
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    • v.29 no.6
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    • pp.585-591
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    • 2006
  • We measured the litterfall quantity and investigated the nutrient dynamics in decomposing litter for three years at the LTER sites installed in a deciduous broadleaf natural forest in Mt. Gyebangsan, South Korea. Litterfall production was significantly different among the sampling dates, whereas it was not significantly different among the years. The total annual mean litterfall production for three years was 6,593 kg $ha^{-1}$ $yr^{-1}$ and leaf litter accounted for 82.6% of the litterfall. The leaf litter quantity was highest in Quercus mongolia, followed by leaf of other species, Betula schmidtii, Kaplopanax pictus, Acer pseudo-sieboldianum, etc., which are dominant tree species in the site. The mass loss from the decomposition of leaf litter was fastest in Cortinus controversa (100%), followed by A. preudo-sieboldianum, K. pictus, and B. schmidtii. 100% of litter for C. controversa, 96.1% for A. pseudo-sieboldianum, 92.8% for K. pictus decomposed, while 66.2% of litter for Q. mongolia decayed for 1,003 days. The lower rate of the mass loss in the litter of Q. mongolia may be attributed to the difference in substrate quality, such as lower nutrient concentrations compared with those of other tree species. The concentrations of N, P, and Ca for five litter types increased over time, while the concentrations of K and Mg decreased over time. Compared with the nutrients in the litter of Q. mongolia, the nutrients (N, P, K, Ca, Mg) in the litter of other species, C. controversa, A. pseudo-sieboldianum, and K. pictus, were released more rapidly. The results showed that the mass loss and the nutrient dynamics in the litter are variable depending on the tree species even in the same site conditions.

Enzymatic Saccharification of Citrus Peel by Aspergillus sp. GF 015 (분리균 Aspergillus sp. GF015를 이용한 감귤과피(柑橘果皮)의 당화(糖化))

  • Park, Seok Kyu;Sung, Nack Kie;Chun, Hyo Kon
    • Microbiology and Biotechnology Letters
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    • v.13 no.1
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    • pp.59-64
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    • 1985
  • In order to utilize Citrus peel as fermentative substrate of microorganisms, enzymatic saccharification of Citrus peel by the crude enzyme of Aspergillus sp. GF 015 isolated and identified from nature was investigated. When the fungus was cultured at $27^{\circ}C$ for 3 days in wheat bran medium containing 0.6% $NH_4NO_3$ and 0.05% $KH_2PO_4$, the maximal production of the enzyme was observed. Optimal conditions for enzymatic reaction of crude enzyme were 15ml(97.5 unit)/g of enzyme solution to Citrus peel powder ratio, pH4.0, $45^{\circ}C$ of temperature and 12 hours of reaction time. As the result of saccharifying Citrus peel under optimum conditions, reducing sugar on the weight of dry matter was formed 60.2% and saccharifying rate was 76.3%. The sugar solution obtained were mainly composed of glucose, xylose and galacturonic acid. Hydrolyzing enzymes produced by Aspergillus sp. GF 015 were pectinase, cellulase and xylanase.

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Microbial Conversion of Woody Waste into Sugars and Feedstuff (II) - Production of Cellulolytic Enzymes from Aspergillus fumigatus and Saccharification of Popla Wood (미생물(微生物)에 의한 목질자원(木質資源)의 당화(糖化) 및 사료화(飼料化)에 관(關)한 연구(硏究) (II) - Aspergillus fumigatus KC-1으로부터 섬유소 분해 효소의 생산 및 현사시나무의 효소가수분해)

  • Chung, Ki-Chul;Huh, Jeong-Weon;Myung, Kyu-Ho;Kim, Yoon-Soo
    • Journal of the Korean Wood Science and Technology
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    • v.15 no.4
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    • pp.18-25
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    • 1987
  • The cellulolytic activities of Aspergillus fumigatus KC-1 was investigated, which showed the most active producer of cellulase among the 256 strains of cellulose-decomposing microorganisms screened in our laboratory. All the examined cellulolytic activities (filter paper-, Avicel-, cotton-, CMC-, salicin- and xylansaccharifying activity) in a culture of A. fumigatus KC-1 grown on 1% popular sawdust pretreated with peroxide alkaline reached a maximum within 4-5 days. The optimum pH and temperature for the enzymatic activity was found to be pH 4.5 and $60^{\circ}C$ respectively. The sawdust of poplar wood delignified with 1% NaOH and 20% peracetic acid succesively recorded the highest hydrolysis rate in the tests of enzymatic saccharification. The major end product of hydrolysis of poplar wood with the cellulolytic enzymes obtained from A. fumigatus KC-1 was glucose with small amount of cellobiose and xylose. It can be concluded from these results that A. fumigatus KC-1 is an advantagous source of a cellulase that is capable of hydrolyzing cellulose to glucose rapidly. The influence of degree of delignification, substrate size and its concentration on the rate of hydrolysis of poplar wood was also discussed.

