• Title/Summary/Keyword: Optimum Strain Rate

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Isolation of a Phenol-degrading Bacterial Strain and Biological Treatment of Wastewater Containing Phenols (Phenol 분해균주의 분리 및 페놀함유 폐수의 생물학적 처리)

  • Lee, Hyun Don;Lee, Myoung Eun;Kim, Hyung Gab;Suh, Hyun-Hyo
    • Journal of Life Science
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    • v.23 no.10
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    • pp.1273-1279
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    • 2013
  • Aromatic hydrocarbons, such as phenol, have been detected frequently in wastewater, soil, and groundwater because of the extensive use of oil products. Bacterial strains (56 isolates) that degraded phenol were isolated from soil and industrial wastewater contaminated with hydrocarbons. GN13, which showed the best cell growth and phenol degradation, was selected for further analysis. The GN13 isolate was identified as Neisseria sp. based on the results of morphological, physiological, and biochemical taxonomic analyses and designated as Neisseria sp. GN13. The optimum temperature and pH for phenol removal of Neisseria sp. GN13 was $32^{\circ}C$ and 7.0, respectively. The highest cell growth occurred after cultivation for 30 hours in a jar fermentor using optimized medium containing 1,000 mg/l of phenol as the sole carbon source. Phenol was not detected after 27 hours of cultivation. Based on the analysis of catechol dioxygenase, it seemed that catechol was degraded through the meta- and ortho-cleavage pathway. Analysis of the biodegradation of phenol by Neisseria sp. GN13 in artificial wastewater containing phenol showed that the removal rate of phenol was 97% during incubation of 30 hours. The removal rate of total organic carbon (TOC) by Neisseria sp. GN13 and activated sludge was 83% and 78%, respectively. The COD removal rate by Neisseria sp. GN13 from petrochemical wastewater was about 1.3 times higher than that of a control containing only activated sludge.

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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Effects of Yeast Strains and Fermentation Temperatures in Production of Hydrogen Sulfide During Beer Fermentation (맥주의 발효과정에서 효모와 발효온도가 황화수소의 발생에 미치는 영향)

  • Kim, Young-Ran;Moon, Seung-Tae;Park, Seung-Kook
    • Korean Journal of Food Science and Technology
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    • v.40 no.2
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    • pp.238-242
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    • 2008
  • In this study, hydrogen sulfide ($H_2S$) production was examined during beer fermentation using two ale and two lager yeast strains. In the lager yeast fermentation, a large amount of $H_2S$ was produced in the early fermentation stages when the yeast were actively fermenting wort, indicating a positive relationship between the level of H2S production and the yeast growth rate during fermentation. The ale yeasts produced much lower levels of H2S than the lager yeasts. In the lager fermentation, a higher fermentation temperature shortened the fermentation period, but much higher levels of $H_2S$ were produced at higher temperatures. American pilsner lager yeast fermenting at $15^{\circ}C$ produced a relatively high level of $H_2S$ at the end of fermentation, which would require a longer aging time to remove this malodorous volatile sulfur compound. Not including the English ale strain, which produced a higher level of H2S at lower temperatures, the ale yeast produced lower levels of $H_2S$ at lower temperatures, suggesting that each strain has an optimum fermentation temperature for H2S production.

The Study on Recombinant Protein Production using S. cerevisiae Mutant Y334 Suitable for GAL Promoter (GAL promoter에 적합한 효모변이주 Y334의 회분식 배양에서의 재조합 단백질 발현특성)

  • Gang, Hwan-Gu;Lee, Mun-Won;Jeon, Hui-Jin
    • KSBB Journal
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    • v.14 no.4
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    • pp.476-481
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    • 1999
  • S. cerevisiae mutant(reg1-501, gal1), which cannot use galactose and has alleviated glucose repression level, is used as host for optimizing induction of GAL promoter. The optimum concentration of galactose as inducer for recombinant protein production and the galactose consumption rate have been tested with S. cerevisiae mutant and compared with conventional S. cerevisiae. The extent of glucose repression were investigated for both strain and the degradation pattern of produced foreign protein have been compared in both cases. The effect of pH on foreign protein degradation pattern were studied for both strains. The secetion efficiency of both strains were carried out. Through these experiments, optimum condition of recombinant protein production by GAL promoter using S. cerevisiae mutant (reg1-501, gal1) were found.

