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Studies on the Production of 5'-Nucleotides by Streptomyces spp. -Part 1. Isolation of 5'-phosphodiesterase Producing Microorganisms- (방사균(放射菌)에 의한 5'-Nucleotide류(類)의 생산(生産)에 관한 연구(硏究) -제 1 보(第 1 報) 5'-Phosphodiesterase생산균(生産菌)의 분리(分離)-)

  • Kim, Hong-Jip;Bae, Chong-Chan;Hwang, Kyu-Ln;Kong, Un-Young
    • Applied Biological Chemistry
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    • v.22 no.4
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    • pp.210-216
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    • 1979
  • RNA degrading bacteria were isolated from soil of Korea. One strain (no. JSC-114), having strong 5'-phosphodiesterase activity, was identified as belonging to the genus Streptomyces on the basis of taxonomic characteristics. The optimum conditions of 5'-phophosdiesterase production were found at $30^{\circ}C$ for 4 day in a medium containing 4.5% of soluble starch, 0.15% of peptone, 0.6% of yeast extract, 0.1% of $MgSO_4{\cdot}7H_2O$, 0.01% of $CaCl_2{\cdot}2H_2O$, 0.25% of $KNO_3$, and 0.5% of $KH_2PO_4$(pH 7.0). The maximum production rate of 5'-nucleotides from yeast RNA was 95% at $40-45^{\circ}C$ for 4hrs, and the products were identified as 5'-IMP, 5'-GMP, 5'-CMP and 5'-UMP(5.5 : 5.0 : 4.9 : 5.0).

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Elimination and Utilization of Pollutants - Part I Microbiological Clarification of Industrial Waste and Its Utilization as Feed Resources - (환경오염원(環境汚染源)의 제거(除去)와 그 이용성(利用性)에 관(關)한 연구(硏究) - 제(報I)1보(第). 미생물(微生物)에 의(依)한 산업폐수(産業廢水)의 정화(淨化) 및 사료자원개발(飼料資源開發)에 개(開)하여 -)

  • Lee, Ke-Ho;Lee, Kang-Heup;Park, Sung-O
    • Applied Biological Chemistry
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    • v.23 no.1
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    • pp.64-72
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    • 1980
  • Industrial wastes from pulp and food plants were treated with microorganisms to clarify organic waste-water and to produce cells as animal feed, and results were summarized as follows. (1) Waste-water from pulp, beer, bread yeast, and ethanol distillation plants contained $1.4{\sim}1.5%$ of total sugar, $0.25{\sim}0.35%$ nitrogen, and biological oxygen demand (BOD) was $400{\sim}25,000$, chemical oxygen demand (COD), $500{\sim}28,000$, and pH, $3.8{\sim}7.0$. The BOD and COD were highest in waste-water from ethanol distillation plants among others. (2) Bacterial and yeast counts were $4{\times}10^4-1{\times}10^9,\;2{\times}10^2-7{\times}10^4/ml$ in waste-water. (3) Bacteria grew better in pulp waste and yeasts in beer, bread yeast, and ethanol distillation waste. (4) Saccharomyces cerevisiae SAFM 1008 and Candida curvata SAFM 70 were the most suitable microorganisms for clarification of ethanol distillation waste. (5) When liquid and solid waste from ethanol distillation were treated with microbial cellulase, xylanase, and pectinase, solid waste was reduced by 36%, soluble waste was increased, and recuding sugar content was increased by 1.3 times which provided better medium than untreated waste for cultivation of yeasts. (6) Optimum growth conditions of the two species of yeast in ethanol distillation waste were pH 5.0, $30^{\circ}C$, and addition of 0.2% of urea, 0.1% of $KH_2PO_4$ and 0.02% of $MgSO_4$. (7) Minimum number of yeast for proper propagation was $1.8{\times}10^5/ml$. (8) C. curvata70 was better than cerevisae for the production of yeast cells from ethanol distillation waste treated with microbial enzymes. (9) S. cerevisiae produced 16 g of dried cell per 1,000ml of ethanol distillation waste and reduced BOD by 46%. C. curvata produced 17.6g of dried cell and reduced BOD by 52% at the same condition. (10) Yeast cells produced from the ethanol distillation waste contained 46-52% protein indicating suitability as a protein source for animal feed.

