• 한국생물공학회:학술대회논문집
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• 2002.04a
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• pp.461-464
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• 2002

A Chemoautotroph identified as an Aeromonas sp. strain JS-1 was isolated from fresh water. Aeromonas sp. strain JS-1 used the $H_2$ and $CO_2$ as energy and carbon sources, respectively. Growth characteristics for improving the $CO_2$ fixation rate were examined in batch cultivation. Its results shown that the optimal growth appeared at culture conditions of $35^{\circ}C$, pH 7 and NaCl 0.1%(w/v). Some hydrogen-oxidizing bacteria were reported that the enzyme activity of ribulose 1,5-bisphosphate carboxylase- oxygenase (RubisCO-EC 4.1.1.39), in the key enzyme of the Calvin-Benson cycle. A RubisCO was purified from a chemoautotrophic bacterium, Aeromonas sp. strain JS-1. the enzyme was purified by ammonium sulfate precipitation, DEAE-sepharose CL-6B and gel filtration chromatography. The RubisCO showed that molecular mass was about 560KDa from gel filtration chromatography and nondenaturing PAGE, and the RubisCO was confirmed to consist of $L_8S_8$ enzyme structure by sodium dodecyl sulfate polyacrylamide gel electrophoresis. A large subunit was about 56KDa and small one was about 15kDa. The Km values of the enzyme for ribulose 1,5-bisphosphate(RUBP), $NaH^{14}CO_3$, and $Mg^{++}$ were estimated to be 0.25mM, 5.2mM, and 0.91mM, respectively. The optimum temperature for RubisCO enzymatic activity were $50^{\circ}C$, and the enzymatic activity was stable up to $45^{\circ}C$.

• Journal of Microbiology and Biotechnology
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• v.28 no.10
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• pp.1749-1759
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• 2018

Recombinant (rec) prion protein (PrP) is an extremely useful resource for studying protein misfolding and subsequent protein aggregation events. Here, we report mass production of high-purity rec-polypeptide encoding the C-terminal globular domain of PrP; (90-230) for human and (89-231) for murine PrP. These proteins were expressed as His-tagged fusion proteins in E. coli cultured by a high cell-density aerobic fermentation method. RecPrPs recovered from inclusion bodies were slowly refolded under reducing conditions. Purification was performed by a sequence of metal-affinity, cation-exchange, and reverse-phase chromatography. The current procedure yielded several dozens of milligrams of recPrP per liter with >95% purity. The purified recPrPs predominantly adopted an ${\alpha}$-helix-rich conformation and were functionally sufficient as substrates to measure the seeding activity of human and animal prions. Establishment of a procedure for high-level production of high-purity recPrP supports the advancement of in vitro investigations of PrP including diagnosis for prion diseases.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2019.04a
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• pp.56-56
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• 2019

본 연구는 식용과 약용으로 이용되는 국내 자생식물인 야산고비(Onoclea sensibilis var. interrupta Maxim.)를 기내에서 대량증식하기 위한 조건을 구명하기 위하여 수행되었다. 무가온 온실에서 수집한 포자를 기내에서 발아시켜 전엽체를 획득하였으며, 계대배양하며 실험의 재료로 사용하였다. 전엽체의 대량증식을 위해 전엽체 0.3g을 메스로 균일하게 다진 후 증류수 1ml와 함께 배지에 고루 펼쳐서 배양하는 방법을 사용하였으며, 증식에 미치는 배지의 영향을 확인하기 위하여 1/4, 1/2, 1, 2배로 조절한 MS배지를 조성하여 8주간 배양하였다. 이후, 증식이 우수한 배지를 선정하여 sucrose와 질소급원의 농도를 조절하였으며, 활성탄을 첨가하여 증식에 미치는 영향을 확인하였다. 배지종류 실험의 결과, 생체중이 1MS에서 10.2g으로 초기 접종량에 비해 가장 많이 증가하였으며, 현미경을 이용하여 관찰한 결과에서도 정상적인 전엽체의 형태인 심장형으로 발달하였다. 증식이 우수하였던 1MS배지에 sucrose의 농도를 조절하여 배양한 결과에서는 1%의 처리구에서 가장 증식률이 좋았으며, 질소급원의 경우 30mM의 농도로 조절한 처리구가 가장 좋은 결과를 보였다. 배지 내 활성탄의 첨가는 처리구당 증가된 전엽체의 생체중이 유의적인 차이를 보이지 않았다. 포자체 대량 형성을 위한 적정 배양토의 혼합조건을 확인하기 위하여 원예상토, 피트모스, 펄라이트 및 마사토의 혼합비율을 5종류로 달리하여 조성하여 사각분($7.5{\times}7.5{\times}7.5cm$)에 충진하였다. 조성한 배양토에 기내에서 배양한 전엽체 1g을 증류수와 함께 블렌더를 이용하여 10초간 분쇄하여 토양표면에 고루 분주하였다. 이후 12주간 재배한 결과, 모든 토양조합에서 포자체가 형성되었다. 그중 원예상토와 마사토를 2:1(v:v)로 혼합한 토양에서 포트 당 405.0개로 가장 많은 포자체가 형성되었으며, 전체적인 생육 또한 비교적 양호한 결과를 보였다. 따라서 야산 고비의 전엽체 대량증식에 적합한 배지는 경제성과 생육수준을 고려하여 1%의 sucrose와 질소급원의 농도를 30mM로 조절한 1MS 배지가 적합하며, 포자체 대량생산을 위해서는 원예상토, 마사토를 2:1(v:v)로 혼합한 토양이 적합하다고 판단되었다.