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Studies on the Bacteriophages of Brevibacterium lactofermentum (L-글루타민산 생산균 Brevibacterium lactofermentum의 Bacteriophag에 관한 연구)

  • 이태우
    • Korean Journal of Microbiology
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    • v.17 no.3
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    • pp.97-130
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    • 1979
  • Many industrial processes those employ bacteria are subjected to phage infestations. In L-glutamic acid fermentions using acetic acid, the phage infestations of the organisms have been recently recognized. In efforts to elucidate the sources of phage contamination involved in the abnormal fermentation, a series of study was conducted to isolate the phages both from the contents of abnormally fermented tanks and the soil or sewage samples from the surroundings of a fermentation factory, to define major charateristics of the phage isolates, and finally to determine the correlation between the phage isolates and temperate phages originating from the miscellaneous bacterial species isolated from the soil or sewage samples. The results are summarized as follows; 1) All phages were isolated from the irregular fermentation tanks and soil or sewage samples, and they were designated as phage PR-1, PR-2, PR-3, PR-4, PR-5, PR-6, and PR-7, in the order of isolation. These PR-series phages were proved to be highly specific for the variant strains of Br. lactofermentum only, namely, phage PR-1 and PR-2 for Br. lactofermentum No. 468-5 and phage PR-3~PR-7 for Br. lactofemrentum No. 2256. By cross-neutralization test, the 7 phagescould be subdivided into 3 groups, i. e., phage PR-I and PR-2 the first, phage PR-3, PR-4, PR-5, PR-6 the second, and the phage PR-7 the third. 2) The 7 phages were virulent under the experimental conditions. They produced plaques with clear and relatively sharp margins without distinct halo. The mean sizes of plaques were 1.5mm in diameter for phage PR-1 and PR-2, and 1. Omm for phages PR-3~PR-7. Double layer technique modified by Hongo and described by Adams, was applied to assay of the PR-series phages. The factors influencing the plaques were as follows;young age cells of host bacteria cultured for 3-6 hours represented the largest number and size, optimum was pH 7.0, incubation temperature was $30^{\circ}C$, and agar concentration and amount of overlayer medium were 0.6% and 0.2ml, respectively. 3) PR-series phages were stable in 0.05M tris buffer and 0.1M ammonium acetate buffer solution. The addition of $5{\times}10^{-3}M$ magnesium ion effectively increased the stability. Thermostability experiments indicated that PR-series phages were stable at the teinperture between $50^{\circ}{\sim}55^{\circ}C$ in nutrient medium, $45^{\circ}{\sim}50^{\circ}C$ in buffer solution. However, the phages mere completely inactivated at 603C and 65$^{\circ}$C within 10 minutes. The phages were stable at the range of pH6~9 in nutrient medium and of pH 8-9 in buffer solution, respectively. Exposure of the phages to UV for 25, 60 and 100 seconds resulted in the complete loss of infectivily, respectively. 4) Electron microscopy showed that PR-series phage particles exhibited rather similar morphology, differing in the size All of PR-series phages had a multilateral head and had a simple long tiil about three to five times long as compared with head. By the size, phage PR-1 and PR-2, PR-3, PR-4, PR-5, and PR-6 and PR-7 were classified into same groups, respectively. The head and tail size of phage PR-1, PR-5, PR-5(T) and PR-7 were 85nm, 74nm and 235nm and 350mm, and 72nm and 210nm, respectively. 5) Nucleic acids of PR-series phages were double stranded DNA. The G+C contents of phage PR-1, PR-5 and PR-7 were 56.1, 52.9 and 53.7, respectively. The values of G+C contents derived from the $T_m$ were in agreement with the chemically determined values. 6) PR-series phages effectively adsorbed on their host bacteria at the rate of more than 90% during 5 min. K value for phage PR-1, PR-5 and PR-7 were calculated to be $6{\times}10^9 ml$ per minute, respectiveky. The pH of the medium did effect adsorption rate, but both temperature and age of host cells did not. Generally, optimum adsorption condition of phages seemed to be almost same as optimum growth conditions of host bacteria. 7) In one-step growth experiments, the latent periods at $30^{\circ}C$ for PR-1, and PR-7 were about 70, 50 and 55 min, respectively. The corresponding average burst size was 200, 70 and 90, respectively. Lpsis period according to the multiplicity of infection and a phage series. In case of m. o. i. 100, strain No. 2256 (PR-5) and No. 468-5(PR-1) failed to grow and turbidity decreased after 50 and 70min, respectively. 8) In the lysate of a plaque purified phage PR-5 infected bacteria, there observed 2 types ofphage particles, i. e., phage PR-5 and PR-5 (T) of similar morphology but differing at the length of phage tail, and phage tail like particles. The phage taillike particles could be divided into 4 types by the length. Induction experiments of Br. lactofermentum with UV irradiation, mitomycin C or bacitracin treatment produced neither phage PR-5 (T) or phage tail-like particles. 9) No lysis occured when the growth of 7 strains of miscellaneous bacteria, isolated from soil and sewage samples, were inoculated with either phage PR-5 (T) or phage tail-like particles the inoculation of phage PR-5 pellet resulted in the growth inhibition of the orgainsms in the spot test. The lysates obtained from 3 miscellaneous soil derived bacteria following mitomycin C treatment the growth of Br. lactofermentum, but did not lyze the bacterium.