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A New Synthetic Medium for Lactic Lactococci: Application to Marine lactic Acid Bacteria

  • KIM Joong K.;BAJPAI Rakesh K.
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.28 no.6
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    • pp.812-813
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    • 1995
  • Lactococcal cells are nutritionally fastidious and thus, generally cultured either in milk or M17 medium (Terzaghi and Sandine, 1975). In this study, Lactococcus cremoris wild-type (KH) and its less­proteolytic mutant (KHA1) cells were grown on the M17 medium or with modified M17 medium by replicated parallel experiments. The modified M17 medium had the same composition as M17 medium, except that lactose was replaced by glucose. Analyses of culture-broth samples, in which the M17 and the modified M17 media were used, were conducted by high-performance liquid chromatography (HPLC). But, working with these media created noisy problems in analyses of samples. Therefore, a new semi-synthetic medium was developed on the basis of nutritional requirements (Morishita et al., 1981). The composition of the semi-synthetic medium determined on the basis of the nutritional requirements and the composition of milk, is presented in Table 1. The composition of M17 medium is also presented and compared in the table. L. cremoris KH and KHA1 cells were grown again on the new synthetic medium containing glucose or lactose. The broth samples were then drawn and analyzed by HPLC. Clearer separations of fermented products were achieved from the new medium than those with the M17 and the modified M17 media. In comparison with the M17 or the modified M17 media, growth on the new medium was good (Kim et al, 1993). Additional fermentations were also carried out at a controlled pH of 7.0, where enhanced growth of lactococcal cells was obtained. In the fermentations, samples were also analyzed for the concentrations of sugar and lactic acid. The results showed that the new synthetic medium was as good as or better than the M 17 and the modified M 17 media. This is because casein hydrolysate in the synthetic medium provided a ready supply of amino acids and peptides for L. cremoris KH and KHA1 cells. Lactic acid bacteria (LAB) including Lactococcal cells have been known to be an effective means of preserving foods, at the same time as giving particular tastes in fields of dairy products. LAB also have always occupied an important place in the technology of sea products, and marine LAB have known to be present in traditional fermented products (Ohhira et al, 1988). To apply the new synthetic medium to marine LAB, two different LAB were isolated from pickled anchovy and pollacks caviar and were grown on the new media in which various concentrations of NaCl $(3, 5, 7 and 10\%)$ added. They were also grown on the medium solution in natural seawater $(35\%o\;salinity)$ and on the solution of natural seawater itself, too. As seen in Fig. 1, Marine LAB were grown best on the synthetic medium solution in natural seawater and the higher concentrations of NaCl were added to the medium, the longer lag-phase of growth profile appeared. Marine LAB in natural seawater were not grown well. From these results, the synthetic medium seems good to cultivate cells which are essential to get salted fish aged. In this study, it showed that the new synthetic medium provided adequate nutrition for L. cremoris KH and KHA1 cells, which have been used as cheese starters (Stadhouders et al, 1988). Using this new medium, the acid production capability of starter cultures could be also measured quantitatively. Thus, this new medium was inferior to the M17 or the modified M17 medium in culturing the cheese starters and in measuring fermentation characteristics of the starter cells. Moreover, this new medium found to be good for selected and well-identified marine LAB which are used in rapid fermentations of low-salted fish.

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Physiological Responses to Mineral-Excessive Conditions: Mineral Uptake and Carbohydrate Partitioning in Tomato Plants

  • Sung, Jwakyung;Lee, Suyeon;Lee, Yejin;Kang, Seongsoo;Ha, Sangkeun;Sonn, Yeonkyu
    • Korean Journal of Soil Science and Fertilizer
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    • v.47 no.6
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    • pp.563-570
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    • 2014
  • The shortage or surplus of minerals directly affects overall physiological metabolism of plants; especially, it strongly influences carbohydrate metabolism as a primary response. We have studied mineral uptake, synthesis and partitioning of soluble carbohydrates, and the relationship between them in N, P or K-excessive tomato plants, and examined the interaction between soluble carbohydrates and mineral elements. Four-weeks-old tomato plants were grown in a hydroponic growth container adjusted with excessive N ($20.0mmol\;L^{-1}$ $Ca(NO_3)2{\cdot}4H_2O$ and $20.0mmol\;L^{-1}$ $KNO_3$), P ($2.0mmol\;L^{-1}$ $KH_2PO_4$), and K ($20.0mmol\;L^{-1}$ $KNO_3$), respectively, for 30 days. Shoot growth rates were significantly influenced by excessive N or K, but not by excessive P. The concentrations of water soluble N (nitrate and ammonium), P and K were clearly different with each tissue of tomato plants as well as the mineral conditions. The NPK accumulation in all treatments was as follows; fully expanded leaves (48%) > stem (19%) = roots (16%) = petioles (15%) > emerging leaves (1). K-excessive condition extremely contributed to a remarkable increase in the ratio, which ranged from 2.79 to 10.34, and particularly potassium was dominantly accumulated in petioles, stem and roots. Fresh weight-based soluble sugar concentration was the greatest in NPK-sufficient condition ($154.8mg\;g^{-1}$) and followed by K-excessive (141.6), N-excessive (129.2) and P-excessive (127.7); whereas starch was the highest in K-excessive ($167.0mg\;g^{-1}$) and followed by P-excessive (146.1), NPK-sufficient (138.2) and N-excessive (109.7). Soluble sugar showed positive correlation with dry weight-based total N content (p<0.01) whereas was negatively correlated with soluble P (p<0.01) and dry weight-based total P (p<0.01). On the other hand, starch production was negatively influenced by total N (p<0.001), but, it showed positive relation with total K concentration (p<0.05). This study shows that uptake pattern of NPK and production and partitioning of soluble carbohydrate were substantially different from each mineral, and the relationship between water soluble- and dry weight-based-mineral was positive.

