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Strain Improvement and Bioprocess Optimization for Enhanced Production of Haluronic Acid(HA) in Bioreactor Cultures of Streptococcus zooepidemicus (히알루론산 생산성 향상을 위한 Streptococcus zooepidemicus 균주 개량 및 발효조 배양공정 최적화)

  • Kim, Soo Yeon;Chun, Gie-Taek
    • Microbiology and Biotechnology Letters
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    • v.48 no.3
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    • pp.344-357
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    • 2020
  • Strain improvement and bioprocess development were undertaken to enhance hyaluronic acid(HA) production by Streptococcus zooepidemicus cells. Using a high-yielding mutant strain, statistical medium optimization was carried out in shake flask cultures, resulting in 52% increase in HA production (5.38 g/l) at the optimal medium composition relative to the parallel control cultures. For sufficient supply of dissolved oxygen (DO), which turned out to be crucial for enhanced production of HA, agitation system and speed were intensively investigated in 5 L bioreactor cultures. Increase in oxygen mass transfer coefficient (kLa) through increment of agitation speed (rpm) and 35% expansion of diameter of the newly-designed impellers showed significantly positive effects on HA production. By installing an expanded Rushton-turbine impeller for efficient break-down of sparged air, and an extended marine impeller above the Rushton-turbine impeller for efficient mixing of the air-born viscous fermentation broth, maximum amount of HA (9.79 g/l) was obtained at 450 rpm, 1.8 times higher level than that of the corresponding flask culture. Subsequently, the possibility of bioprocess scale-up to a 50 L bioreactor was investigated. Despite almost identical maximum HA production (9.11 vs 9.25 g/l), the average HA volumetric productivity (rp) of the 50 L culture turned out only 74% compared to the corresponding 5 L culture during the exponential phase, possibly caused by shear damages imposed on the producing cells at the high stirring in the 50 L culture. The scale-up process could be successfully achieved if a scale-up criterion of constant oxygen mass transfer coefficient (kLa) is applied to the 50 L pilot-scale bioreactor system.

Optimization of Production Medium by Response Surface Method and Development of Fermentation Condition for Monascus pilosus Culture (Monascus pilosus 배양을 위한 반응표면분석법에 의한 생산배지 최적화 및 발효조건 확립)

  • Yoon, Sang-Jin;Shin, Woo-Shik;Chun, Gie-Taek;Jeong, Yong-Seob
    • KSBB Journal
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    • v.22 no.5
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    • pp.288-296
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    • 2007
  • Monascus pilosus (KCCM 60160) in submerged culture was optimized based on culture medium and fermentation conditions. Monacolin-K (Iovastatin), one of the cholesterol lowing-agent which was produced by Monascus pilosus may maintain a healthy lipid level by inhibiting the biosynthesis of cholesterol. Plackett-Burman design and response surface method were employed to study the culture medium for the desirable monacolin-K production. As a result of experimental designs, optimized production medium components and concentrations (g/L) were determined on soluble starch 96, malt extract 44.5, beef extract 30.23, yeast extract 15, $(NH_4)_2SO_4$ 4.03, $Na_2HPO_4{\cdot}12H_2O$ 0.5, L-Histidine 3.0, $KHSO_4$ 1.0, respectively. Monacolin-K production was improved about 3 times in comparison with shake flask fermentation of the basic production medium. The effect of agitation speed (300, 350, 400 and 450 rpm) on the monacolin-K production were also observed in a batch fermenter. Maximum monacolin-K production with the basic production medium was 68 mg/L when agitation speed was 500 rpm. And it was found that all spherical pellets (average diameter of $1.0{\sim}1.5mm$) were dominant during fermentation. Based on the results, the maximum production of 185 mg/L of monacolin-K with the optimized production medium was obtained at pH (controlled) 6.5, agitation rate 400 rpm, aeration rate 1 vvm, and inoculum size 3%.

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.