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Thermal Acid Hydrolysis Pretreatment, Enzymatic Saccharification and Ethanol Fermentation from Red Seaweed, Gracilaria verrucosa (꼬시래기 홍조류로부터 열산가수분해, 효소당화 및 에탄올 발효)

  • Ra, Chae Hun;Choi, Jin Gyu;Kang, Chang-Han;Sunwoo, In Yung;Jeong, Gwi-Taek;Kim, Sung-Koo
    • Microbiology and Biotechnology Letters
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    • v.43 no.1
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    • pp.9-15
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    • 2015
  • The seaweed, Gracilaria verrucosa, was fermented to produce bioethanol. Optimal pretreatment conditions were determined to be 12% (w/v) seaweed slurry and 270 mM sulfuric acid at 121℃ for 60 min. After thermal acid hydrolysis, enzymatic saccharification was carried out with 16 U/ml of mixed enzymes using Viscozyme L and Celluclast 1.5 L to G. verrucosa hydrolysates. A total monosaccharide concentration of 50.4 g/l, representing 84.2% conversion of 60 g/l total carbohydrate from 120 g dw/l G. verrucosa slurry was obtained by thermal acid hydrolysis and enzymatic saccharification. G. verrucosa hydrolysate was used as the substrate for ethanol production by separate hydrolysis and fermentation (SHF). Ethanol production by Candida lusitaniae ATCC 42720 acclimated to high-galactose concentrations was 22.0 g/l with ethanol yield (YEtOH) of 0.43. Acclimated yeast to high concentrations of specific sugar could utilize mixed sugars, resulting in higher ethanol yields in the seaweed hydrolysates medium.

Enzyme Activities and Cellulose Degradation of Domestic Softwoods in Shaking Culture of Fomitopsis palustris (국내산 침엽수 목분의 진탕배양에서 나타난 Fomitopsis palustris의 효소 활성 및 셀룰로오스 분해)

  • Choi, Doo-Yeol;Lee, Young-Min;Kim, Young-kyoon;Yoon, Jeong-Jun;Kim, Yeong-Suk
    • Journal of the Korean Wood Science and Technology
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    • v.35 no.6
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    • pp.91-99
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    • 2007
  • Activities of the extracellular enzyme from Fomitopsis palustris, a brown-rot fungi, and by which crystallinity changes of cellulose in the various softwoods, such as Larix leptolepsis, Finns rigida, Finns koraiensis and Finns densiflora by liquid culture, were investigated. Activity of Cellobiohydrolase (CBH) from F. palustris was detected in the every test softwoods culture, showing activities of the Endoglucanase (EG), $\beta$-glucosidase (BGL) and $\beta$-1,4-xylosidase (BXL). It was shown high enzyme activities in the sapwood culture than heartwood of the same wood species, However, the enzyme activities in most of test wood cultures increased with longer incubation time, indicating a possibility of intermix sapwood and heartwood for degradation process by enzyme. Also it was shown that protein patterns of the extracellular enzyme from F. palustris in wood particle substrate of the several domestic softwoods were similar with each other wood species, which suggested the possibility of mixing all softwoods in saccharification by enzyme from F. palustris. Crystallinity reduction value of cellulose by F. palustris was 4.2~20.4% in 4 weeks cultivation, 12.9~28.9% in 8 weeks.