High Cell Density Culture of Micro-algal Dunaliella bardawil (미세조류 Dunaliella bardawil의 고농도 세포배양)

  • 정욱진;왕만식;최승인;정병철;김주곤
    • KSBB Journal
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    • v.14 no.2
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    • pp.160-166
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    • 1999
  • High cell density cultivation of microalga Dunaliella bardawil using nitrogen fed-batch cultures was studied in batch flask. Optimum environmental conditions include concentrated nutrients except NaCl and carbon sources, carbon sources, pH, light, agitation, nitrate and phosphate ions. Cell growth, consumption rates of nitrate and phosphate ions were monitored. Optimal conditions for higher cell density were found to be(in the range tested): 5 times concentrated media(1 times-10 times concentrated media) pH 8.0 (7.0-9.0) white light(blue and red light) 15mM of nitrate (0.94-15mM) 250mM $NaHCO_3$ and $CO_2$ gas. However, the addition of phosphate ions did not enhance the algal maximum cell density and specific growth rate. Nitrate was found to be effective for the cell growth. The maximum cell density of fed-batch culture using nitrate ions in $8.955{\times}106$cells/ml after 189hr incubation.

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Production and Characterization of Keratinase from Paracoccus sp. WJ-98

  • Lee, Yoon-Jeong;Kim, Jae-Ho;Kim, Ha-Kun;Lee, Jong-Soo
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.1
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    • pp.17-22
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    • 2004
  • A bacterial strain WJ-98 found to produce active extracellular keratinase was isolated from the soil of a poultry factory. It was identified as Paracoccus sp. based on its 16S rRNA sequence analysis, morphological and physiological characteristics. The optimal culture conditions for the production of keratinase by Paracoccus sp. WJ-98 were investigated. The optimal medium composition for keratinase production was determined to be 1.0% keratin, 0.05% urea and NaCl, 0.03% K$_2$HPO$_4$, 0.04% KH$_2$PO$_4$, and 0.01% MgCl$_2$$.$6H$_2$O. Optimal initial pH and temperature for the production of keratinase were 7.5 and 37$^{\circ}C$, respectively. The maximum keratinase production of 90 U/mL was reached after 84 h of cultivation under the optimal culturing conditions. The keratinase from Paracoccus sp. WJ-98 was partially purified from a culture broth by using ammonium sulfate precipitation, ion-exchange chromatography on DEAE-cellulose, followed by gel filtration chromatography on Sephadex G-75. Optimum pH and temperature for the enzyme reaction were pH 6.8 and 50$^{\circ}C$, respectively and the enzymes were stable in the pH range from 6.0 to 8.0 and below 50$^{\circ}C$. The enzyme activity was significantly inhibited by EDTA, Zn$\^$2+/ and Hg$\^$2+/. Inquiry into the characteristics of keratinase production from these bacteria may yield useful agricultural feed processing applications.

Molecular Cloning of nod Genes from Bradyrhizobium sp. SNU001 (Bradyrhizobium sp. SNU001 nod 유전자 클로닝)

  • 고세리;심웅섭;안정선
    • Korean Journal of Microbiology
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    • v.30 no.4
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    • pp.246-251
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    • 1992
  • Molccular cloning of nod genes from Bradvrhizobium sp. SNU001, a nitrogen-fixing symbiont isolated from thc root nodules of soybean (Clycine trim) . was carried out. nod genes were found to be located on thc genome of the symbiont by gcnomic hybridization with 4.5 kb EcoRI/HndIII fragment (nod DABC) of Rhizohium meliloti as probe. Genomic library of this symbiont was constructed using h phage EMBL3-BanlHI vector. from which five nod positive clones were sclectcd by primary and secondary screening methods. The partial restriction map of inserted genomic DNA of h CNS-l(c1one 2) was constructed. and 3.9 kh Bun7HI fragment. which showed strong hybridization signal to the probe, was subcloned into pBS KS(+) plasmid vector. Partial restriction inap ot' a selected subclone (pBjCNS-I) was constructed and nod DABC was found to be located on the 1.8 kb KpnI/Sacl fragment of this subclone.

