• KSBB Journal
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• v.11 no.2
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• pp.201-210
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• 1996

The production of polyhydroxyalkanoate(PHA) from glucose by batch and fed-batch culture of Pseudomonas sp. HJ was studied. In batch culture using fermentor, 400 rpm of agitalion speed, 2 vvm of aeration rate, 18 hours of inoculum age, and 5% (vlv) of inoculum size were optimal. PHA production was not increased by deficiency of oxygen. In a batch culture, the final call mass was $6.251g/\ell$, and PHA content was 20% of dry cell weight. In a constant feeding fed-batch culture, cell mass increased to $33.24g/\ell$, and PHA content reached 48.9% of dry cell weight. In an intermittent feeding fed-batch culture, cell mass increased to $37.89g/\ell$, and PHA content reached 53.5% of dty cell weight.

• Journal of fish pathology
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• v.10 no.2
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• pp.177-186
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• 1997

To establish effectively for isolation and cultivation and cultivation of scuticociliata, we tried to culture scuticociliata using culture medium(P2Y1-DW medium). The number of scuticociliata reached $1.50{\times}10^3$ cells/ml at the temperature of $25^{\circ}C$ in 3 days, but $1.03{\times}10^2$ cells/ml at $15^{\circ}C$. In P1Y1-S medium growth reached $2.6{\times}10^4$cells/ml in 3 days, but the number in DW medium was the lowest with $2.0{\times}10^2$cells/ml. In all these media, the number decreased after 5 days. As the method of the parasite`s division, after attaching the anterior part of the body, its shape gradualy changed to a western pear-like appearance. The parasite completely divided into two cells, then freely swam and became an adult cell, size of $40{\mu}m$ in size within about 3 hours from attaching.

• Microbiology and Biotechnology Letters
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• v.26 no.3
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• pp.200-205
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• 1998

A probiotic-strain of Lactobacillus sp. was cultured in bovine blood plasma-based (BBPB) medium and freeze-dried to prepare a probiotic product as an animal feed additive. The cell mass produced in the medium, $5.2{\times}10^9$ CFU/ml, was high enough to be commercialized and was 74% of that in MRS medium. The survival rate of tactobacillus sp. against freeze-drying was affected by the conditions for treatment of cultured BBPB broth before freeze-drying such as pH adjustment, volume reduction and freezing rate. It was also found that the blood protein hydrolysate remaining in broth also enhanced the survival rate. Among various protective substances, sucrose showed a high stabilizing effect with 10% (w/v) addition, by which the maximum survival rate (48.3%) and viable cell count ($3.0{\times}10^{10}$ CFU/g) were obtained.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.88-88
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• 2001

소 난포란을 체외에서 성숙, 수정시킨 후, 2~8 세포기 수정란을 체외배양액인 CR$_1$aa에 일정량의 nitric oxide scavenger, Hb와, inhibitor인 L-nitro - L- arginine methyl ester (L-NAME)를 첨가 배양하여 체외발육에 미치는 영향을 검토하고자 실시하였다. 체외수정 후 체외배양 48시간에 난구세포의 제거 유.무에 따라 CR$_1$aa 배양액에 hemoglobin을 0, 1, 5$\mu\textrm{g}$/$m\ell$를 첨가한 구의 상실배기이상 체외발육성적은 난구세포를 제거한 구에서 23.8%, 33.3% 및 26.8%였으며, 난구세포를 제거하지 않은 구에서는 각각 39.5%, 54.8% 및 48.8%로써 난구세포를 제거하지 않은 구에 Hb를 1$\mu\textrm{g}$/$m\ell$를 첨가한 구가 여타구 보다 통계적으로 높은 결과를 얻었다(P<0.05). 체외수정 후 체외배양 96시간 후 난구세포를 제거 유.무에 따라 Hb를 0, 1, 5$\mu\textrm{g}$/$m\ell$를 첨가하였을 때, 상실배기 이상 체외발육성적은 난구세포를 제거한 구에서 각각 28.6%, 46.2% 및 39.1%였으며, 난구세포를 제거하지 않은 구에서는 각각 33.9%, 67.2% 및 46.0%로써, 난구세포를 제거하지 않은 구에 Hb를 1$\mu\textrm{g}$/$m\ell$첨가구가 여타구에 비해 통계적으로 유의하게 높게 나타났다( P<0.05).CR$_1$aa배양액에 L-NAME를 0, 10, 50 및 100mM을 첨가한 구에서 상실배이상 발육된 체외발육성적은 각각 55.6%, 64.9%, 58.8% 및 66.7%로써 L-NAME 첨가구가 무첨가구에 비해 커다란 차이가 나타나지 않았다(P>0.05). 본 실험의 결과로 볼 때, nitric oxide scavenger인 hemoglobin의 첨가는 체외수정 후 체외배양 96시간에 1$\mu\textrm{g}$/$m\ell$ 첨가하는 것이 체외수정란의 체외 발육율을 향상시켰으며, nitric oxide inhibitor인 L-NAME는 첨가 농도에 관계없이 체외수정란의 체외발육율에 영향을 미치지 않는 것으로 나타났다. 본 실험에서 생산된 배반포기 체외수정란의 세포수에는 커다란 차이가 인정되지 않았다.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2010.10a
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• pp.19-19
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• 2010