Study on the Relationships between Rice Straw Degradation and Changes of Fibrolytic Bacteria Population by in Vitro Rumen Fermentation (In Vitro 반추위 발효를 통한 볏짚 분해와 섬유소 박테리아 군집 변화의 관계 연구)

  • Sung, Ha Guyn
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.37 no.1
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    • pp.35-43
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    • 2017
  • This study was to research the relationships between rice straw degradation and changes of fibrolytic bacteria population during the in vitro rumen fermentation. Dry matter(DM) digestion of rice straw and population of fibrolytic bacteria were measured at the 0. 4, 8, 12 and 48 hours during the incubation. The populations of F. succinogenes. R. albus and R. flavefaciens were defined as log copy number of 16S rDNA by technical method of Quantitative real-time PCR. Total population of F. succinogenes, R. flavefaciens and R. albus was sum of bactera attached on rice straw and suspended in medium. It's population was increased with incubation, reached top level of 29.0 Log copy No at the 24 hour and then decreased. In the meantime, DM digestion of rice straw showed the higher increasement from the 8 hour to the 24 hour than from the 0 hour to the 8 hour, and then a slowdown in increasing trend of digestibility. Attachments of F. succinogenes, R. flavefaciens and R. albus were detected immediately after start of in vitro rumen incubation. At the same time, the colonized bacterial share were respectively 34.5%, 84.4% and 67.9% in total population. All of them was reached the highest colonized bacterial share above 94.7% at the 4 hour incubation. However population of attached bacteria was shown the highest level at the 12 hour or the 24 hour incubation. Kinetics of colonization were formed area of top speed from the 12 hour to the 24 hour and respectively reached 10.33, 9.28 및 8.30 Log copy No/h/g DM at the 24 hour by F. succinogenes, R. flavefaciens and R. albus. The kinetics of rice straw degradation was formed top level of 0.95% DM/h at the 24 hour. The present results gave clear evidence that degradation of rice straw was increased with the development of total fibrolytic bacteria in process of rumen fermentation. Also, their attachment was largely occurred immediately after insertion of rice straw, the colonized bacteria was actively proliferated, and then degradation of rice straw was maximized.

Effects of Added Vegetable Oils on In vitro Formation of Fatty Acid Soaps and Fermentation Characteristics and NDF Disappearance Rate (식물성유 첨가가 In vitro 발효성상, NDF 소실율 및 지방산염 형성에 미치는 영향)

  • Kim, D.I.;Choi, J.R.;Lee, Y.H.;Lee, J.K.;Chung, T.Y.
    • Journal of Animal Science and Technology
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    • v.46 no.3
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    • pp.355-372
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    • 2004
  • In vitro experiments were conducted to determine the formation of fatty acid soaps (FAS) and neutral detergent fiber (NDF) disappearance rate. The substrates were a basal alfalfa hay containing 1) no oil, 2) 10% soybean oil, 3) 10% com oil, on a weight basis. All the substrates were incubated in triplicate for 0, 3, 6, 12, 24 and 48h in each experiment. After the incubation in the first experiment serum bottles (6oml) were analyzed for nonesterified, esterifed and fatty acid soaps contents. The serum bottles (120mI) from the second experiment were analyzed for pH, $NH_3-N$ and VFA concentration, and dry matter and NDF disappearance rate. pH decreased and the concentration of NH3-N increased significantly with longer incubation time (P<0.0001). The disappearance rates of dry matter and NDF significantly varied with feed, incubation time and oils (P<0.05). The molar concentration of total VFA increased and proportion of acetate significantly decreased with incubation time (P<0.0001), but the proportion of propionate significantly increased with longer incubation time (P<0.0001). Addition of oils to diet lowered the ratio of acetate:propionate (P<0.05). The esterified fatty acids (EFA) decreased with increasing incubation time (P<0.0001), and nonesterified fatty acids (NEFA) increased due to lipolysis of EFA, NEFA then reacted with cations to increase formation of FAS. The formation of FAS increased significantly at 48h of incubation time (P<0.0001). Especially, formation of stearic acid soaps was 27.5 and 32.5 folds with soybean oil and com oil supplements, respectively, by 48h of incubation time (P<0.0001). Alfalfa hay had higher cation contents, particularly Ca, which react with NEFA and FAS can be formed with longer incubation time. Saturated fatty acids had a higher proportion of FAS than did unsaturated fatty acids, suggesting that the former may react more extensively with cations. FAS contents increased with increasing chain length of the fatty acids. Since added vegetable oils fonned FAS, it might decrease negative effects on in vitro fermentation characteristics and NDF disappearance rate.