Production of Poly-3-hydroxybutyrate from Xylose by Bacillus megaterium J-65 (Bacillus megaterium J-65에 의한 xylose로부터 poly-3-hydroxybutyrate 생산)

  • Jun, Hong-Ki;Jin, Young-Hi;Kim, Hae-Nam;Kim, Yun-Tae;Kim, Sam-Woong;Baik, Hyung-Suk
    • Journal of Life Science
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    • v.18 no.12
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    • pp.1625-1630
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    • 2008
  • A microorganism capable of producing high level of poly-3-hydoxybutyrate (PHB) from xylose was isolated from soil. The isolated strain J-65 was identified as Bacillus megaterium based on the morphological, biochemical and molecular biological characteristics. The optimum temperature and pH for the growth of B. megaterium J-65 were $37^{\circ}C$ and 8.0, respectively. The optimum medium composition for the cell growth was 2% xylose, 0.25% $(NH_4)_2SO_4$, 0.3% $Na_2HPO_4{\cdot}12H_2O$, and 0.1% $KH_2PO_4$. The optimum condition for PHB accumulation was same to the optimum condition for cell growth. Copolymer of ${\beta}$-hydroxybutyric and ${\beta}$-hydroxyvaleric acid was produced when propionic acid was added to shake flasks containing 20 g/l of xylose. Fermenter culture was carried out to produce the high concentration of PHB. In batch culture, cell mass was 9.82 g/l and PHB content was 35% of dry cell weight. PHB produced by B. megaterium J-65 was identified as homopolymer of 3-hydoxybutyric acid by GC and NMR.

A CLINICAL STUDY OF THE JUDGMENT OF SASANG CONSTITUTION ACCORDING TO QUESTIONNAIRE (설문지(設問紙)를 통한 사상체질(四象體質)의 임상적(臨床的) 분류방안(分類方案) 연구(硏究))

  • Kim, Young-woo;Kim, Jong-weon
    • Journal of Sasang Constitutional Medicine
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    • v.10 no.1
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    • pp.215-233
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    • 1998
  • The object of this study is 27 patients who had been treated in the Oriental Medical Hospital at Dong Eui Medical Center during 6 months from January 1998 to June 1998. We proceeded the judgment of Sasang Constitution by Questionaire of Sasang Constitution Classification II(QSCC II). The following conclusion were made in comparison with Questionaire of Sasang Constitution Classification II(QSCC II) and Questionaire of Pusan Sasang seminar. 1. The subject of "the facial type is small and sharp" is significant differences in Sasang Constitution classification. The frequency of Soeumin group is more than Taeumin group and Soyangin group. 2. The subject of "the walking form is fast and shake the body" is significant differences in Sasang Constitution classification. The frequency of Taeumin group and Soeumin group is less than Soyangin group. 3. The subject of "the skin type is white and thin" is significant differences in Sasang Constitution classification. The frequency of Soeumin group is more than Soyangin group, and the frequency of Taeumin group is low marks. 4. The subject of "the skin type is tender and dry" is significant differences in Sasang Constitution classification. The frequency of Taeumin group and Soyangin group is less than Soeumin group. 5. The subject of "the image of face is smart" is significant differences in Sasang Constitution classification. The frequency of Soeumin group is more than Taeumin group and the frequency of Soyangin group is low marks. 6. The subject of "the sweating type is not sweatier" is significant differences in Sasang Constitution classification. The frequency of Taeumin group and Soeumin group is less than Soyangin group. 7. The subject of "the desire of eating is changeable accoding to emotion" is significant differences in Sasang Constitution classification. The frequency of Soeumin group is more than Taeumin group and Soyangin group. 8. The subject of "the health is changeable accoding to the type of stool" is significant differences in Sasang Constitution classification. The frequency of Soeumin group is more than Soyangin group and the frequency of Taeumin group is low marks. 9. The subject of "the type of voiding is changeable accoding to the drinking when they have a fever" is significant differences in Sasang Constitution classification. The frequency of Taeumin group and Soyangin group is less than Soeumin group. 10. The subject of "the skin type is soft" is significant differences in Sasang Constitution classification. The frequency of Soeumin group is more than Taeumin group and Soyangin group. 11. The subject of "the chief complaint is the forgetfulness" is significant differences in Sasang Constitution classification. The frequency of Soeumin group is less than Taeumin group and Soyangin group.