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Importance of Strain Improvement and Control of Fungal cells Morphology for Enhanced Production of Protein-bound Polysaccharides(β-D-glucan) in Suspended Cultures of Phellinus linteus Mycelia (Phellinus linteus의 균사체 액상배양에서 단백다당체(β-D-glucan)의 생산성 향상을 위한 균주 개량과 배양형태 조절의 중요성)

  • Shin, Woo-Shik;Kwon, Yong Jung;Jeong, Yong-Seob;Chun, Gie-Taek
    • Korean Chemical Engineering Research
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    • v.47 no.2
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    • pp.220-229
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    • 2009
  • Strain improvement and morphology investigation in bioreactor cultures were undertaken in suspended cultures of Phellinus linteus mycelia for mass production of protein-bound polysaccharides(soluble ${\beta}$-D-glucan), a powerful immuno-stimulating agent. Phellineus sp. screened for this research was identified as Phellinus linteues through ITS rDNA sequencing method and blast search, demonstrating 99.7% similarity to other Phellinus linteus strains. Intensive strain improvement program was carried out by obtaining large amounts of protoplasts for the isolation of single cell colonies. Rapid and large screening of high-yielding producers was possible because large numbers of protoplasts ($1{\times}10^5{\sim}10^6\;protoplasts/ml$) formed using the banding filtration method with the cell wall-disrupting enzymes could be regenerated in relatively high regeneration frequency($10^{-2}{\sim}10^{-3}$) in the newly developed regeneration medium. It was demonstrated that the strains showing high performances in the protoplast regeneration and solid growth medium were able to produce 5.8~6.4%(w/w) of ${\beta}$-D-glucan and 13~15 g/L of biomass in stable manners in suspended shake-flask cultures of P. linteus mycelia. In addition, cell mass increase was observed to be the most important in order to enhance ${\beta}$-D-glucan productivity during the course of strain improvement program, since the amount of ${\beta}$-D-glucan extracted from the cell wall of P. linteus mycelia was almost constant on the unit biomass basis. Therefore we fully investigated the fungal cell morphology, generally known as one of the key factors affecting cell growth extent in the bioreactor cultures of mycelial fungal cells. It was found that, in order to obtain as high cell mass as possible in the final production bioreactor cultures, the producing cells should be proliferated in condensed filamentous forms in the growth cultures, and optimum amounts of these filamentous cells should be transferred as active inoculums to the production bioreactor. In this case, ideal morphologies consisting of compacted pellets less than 0.5mm in diameter were successfully induced in the production cultures, resulting in shorter period of lag phase, 1.5 fold higher specific cell growth rate and 3.3 fold increase in the final biomass production as compared to the parallel bioreactor cultures of different morphological forms. It was concluded that not only the high-yielding but also the good morphological characteristics led to the significantly higher biomass production and ${\beta}$-D-glucan productivity in the final production cultures.