황칠나무(Dendropanax morbifera Lev.)는 두릅나무과(Araliaceae)에 속하며 학명에서 뜻하는 바와 같이 목본 (Dendro), 전능약(Panax)이라는 의미가 있고 나무인삼이라 불리기도 하며 줄기에 상처를 내면 노란액이 나온다고 해서 황칠나무(D. morbifera)라는 이름이 붙여졌다. 두릅나무과는 우리나라에서 최고의 약재들로 손꼽히는 인삼(Panax ginseng), 가시오갈피(Eleutherococcus senticosus) 등의 약용식물을 포함하고 있어서 황칠나무는 황칠수지액 이외에 약용식물로서의 무한한 개발 가능성을 내포하고 있다. 따라서 본 실험은 황칠나무의 기내 부정근 유도 및 증식조건의 확립을 목적으로 수행되었다. 우선 황칠나무의 기내 발아체로부터 부위(잎, 줄기, 뿌리)를 달리하여 부정근을 유도한 결과, 잎은 줄기나 뿌리보다 양호한 부정근의 유도를 보였다. 또한 유도된 부정근을 이용하여 옥신의 종류에 따른 부정근 유도율을 조사한 결과 IBA와 NAA는 IAA와 2.4-D보다 높은 유도율을 보였다. IBA의 농도에 따른 유도율과 증식효율은 IBA가 1.0 mg/L 첨가되었을 때 가장 높은 유도 및 증식효율을 보였다. 최적의 액체배지조건을 확인하고자 sucrose의 농도와 염농도를 달리하여 실험한 결과 1/2MS 배지는 MS 배지보다 10%정도 높은 증식율을 보였다. 액체배양 된 황칠나무의 부정근을 각각 1/2MS 배지에 30 g/L sucrose, 3.0 mg/L IBA가 첨가된 5 L volume 생물반응기에 4주 간 배양한 대조구와 2주 후 IBA의 농도를 1.0으로 낮추어 배양한 실험구에서 2주후 IBA의 농도를 낮추어 배양한 실험구에서 대조구보다 약 2배 높은 부정근의 증식량을 보였다. 결국, 황칠나무의 종자발아체를 이용하여 부정근의 유도 및 증식조건에 필요한 기내배양조건을 확립하였고, 플라스크와 생물반응기 배양을 통해 효율적인 실험실 내 증식조건을 확립하였다. 본 실험결과는 향후 황칠나무 천연추출물을 활용한 향장품/식,의약품 소재의 대량확보 차원에서 중요한 가치를 내포하고 있다고 조심스럽게 사료된다.

• KSBB Journal
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• v.15 no.1
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• pp.35-41
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• 2000

We tried to prepare encapsulated whole cell cyclodextrin glucanotransferase(CGTase) in order to produce glycosyl-xylitol using xylitol as glucosyl acceptor. The organic nitrogen source was more effective for the production of CGTase from Bacillus macerans IFO 3490 than the inorganic one. Most of the CGTase which had been produced during cultivation was excreted to the growth medium. B. macerans cells inocculated in the capsule failed to grow to the high cell density. Adsorbents such as activated charcoal, Sephadex and Amberite resins could not adsorb efficiently the CGTase from the broth solution. We obtained successfully the encapsulated whole cell CGTase by immobilizing the concentrated broth solution in the calcium alginate capsules. The encapsulated whole cell CGTase carried out the transglycosylation reaction which converts xylitol into glucosyl-xylitol using dextrin as glucosyl donor.