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Effects Of Environmental Factors And Individual Traits On Work Stress And Ethical Decision Making (간호사의 환경적 요소와 개인적 특성이 직무스트레스와 윤리적 의사결정에 미치는 영향)

  • Kim, Sang Mi L.;Shake ketefian
    • Journal of Korean Academy of Nursing
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    • v.23 no.3
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    • pp.417-430
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    • 1993
  • 이 연구는 환경적 요소(간호사의 자율성, 조직의 표준화)와 개인의 특성(통제위, 나이, 경험. 간호역할개념, 도덕성), 직무 스트레스, 윤리적 의사결정 사이의 관계를 이론적 틀을 구성하여 테스트함으로써 그 인과관계를 탐구하였다. 본 연구를 위해 개발된 모형은 1) Katz와 Kahn의 조직에 대한 개방체계 이론(open systems theory of organization) ; 2) Kahn. Wolfe, Quinn, Snoek의 스트레스 이론 (theory of stress) : 3) Kohlberg의 도덕발달 이론(theory of moral develop-ment): 그리고 4) 여러 문헌고찰을 기초로 하였다. 본 연구의 모형은 2가지의 주요 종속변수(직무 스트레스, 윤리적 간호행위), 2가지 매개변수(간호 역할개념, 도덕성 발달정도) 그리고 여러 독립변수들(조직의 표준화, 자율성, 통제위, 교육, 나이, 경험 등)로 구성되었다. 간단히 말해, 간호사의 스트레스와 윤리적 간호행위 를 개인 자신과 환경이라는 두 요소의 결과로 간주한 것이다. 미국(2개주)의 여러 건강관리기관에 근무하는 224명의 정규 간호사를 대상으로 하였고. 가설 검증을 위하여 1) 변수간의 인과관계를 조사하기 위한 Linear Structural Relationships(LISREL)기법과 2) 나이, 경험, 교육이 변수간의 관계에 미치는 중간역할을 알아보기 위해 상관분석을 이용하였다. LISREL결과를 보면 제시된 모델이 각 내재 변수에 상당한 설명력을 가지면서 자료에 잘 맞는 것으로 나타났다. 이 연구에서 가장 뚜렷한 점으로 나타난 것은 개인의 특성보다 환경적 요소로서의 자율성이 직무스트레스와 윤리적 의사결정을 예견하는데 훨씬 중요한 변수로 부각되었다는 점이다. 또한 간호사의 전문적 역할개념과 봉사적 역할개념이 간호사의 윤리적 의사결정을 예견하는 가장 중요한 요소로 나타났다. 중간영향(moderation effect)을 보면, 젊고 경험이 적은 간호사일수록 나이가 많고 경험있는 간호사보다 환경적 요소(자율성)에 더 큰 영향을 받는다는 것을 암시하고 있다. 또한 4년제 대학 이상을 졸업한 간호사의 윤리 적 간호행 위 는 2, 3년제 를 졸업 한 간호사 보다 환경적 요소에 의해 덜 영향을 받는 것으로 나타났다. 한편 자율성의 부족은 2, 3년제 졸업 간호사보다 4년제 졸업 간호사에게 더 심한 스트레스가 되고 있음을 시사하였다. 이 연구의 결과로부터 적어도 다음과 같은 두 가지 실제적인 제언을 도출할 수 있다. 첫째, 이 연구는 환경적요소로서의 자율성이 다른 어떤 개인적인 요소보다 직무 스트레스를 예견하는 데 중요한 요소라는 것을 제시하였다. 이것은 간호행정가들에게, 간호사의 직무 스트레스를 감소시키기 위해선 “자율성”이 아주 중요히 다루어져야 한다는 것을 의미한다. 만일 간호사들의 직무스트레스가 그 개인의 복지에 큰 해가 되고 환자를 간호하는 데 직접적으로 관계된다면, 간호행정가는 그 조직의 직무체계를 다시 평가해서 일에 대한 새로운 설계가 필요한지를 파악해야 한다. 또한 이 연구는 직무를 다시 설계할 경우, 누구에게 먼저 촛점을 두고 시작해야 하는지를 밝혀주고 있다. 즉, 젊고 경험이 미숙한 간호사들에게 촛점을 두고 시작해야 하며, 작업환경의 가장 중요한 차원중의 하나인 사회적 지원(social support)을 조심스럽게 고려해 보아야 한다. 둘째, 간호사의 윤리적 간호행위를 높히기 위해 전문적 역할개념과 봉사적 역할개념이 재강조될 필요가 있다. 이 두 역할개념 들을 교육을 통하여 효과적으로 가르칠 필요가 있다고 본다. 이 두 개념들이 간호사의 바람직한 간호행 위에 영향을 미치는 가장 중요한 요소로 나타났기 때문이다. 또한, 본 연구결과에 따르면, 경험이 많을수록 일에 싫증을 느껴 바람직한 윤리적 간호행위가 감소되는 경향이 있었다. 따라서, 건강관리체제 (health care system) 안에서의 간호사의 역할이-전문직으로서의, 그리고 환자를 위한 옹호자로서의-학교와 임상에서 효과적으로 교육되어져야 한다고 본다. 간호사들의 역할에 대한 계속적인 교육이 학생은 물론 임상 간호사들에게도 실시되어져야 할 것이다. 미래연구의 방향을 제시해 보면 첫째로 연구의 일반화를 높히기 위해 더 많은 대상자를 포함시켜야 한다. 이는 여러 종류의 표본을 반드시 한번에 전부 포함시켜야 한다는 것을 의미하는 것이 아니고, 특정한 여러 표본들을 연속적으로 연구함으로서 이 목표를 성취할 수 있다고 생각한다. 둘째는 여러 construct들(윤리적 간호행위, 직무 스트레스, 간호 역할개념 등)에 대한 적절한 측정도구를 개발해야 한다. 측정도구를 개발하기 위해서는 풍부하고 세세한 통찰력을 제공하는 질적인 정보를 얻는 것이 선행되어야 한다. 셋째, 윤리적 간호행위와 직무 스트레스에 관한 연구를 증진시키기 위해 실험설계 및 종단적 연구(expel-imental, longitudinal design)가 시도될 필요가 있다. 마지막으로, 윤리적 간호행위와 직무 스트레스를 예견할 수 있는 이론적 탐구(theoretical exploration), 즉 이론정립을 위하여, 환경적 요소와 개인의 특성에 대한 자세한 정보를 제공해 줄 수 있는 질적 연구들이 요구된다.