Development of Saccharomyces cerevisiae Strains with High RNA Content (리보핵산을 다량으로 함유하는 Saccharomyces cerevisiae 균주의 개발)

  • Kim, Jae-Sik;Kim, Jin-Wook;Shim, Won;Min, Byoung-Cheol;Kim, Jung-Wan;Park, Kwan-Hwa;Pek, Un-Hua
    • Korean Journal of Food Science and Technology
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    • v.31 no.2
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    • pp.465-474
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    • 1999
  • RNase activity of Saccharomyces cerevisiae ATCC 7754 was investigated to obtain strains with high ribonucleic acid (RNA) content. The yeast strain contained two RNase activities; an acidic RNase with a optima of pH $3{\sim}4$ and an alkaline RNase with a optima pH 9. The acidic RNase activity was inhibited by $0.08\;M\;HgCl_{2}$ most drastically. The alkaline RNase activity was inhibited by 2.0 M NaCl or KCl, while enhanced by addition of $0.05\;M\;CaCl_{2},\;0.02\;M\;ZnSO_{4},\;or\;0.008\;M\;HgCl_{2}$. Various mutants of Saccharomyces cerevisiae ATCC 7754 were isolated by ethylmethane sulfonate (EMS) treatment or $\gamma$-ray/ultra violet irradiation. Among the mutants that were sensitive to high concentration of KCl which inhibits alkaline RNase, B24 was selected for high RNA content per culture volume. Growth characteristics of the mutant were comparable to those of the mother strain with optimum growth at pH $4.5{\sim}5.5$. The mutant accumulated higher content of RNA than the mother strain when glucose was used as the carbon source. However, both growth rate and total RNA content of the mutant were higher in molasses medium than in glucose medium. RNA content of the mutant increased rapidly during the early stage of growth, and then decreased gradually until the culture reached stationary phase by a fed-batch culture in a 5 L jar fermenter. Maximal cell harvest and the final RNA content using the mutant B24 were 69.6 g/L culture broth and 19.8 g/100 g of the dry cell while those using the mother strain were 68 g/L culture broth and 16.1 g/100 g of dry cell, respectively.

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Production of L-α-Glycerophosphate Oxidase by streptococcus faecium M74 · LC (Streptococcus faecium M74 · LC에 의한 L-α-Glycerophosphate Oxidase의 생산)

  • Lee, In-Ae;Lee, Eun-Sook;Lee, June-Woo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.32 no.5
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    • pp.684-688
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    • 2003
  • The objective of this study was to search for the best strain as a source of L- $\alpha$-glycerophosphate oxidase (GPO) production and to establish the process technology for the purification of GPO on an industrial scale. The GPO was produced by culturing Streptococcus faecium, and purified by ammonium sulfate, DEAE-cellulose and hydroxyapatite chromatography. The relative activity was 60 units/L for 5. faecim ATCC 12755, 65 units/L for 5. faecium ATCC 19634, and 67 units/L for 5. faecium $M_{74}$.LC, respectively. The optimum condition for fermentation was $37^{\circ}C$ for temperature, 300 rpm for stir rate, 0.5 L/min for aeration rate and 17 hours. The main culture medium prepared by the modified AC medium. AC medium consists of 0.1% glucose, 0.2% glycerol, 1.0% tryptone and 1.0% yeast extract, 0.5% $K_2HP0_4$, pH 7.0. The GPO was purified by ammonium sulfate fractionation and ion exchange column chromatography, The yield and purity were 17.2% and 5.3 fold, respectively.