• Microbiology and Biotechnology Letters
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• v.16 no.6
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• pp.450-456
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• 1988

For optimal production of L-tryptophan using a regulatory mutant of Escherichia coli the relationship between product formation and acid production was investigated. Experimental results showed that the production level of L-tryptophan was lowered as the specific acid production rate increased. In order to reduce the amount of acid produced during the fermentation, a controlled fed-batch fermentation was employed. In this fed-batch process, the feed rate of the nutrient feed medium was controlled in relation to the oxygen level in the culture and thus the growth of the cells was regulated in such n way that the oxygen demand of the culture could not exceed the oxygen sup-ply. When E. coli cells were cultivated in a controlled fed-batch mode of tormentor operation, the specific acid production rate was significantly reduced and L-tryptophan production was increased as much as five times that obtained in a conventional fed-batch fermentation.

• Journal of Plant Biotechnology
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• v.42 no.3
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• pp.239-244
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• 2015

Calanthe striata f. sieboldii Decne. ex Regel is a terrestrial orchid with beautiful flowers arranged in racemose inflorescences. This species is threatened due to over-collection and loss of suitable habitats. Asymbiotic germination is useful in the conservation efforts to re-establish plants in the wild, and for commercial propagation. In this study, we investigate the effects of sodium hypochlorite (NaOCl), three types of culture media (Phytomax Orchid Maintenance - POM, Seed Germination Maintenance - SGM, and Murashige and Skoog 1962 - MS), and plant growth regulators on embryo swelling, protocorm formation, and embryo diameter of C. striata f. sieboldii. Treatment with 1% NaOCl for 30 min greatly enhanced embryo swelling (28.3%), embryo diameter ($205.8{\mu}m$), and embryo protocorm formation (54.8%) compared to seeds without NaOCl treatment (embryo swelling 8.5%, embryo diameter $14.6{\mu}m$, and protocorm formation 13.4%) on POM medium. Protocorm formation on POM medium supplemented with 1.0 mg/L N6-benzyladenine (BA) (95.6%) was better than the control (54.5%). Additionally, the effects of activated charcoal (AC) and sucrose on seedling growth in in vitro culture were examined. The protocorm converted into healthy plants with well-developed shoot primordia on the POM medium with AC and sucrose. The most suitable conditions for seedling growth after 10 weeks of culture were the POM medium with AC or sucrose. These results show effective asymbiotic seed germination and in vitro seedling growth of C. striata f. sieboldii.

• Journal of Plant Biotechnology
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• v.39 no.3
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• pp.163-168
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• 2012

An optimization in vitro seed germination was established by using triphenol tetrazolium chloride (TTC) test, which has been known as two rare orchids (Gastrodia verrucosa Blume and Hetaeria sikokiana Tuyama) in Jeju island. We have established proper NaOCl treatment for in vitro germination of G. verrucosa and H. sikokiana through TTC test. In the case of H. sikokiana, seed viability through TTC test was high with 95% in control. However, NaOCl 1% treatment for 30 minutes showed the highest embryo swelling rate to seed viability. Likewise, swelling formation of embryos, diameter of embryos, protocorm formation and diameter of protocorms of G. verrucosa was 87%, $59{\mu}m$, 91% and $138{\mu}m$ through NaOCl 1% treatment for 30 minutes. This result will be applied on the basic information for improving in vitro seed germination rate of G. verrucosa and H. sikokiana.

• Korean Journal of Agricultural Science
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• v.22 no.1
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• pp.62-68
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• 1995

This study was carried out to examine splitting, developmental capacity and rapid freezing of blastomeres separated from 2-, 4-, 8-cell and morula from porcine embryos. The results obtained in this study were summerized as follows : 1. The successful splitting rate by pronase was 85.7% in 2-cell embryos(average splitting rate, 68.0%), and by manipulator was 76.6% and 74.3% in 2- and 4-cell embryos. 2. The developmental capacity rates of splitted embryos by the pronase treatment were 24.1%, 20.4%. 25.5% and 26.6% in 2-, 4-, 8-cell and morula, and by manipulator were 36.4%, 39.5%, 36.1% and 41.9%, respectively. 3. The successful results of in vitro culture after frozen-thawed of splitted embryos were 16.1%(glycerol) in 2-cell, 16.7%(DMSO) in 4-cell and 27.6%(ethyleneglycol) in morula, respectively.