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Enhanced production of monacolin-K through supplement of monacolin-K precursors into production medium and cloning of SAM synthetase gene (metK) (Precursor제공 및 생합성 관련 유전자의 cloning을 통한 Monacolin-K 생산성 향상)

  • Lee, Mi-Jin;Jeong, Yong-Seob;Chun, Gie-Taek
    • KSBB Journal
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    • v.23 no.6
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    • pp.519-524
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    • 2008
  • Monacolin-K is a strong anti-hypercholesterolemic agent produced by Monascus sp. via polyketide pathway. High-yielding mutants of monacolin-K were developed through rational screening strategies adopted based on understanding of monacolin-K biosynthetic pathway. Through the experiments for investigating various amino acids as putative precursors for the monacolin-K biosynthesis, it was found that production level of monacolin-K was remarkably increased when optimum amount of cysteine was supplemented into the production medium. We suggested that these phenomena might be related to the special roles of SAM (S-adenosyl methionine), a putative methyl group donor in the biosynthetic pathway of monacolin-K, demonstrating close interrelationship between SAM-synthesizing primary metabolism and monacolin-K synthesizing secondary metabolism. Namely, increase in the intracellular amount of SAM derived from the putative precursor, cysteine which was extracellularly supplemented into the production medium might contribute to the significant enhancement in the monacolin-K biosynthetic capability of the highly mutated producers. On the basis of these assumptions derived from the above fermentation results, we decided to construct efficient expression vectors harboring SAM synthetase gene (metK) cloned from A. nidulans, with the hope that increased intracellular level of SAM could lead to further enhancement in the monacolin-K production through overcoming a rate-limiting step associated with monacolin-K biosynthesis. Hence, in order to overcome the plausible rate-limiting step associated with monacolin-K biosynthesis by increasing intracellular level of SAM, we transformed the producer mutants with an efficient expression vector harboring gpdA promoter of the producer microorganism, and metK gene. Notably, from the resulting various transformants, we were able to screen a very high-yielding transformant which showed approximately 3.3 fold higher monacolin-K productivity than the parallel nontransformed mutants in shake flask cultures performed under the identical fermentation conditions.