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Effects of Supplemental Phosphorus and Iron in the Fish Meal Diets on Growth in Juvenile Rockfish (Sebastes schlegeli) (단백질원이 어분인 조피볼락 치어 사료에 인(P)과 철(Fe)의 보충 효과)

  • 박승렬;이상민;정관식;장영진
    • Journal of Aquaculture
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    • v.10 no.3
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    • pp.255-260
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    • 1997
  • In order to investigate the effects of phosphorus and iron levels in the diets containing fish meal as only protein source on the growth, feed conversion, body composition and blood characteristics of juvenile Korean rockfish, either graded monobasic potassium phosphate (MPP) of 1% (MPP-1), 2% (MPP-2), 3% (MPP-3) and 4% (MPP-4), or Fe-citrate (FC) of 0.022% (FC-1), 0.044% (FC-2), 0.066% (FC-3) and 0.088% (FC-4) was added to fish meal-based diet (MMP-0 or FC-0), respectively. Duplicate groups of 40 fish weighing 7g were fed the diets for 21 weeks. Weight gain, feed efficiency and nutrient retention of fish fed diets with different MPP levels were not significantly different (P>0.05). Lipid content of whole body decreased with an increase in dietary MMP (P<0.05). No significant differences were found in the Ht, Hb, total protein, or total glucose in serum among diets containing different MMP levels (P>0.05). Weight gain of fish fed the FC-0, FC-1 adn FC-3 diets was significantly higher than that of FC-4 diet (P>0.05), whereas feed efficiency, nutrient retention, hematological values (Ht, Hb, total protein, and total glucose), and whole body composition were not affected by dietary FC levels (P>0.05). The present results revealed that phosphate or iron supplementation to the diet containing 65% fish meal for juvenile Koran rockfish was not necessary.

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Fruit-body Formation of Pleurotus florida on the Synthetic Medium (합성배지(合成培地)를 이용(利用)한 고온성(高溫性) 느타리버섯의 자실체(子實體) 형성(形成)에 관한 연구(硏究))

  • Hong, Jai-Sik;Kang, Kui-Hwan
    • The Korean Journal of Mycology
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    • v.11 no.3
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    • pp.121-128
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    • 1983
  • Nutritional and physio-chemical conditions for mycelial growth and fruit-body formation of Pleurotus florida were determined in synthetic media. Mannitol and sucrose were good sugar substances for the mycelial growth and fruit-body formation whereas less mycelial growth and no fruit-body formation was obtained with arabinose, lactose and inulin. The optimum concentration of mannitol was about 2%. Peptone as a nitrogen sources resulted in a rapid mycelial growth and fruit-body formation with higher yield, but nitrite-nitrogens inhibited the mycelial growth. The higher yield was obtained with 0.2% peptone. Among the vitamins used, the greatest mycelial growth and fruit-body formation brought about by thiamine and folic acid. $KH_2PO_4$ and $MgSO_4$ at 0.2% and 0.02%, respectively, were effective for the mycelial growth and fruit-body formation, but other inorganic salts used were ineffective. The optimum temperature for mycelial growth and fruit-body formation were $25^{\circ}C$ and $20^{\circ}C$, respectively, and light intensity of $100{\sim}500\;lux$ and pH 6.0 appeared to be effective.

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Optimization of Medium Composition for Lipopeptide Production from Bacillus subtilis N7 using Response Surface Methodology

  • Luo, Yi;Zhang, Guoyi;Zhu, Zhen;Wang, Xiaohui;Ran, Wei;Shen, Qirong
    • Microbiology and Biotechnology Letters
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    • v.41 no.1
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    • pp.52-59
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    • 2013
  • The nutritional requirements for the maximum production of lipopeptides by Bacillus subtilis N7 (B. subtilis N7) were investigated and optimized using response surface methodology (RSM) under shake flask fermentation. A one-factor-at-a-time experimental setup was used to screen carbon and nitrogen sources. A Plackett-Burman design (PBD) was employed to screen the most critical variables for lipopeptides production amongst ten nutritional elements. The central composite experimental design (CCD) was finally adopted to elucidate the composition of the fermentation medium. Statistical analyses (analysis of variance, ANOVA) of the results showed that KCl, $MnSO_4$ and $FeSO_4{\cdot}6H_2O$ were important components and that their interactions were strong. Lipopeptide production was predicted to reach 709.87 mg/L after a 60 h incubation using an optimum fermentation medium composed of glucose 7.5 g/L, peanut oil 1.25 g/L, $MgSO_4$ 0.37 g/L, $KH_2PO_4$ 0.75 g/L, monosodium glutamate 6.75 g/L, yeast extract and $NH_4Cl$ (5:3 w/w) 10 g/L, KCl 0.16 g/L, $FeSO_4{\cdot}6H_2O$ 0.24 mg/L, $MnSO_4$ 0.76 mg/L, and an initial pH of 7.0. Lipopeptide production ($706.57{\pm}3.70$ mg/L) in the optimized medium confirmed the validity of the predicted model.