Temperature and dose-size effects on infectivity and reproduction of entomopathogenic nematode, Steinernema longicaudum Gongju Strain (온도와 농도가 곤충병원성 선충 Steinernema longicaudum 공주계통의 병원성과 증식에 미치는 영향)

  • Choo, Ho-Yul;Lee, Dong-Woon;Ha, Pan-Jung;Kim, Hyeong-Hwan;Chung, Hye-Jin;Lee, Sang-Myeong
    • The Korean Journal of Pesticide Science
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    • v.3 no.2
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    • pp.60-68
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    • 1999
  • Effects of temperature and dose-size on infectivity and reproduction of Korean entomopathogenic nematode, Steinernema longicaudum Gongju strain were examined. The greater wax mea Galleria mellonella larvae were exposed to 5, 10, 20, 40, 80, and 160 infective juveniles/larva in $60{\times}15$ mm petri dishes and kept in $13^{\circ}C$, $18^{\circ}C$, $24^{\circ}C$, and $30^{\circ}C$ incubators. Each petri dish contained one larva weighed from 180 to 200 mg. Infectivity was observed everyday for 14 days and reproduction for 30 days. The infectivity of S. longicaudum was more influenced by temperature than by dose-size. Mortalities by S. longicaudum were lower at $13^{\circ}C$ at all concentrations but higher at $24^{\circ}C$ and $30^{\circ}C$ even at lower concentrations, 5 or 10 infective juveniles/larva. Lethal time was also shorter with increasing temperature and dosages. All host larvae died at $24^{\circ}C$ and $30^{\circ}C$ in 2 days at the rate of 160 infective juveniles per host while 83.3% of tested larvae died at $24^{\circ}C$ in 10 days and 90% at $30^{\circ}C$ in 6 days at the rate of 5 infective juveniles. Reproduction was also better with increasing temperature and dosages. The highest number of progenies was obtained at $30^{\circ}C$ in 6 days at the rate of 80 infective juveniles. However, progenies were not produced from cadavers at $13^{\circ}C$. Reproductive period was the shortest at $30^{\circ}C$ of all temperatures by 6 to 9 days. The results indicated that optimum temperatures for infectivity was $24^{\circ}C$ and $30^{\circ}C$ for reproduction.

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Fermentation Process Characteristics of Phaffia rhodozyma Mutant B76 for Astaxanthin Biosynthesis (Astaxanthin 생합성을 위한 Phaffia rhodoxyma 변이주 B76의 발효공정 특성)

  • 임달택;이은규
    • KSBB Journal
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    • v.15 no.2
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    • pp.125-133
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    • 2000
  • Specific carotenoids and astaxanthin biosynthesis power of Phaffia rhodozyma mutant 876, which was obtained after NTG a and UV treatments, was higher than those of the wild type by 40% and 50%, respectively. The mutant strain did not show t the catabolite repression even at 22% (w/v) glucose concentration. The optimum C{N ratio was 2.0, and the optimum t temperature and initial pH were $22^{\circ}C$ and 6.0, respectively. 80th cell growth and astaxanthin formation decreased drastically a as the fermentation temperature was increased over $22^{\circ}C$, whereas they were comparable in the pH range between 5.0 and 7 7.0. Inoculum size did not affect the final cell density nor the carotenoids biosynthesis, and 3%(v/v) was selected as optimal. H Higher dissolved oxygen concentration facilitated astaxanthin biosynthesis, and aeration rate of 1.0 v/0/m and agitation speed of 400 rpm were selected as optimum. The final cell dens때 of 43.3 g/L and the volumetric astaxanthin and carotenoids concentrations of 110.6 mg/L and 149.4 mg/L, respectively, were obtained. The specific carotenoids concentration was 3.45 m mg{g-yeast(dry). Yx/s and Yp/s values of 0.37 and 1.08 were obtained. The result of this study will provide basic information u useful for mass production of astaxanthin from P. rhodozyma fermentation.

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