A Comparative Study of the Oriental and the Occidental Medical Literature on the Etiologies & Treatments for Palmoplantar Hyperhidrosis (수족한(手足汗)의 원인(原因)과 치료법(治療法)에 대한 동(東).서의학적(西醫學的) 고찰(考察))

  • Ko, Young-Chul;Shin, Jo-Young
    • The Journal of Internal Korean Medicine
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    • v.18 no.2
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    • pp.268-295
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    • 1997
  • Sweating is natural phenomenon necessary for the regulation of an individual's body-temperature. The secretion of sweat is mediated by a portion of our vegetative nervous system(the sympathetic nervous system). In some people, this system is working at a very high activity level, far higher than needed to keep a constant temperature. This condition is referred to as hyperhidrosis. Especially excessive sweating of the hands and the feet is palmoplantar hyperhidrosis or volar hyperhidrosis. This is by far the most distressing condition. It was founded that the first important cause of this was emotional factor. The hands are much more exposed in social and prefessional activities than any other part of our body. Many individuals with palmoplantar hyperhidrosis are limited in their choice of proffession, because unable to manipulate materials sensitive to humidity or reluctant to shake hands; some patients arrive to the point to avoid social contact. The occidental medical treatments for palmoplantar hyperhidrosis include application of topical agents(chemical antiperspirants such as aluminum chloride), iontophoresis(treatment with electrical current), or surgery(thoracic sympathectomy). It was reported that the most effective treatment was thoracic sympathectomy. So this study was started to find the easy and effective oriental medical treatments against the occidental medical treatments through the oriental medical literature. The occidental medical idea for palmoplantar hyperhidrosis is only limited in neurologic system, so surgery is the best treatment. But the oriental medical idea for palmoplantar hyperhidrosis is much wider, so the oriental medical causes and treatments for this are able to be veriety. And the oriental medical teatment is freely in treating the patients of palmoplantar hyperhidrosis, because entire idea including pulse, facial color, mental condition, constitution and other symptom exists in the oriental medicine. The results of a bibliographic study of causes and treatments for palmoplantar are as follows; 1. The main causes of pa1moplantar hyperhidrosis are heat in the stomach, damp-heat in the spleen and the stomach, insufficiency of the spleen-qi and the stomach-qi, deficiency of the spleen-yin and the stomach-yin, and the others are the stomach-cold syndrome, stasis of blood and dyspepsia in the stomach, disorder of the liver-qi, deficiency of the heart-yin and the kidney-yin, deficiency of the heart-yang and the kidney-yang, stagnated heat in the liver and the spleen, the lung channel-heat etc. 2. The main methods of medical treatments for palmoplantar hyperhidrosis are clearing out the stomach-heat, eliminating dampness and heat in the spleen and the stomach, invigorating the spleen-qi and the stomach-qi, reinforcing the spleen-yin and the stomach-yin, warming the stomach, relaxing the liver and alleviating of mental depression and tonifying the heart and the kidney etc. 3. The main prescriptions of palmoplantar hyperhidrosis are Taesihotang, Palmultang-kakam, Samyeongbaechusan, Chongbisan, Sasammaekmundongtang, the others are Leejungtang, Hwangkikonjungtang, Seungkitang, Boyumtang, Baekhotang, Chongsimyonjayum, Moyrosan, etc. 4. Local medicine for external use are liquid after boiling alum in water for about 1 or 2 hours, liquid after boiling alum and pueraria root in water and liquid after boiling stragalus root, pueraria root, ledebouriella root and schizonepeta in water, etc. 5. The methods of acupuncture therapy include invigorating Bokyru, Yumkuk and purgating Hapkouk, or invigorating Bokyru, Kihae and purgating Hapkouk, or steadying Hapkouk, Nokung.