• Korean Journal of Plant Resources
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• v.32 no.4
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• pp.290-295
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• 2019

This study was carried out to select the optimal condition for prothallus propagation and sporophyte formation of Leptogramma pozoi (Lag.) Ching subsp. mollissima (Fisch. ex Kunze) Nakaike and to provide basic data for mass production system. In the propagation, 300 mg of prothallus were inoculated in different kinds of medium [1/4, 1/2, Murashige and Skoog (MS), Knop], sucrose (0, 1, 2, 3, 4%), and cultured for 8 weeks. Sporophyte formation studies were carried out by inoculating blended prothallus into artificial soils. The soils were mixed with horticultural substrate, peatmoss, perlite, and decomposed granite at different ratios or only with the horticultural substrate. Then the prothallus was cultivated for 12 weeks to figure out the formation and growth of the sporophytes. The growth and development of prothalli were excellent on MS medium. According to medium components, prothallus growth was favorable in all treatments except 0% sucrose treatment and the highest in 2% sucrose. In the experiment of soil mixtures, sporophyte formation was the highest in the horticultural substrate:perlite 2:1 (v:v) and horticultural substrate:decomposed granite 2:1 (v:v) treatment, and the overall growth was good in the horticultural substrate:decomposed granite 2:1 (v:v) mixed soil.

• Journal of Microbiology and Biotechnology
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• v.32 no.5
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• pp.630-637
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• 2022

The objective of this study was to optimize industrial-grade media for improving the biomass production of Weissella cibaria JW15 (JW15) using a statistical approach. Eleven variables comprising three carbon sources (glucose, fructose, and sucrose), three nitrogen sources (protease peptone, yeast extract, and soy peptone), and five mineral sources (K₂HPO₄, potassium citrate, ⳑ-cysteine phosphate, MgSO₄, and MnSO₄) were screened by using the Plackett-Burman design. Consequently, glucose, sucrose, and soy peptone were used as significant variables in response surface methodology (RSM). The composition of the optimal medium (OM) was 22.35 g/l glucose, 15.57 g/l sucrose, and 10.05 g/l soy peptone, 2.0 g/l K₂HPO₄, 5.0 g/l sodium acetate, 0.1 g/l MgSO₄·7H₂O, 0.05 g/l MnSO₄·H₂O, and 1.0 g/l Tween 80. The OM significantly improved the biomass production of JW15 over an established commercial medium (MRS). After fermenting OM, the dry cell weight of JW15 was 4.89 g/l, which was comparable to the predicted value (4.77 g/l), and 1.67 times higher than that of the MRS medium (3.02 g/l). Correspondingly, JW15 showed a rapid and increased production of lactic and acetic acid in the OM. To perform a scale-up validation, batch fermentation was executed in a 5-l bioreactor at 37℃ with or without a pH control at 6.0 ± 0.1. The biomass production of JW15 significantly improved (1.98 times higher) under the pH control, and the cost of OM was reduced by two-thirds compared to that in the MRS medium. In conclusion, OM may be utilized for mass producing JW15 for industrial use.