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Optimization of Large Scale Culture Conditions of Bacillus ehimensis YJ-37 Antagonistic to Vegetables Damping-off Fungi (채소류 모잘록병균에 길항하는 Bacillus ehimensis YJ-37의 대량배양 최적조건)

  • 주길재;김진호
    • Journal of Life Science
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    • v.12 no.3
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    • pp.242-249
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    • 2002
  • The optimal culture conditions in 500$m\ell$ flask suture, 5$\ell$ jar fermenter and 2,000 $\ell$ large stale culture were investigated to maximize the production of antibiotic on Rhizoctonia solani AC4, the causal agent of vegetables damping-off, by the strain Bacillus ehimensis YJ-37. Starch (1.5%) as a carbon source, peptone (0.6%) as a nitrogen source and MgC1$_2$(0.15%) as a metal ion in the medium containing N $a_2$HP $O_4$(0.3%) showed the highest production of the antibiotic(s) in a rotary shake (200 rpm). Optimal initial pH of the culture medium, culture temperature and culture time for the antibiotic(s) production were pH 8.0, 32$^{\circ}C$ and 54hrs, respertively. Under the optimal renditions in flask culture, cell growth and antifungal activity (clear zone size) were 1.42 $\times$ 10$^{8}$ cfu/$m\ell$ (21g-DCW/ $\ell$) and 13.9 mm, respectively. In 5 $\ell$ jar fermenter (medium 3 $\ell$), optimal air flow, agitation speed and culture time for the antibiotic(s) production were 2 vvm, 200 rpm and 48hrs, respectively. Under the optimal conditions in 5 $\ell$ jar fermenter, tell growth and antifungal activity were 2.06 $\times$ 10$^{8}$ cfu/$m\ell$ (30g-DCW/ $\ell$) and 13.4 mm, respectively. Under the culture conditions of air flow (30 vvm) and agitation speed (200 rpm) at 32$^{\circ}C$ for 10 days in 2,000 $\ell$ large scale culture (medium 1,800 $\ell$, pH 8.0), cell growth and antifungal activity were 0.81$\times$10$^{8}$ cfu/$m\ell$ (12g-DCW/ $\ell$) and 8.6 mm, respectively.

Development of a Simultaneous Analysis Method for DDT (DDD & DDE) in Ginseng (인삼 중 DDT(DDD 및 DDE) 분석법의 개발)

  • Kim, Sung-Dan;Cho, Tae-Hee;Han, Eun-Jung;Park, Seoung-Gyu;Han, Chang-Ho;Jo, Han-Bin;Choi, Byung-Hyun
    • Korean Journal of Food Science and Technology
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    • v.40 no.2
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    • pp.123-128
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    • 2008
  • The MRLs (maximum residue limits) of DDT (DDD and DDE) in fresh ginseng, dried ginseng, and steamed red ginseng are set as low as 0.01 mg/kg, 0.05 mg/kg, and 0.05 mg/kg, respectively. Therefore, this study was undertaken to develop a simple and highly sensitive analysis method, as well as to reduce interfering ginseng matrix peaks, for the determination of DDT isomers (o,p'-DDE, p,p'-DDE, o,p'-DDD, p,p'-DDD, o,p'-DDT, and p,p'-DDT) in fresh ginseng, dried ginseng, and steamed red ginseng at the 0.01 mg/kg level. The method used acetonitrile extraction according to simultaneous analysis, followed by normal-phase Florisil solid-phase extraction column clean-up. The purification method entailed the following steps: (1) dissolve the concentrated sample extract in 7 mL hexane; (2) add 3 mL of $H_2SO_4$; (3) vigorously shake on avortex mixer; (4) cetrifuge at 2000 rpm for 5 min; (5) transfer 3.5 mL of the supernatant to the Florisil-SPE (500 mg/6 mL);and (6) elute the SPE column with 1.5 mL of hexane and 10 mL of ether/hexane (6:94). The determination of DDT isomers was carried out by a gas chromatography-electron capture detector (GC-${\mu}$ECD). The hexane and ether/hexane (6:94) eluate significantly removed chromatographic interferences, and the addition of 30% $H_2SO_4$ to the acetonitrile extract effectively reduced many interfering ginseng matrix peaks, to allow for the determination of the DDT isomers at the 0.01 mg/kg level. The recoveries of the 6 fortified (most at 0.01 mg/kg) DDT isomers from fresh ginseng, dried ginseng, and steamed red ginseng ranged from 87.9 to 99.6%. The MDLs (method detection limits) ranged from 0.003 to 0.009 mg/kg. Finally, the application of this method for the determination of DDT isomers is sensitive, rapid, simple, and inexpensive.