• Journal of Plant Biotechnology
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• v.44 no.4
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• pp.484-489
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• 2017

Lilies as cut flowers are one of the most popular ornamental plants in South Korea. It is necessary to develop lily cultivars with high qualities. Therefore, highly efficient propagation systems are needed following release of elite cultivars. In this study, we used taurine treatment to improve the growth conditions including shoot and bulb formation, fresh weight gain, and reduction of rooting and browning. We experimentally evaluated the effect of taurine as a growth stimulator, at concentrations of 0, 2.5, 5, 10, 15 and 20 mg/l. The results showed that 20 mg of taurine enhanced shoot formation by 85% and increased fresh weight 5.5-fold, which was higher than the approximately four-fold increase in the control. In addition, multiple bulb formation rate was increased by 80% and rooting by 82% following exposure to 20 mg/l of taurine. The efficiency of taurine treatment was higher than that of control with 50% multiple bulb formation rate and 60% rooting rate. The browning was 10.6% at 2.5 mg/l of taurine when compared with 0.8% at 20 mg/l. Taurine showed a positive effect on the overall growth of lily plants in terms of increased fresh weight, shoot formation rate, rooting, and formation of multiple bulbs, indicating that taurine can be used as an alternative to amino acids or as an antioxidant such as citrate and vitamin C in plant tissue culture.

• Korean Journal of Environmental Biology
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• v.32 no.1
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• pp.58-67
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• 2014

The outdoor mass cultivation is not possible for microalgae in Korea all year round, due to cold winter season. It is not easy to maintain proper level of productivity of microalgae even in winter. To prevent a drastic decrease of temperature in a greenhouse, two layers were covered additionally, inside the original plastic layer of the greenhouse. The middle layer was made up of plastic and the inner layer, of non-woven fabric. Acrylic transparent bioreactors were constructed to get more sunlight, not only from the upper side but also from the lateral and bottom directions. In winter at freezing temperatures, six different culture conditions were compared in the triply covered, insulated greenhouse. Wastewater after anaerobic digestion was used for the cultivation of microalgae to minimize the production cost. Water temperature in the bioreactors remained above $10^{\circ}C$ on average, even without any external heating system, proving that the triple-layered greenhouse is effective in keeping heat. Algal biomass reached to 0.37g $L^{-1}$ with the highest temperature, in the experimental group of light-reflection board at the bottom, with nitrogen and phosphorus removal rate of 92% and 99%, respectively. When fatty acid composition was analyzed using gas-chromatography, linoleate (C18 : 3n3) occupied the highest proportion up to 61%, in the all experiment groups. Chemical oxygen demand (COD), however, did not decrease during the cultivation, but rather increased. Although the algal biomass productivity was not comparable to warm seasons, it was possible to maintain water temperature for algae cultivation even in the coldest season, at the minimum cost.

• Korean Journal of Environmental Biology
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• v.38 no.3
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• pp.404-415
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• 2020

We investigated the seasonal variations in growth and physiological responses of the kelp species Ecklonia cava to different nitrogen sources to establish indoor culture conditions for mass production. Ecklonia cava was cultivated for 10 days in 16 combinations of seawater temperatures (15, 17, 21, and 25℃) and different nitrogen sources (control; NH-NH₄⁺ 100 μM; NO-NO₃^- 100 μM; and NHNO-NH₄⁺ 50 μM+NO₃^- 50 μM). The growth and growth rate of the blade were affected by temperature. The mean fresh weight and area-based daily growth rate were the highest (5.8±0.5 and 6.6±0.5% day^-1, respectively) at 15℃ and the lowest (2.2±0.2 and 3.0±0.3% day^-1, respectively) at 25℃. The daily growth rate was the highest in the NH and NO treatments and lowest in the control. The nitrate reductase activity of E. cava varied with water temperature (season). The highest activity was in the control (1.32±0.10 μmol NO₂^- g^-1 dry weight h^-1) and the lowest was in the NH treatment (0.25±0.02 μmol NO₂^- g^-1 dry weight h^-1). The photosynthetic pigment concentrations reached a maximum value in the NHNO treatment and a minimum value in the control. These results showed that water temperature played an important role in the cultivation of E. cava and that a single supply of NH₄⁺ or NO₃^- may induce the accelerated growth of E. cava. The growth and physiological responses of E. cava to different nitrogen sources during each season provide valuable information for determining the optimal nitrogen source in E. cava cultivation under indoor conditions.