• 한국미생물·생명공학회지
• /
• 제26권2호
• /
• pp.130-136
• /
• 1998

참당귀 세포(Angelica gigas H4)를 현탁배양하여 면역 증강성 다당을 생산하기 위한 최적배지 설계 및 생산조건 최적화 연구를 수행하였다. 우선 세포의 성장에 가장 좋은 배지로서 Schenk and Hildebrandt(SH) 배지를 선택하여 배양한 결과 $25^{\circ}C$, 암배양조건에서 각각 15.8g DCW/l와 0.85 g/l의 세포 및 다당을 얻을 수 있었다. 한편 다당 생산용 배지의 개발을 위하여 여러 가지 식물세포배양용 배지 중에서 다당 생산능이 가장 높은 배지로서 Gamborg B5 배지를 선정하였으며 B5 배지를 변형하여 면역증강성 다당생산을 위한 최적배지(PPM)를 개발하였다. 그리고 최적배지를 이용한 당귀 세포증식과 다당 생산을 위한 최적 pH는 6.0-6.5, 배양온도는 $20^{\circ}C$, 그리 고 광조건보다는 암조건이 좋은 것으로 밝혀졌다. 상기 최적조건에서의 최대 세포증식과 다당생산은 각각 14.8 g/l, 1.5 g/l 이었다.

• 한국축산식품학회지
• /
• 제40권4호
• /
• pp.659-667
• /
• 2020

Muscle stem cells isolated from domestic animals, including cows and pigs, were recently spotlighted as candidates for the production of alternative protein resources, so-called cultured meat or lab-grown meat. In the present study, we aimed to optimize the in vitro culture conditions for the long-term expansion of pig muscle stem cells via the screening of various signaling molecules. Pig muscle stem cells were collected from the biceps femoris muscles of 3-d-old crossbred pigs (Landrace×Yorkshire×Duroc, LYD) and cultured in minimum essential medium-based growth media. However, the pig muscle stem cells gradually lost their proliferation ability and featured morphologies during the long-term culture over two weeks. To find suitable in vitro culture conditions for an extended period, skeletal muscle growth medium-2, including epidermal growth factor (EGF), dexamethasone, and a p38 inhibitor (SB203580), was used to support the stemness of the pig muscle stem cells. Interestingly, pig muscle stem cells were stably maintained in a long-term culture without loss of the expression of myogenic marker genes as determined by PCR analysis. Immunostaining analysis showed that the stem cells were capable of myogenic differentiation after multiple passaging. Therefore, we found that basal culture conditions containing EGF, dexamethasone, and a p38 inhibitor were suitable for maintaining pig muscle stem cells during expanded culture in vitro. This culture method may be applied for the production of cultured meat and further basic research on muscle development in the pig.

• Journal of Microbiology and Biotechnology
• /
• 제1권2호
• /
• pp.116-120
• /
• 1991

The transformant Bacillus subtilis ED213 carrying the pSO100 which cloned the cdd gene encoding cytidine deaminase (cytidine /2'-deoxycytidine aminohydrolase, EC 3.5.4.5, CDase) originated from wild type B. subtilis was cultivated in Spizizen minimal medium (SMM). To overcome poor expression of the cdd gene in SMM medium, the medium compositions and growth conditions were optimized. The optimized medium compositions and growth conditions were cytidine concentration of 80 mg/l, glycerol of 25 g/l, and $(NH_4)_2SO_4$ of 10 g/l, along with $37^{\circ}C$ and pH 7.0. The intracellular CDase production was increased 3 times from 1,000 unit/ml to 3,200 unit/ml, and extracellular CDase also increased from nearly undetectable amounts to 1,500 unit/ml. The cytidine concentration was signified as the most critical compositional factor for overproduction of CDase by increasing the cell density mainly in culture broth. The plasmids were more stable in cells that were grown in original SMM medium with stability of 90% compared to those grown in optimized SMM medium with stability of 80% after 48 hours cultivation. The most active amplification of plasmid was occurred in the logarithmic phase, which showed a value around four times higher than the initial copy number. In the exponential phase, the CDase production was closely related to the plasmid copy number along with the cell density. However, it was not accorded with cell density at the stationary phase.

• Biotechnology and Bioprocess Engineering:BBE
• /
• 제9권6호
• /
• pp.459-464
• /
• 2004

The objective of this study is to improve cephalosporin C (CPC) production byoptimization of medium and culture conditions. A statistical method was introduced to optimize the main culture medium. The main medium for CPC production was optimized using a statistical method. Glucose and corn steep liquor (CSL) were found to be the most effective factors for CPC production. Glucose and CSL were optimized to 2.84 and $6.68\%$, respectively. CPC produc­tion was improved $50\%$ by feeding of $5\%$ rice oil at day 3rd and 5th day during the shake flask culture of C acremonium M25. The effect of agitation speeds on CPC production in a 2.5-L bio­reactor was also investigated with fed-batch mode. The maximum cell mass (54.5 g/L) was obtained at 600 rpm. However, the maximum CPC production (0.98 g/L) was obtained at 500 rpm. At this condition, the maximum CPC production was improved about $132\%$ compared to the re­sult with batch flask culture.

• 생명과학회지
• /
• 제9권1호
• /
• pp.22-28
• /
• 1999

A bacterium producing exo-inulinase was isolated from soil and identified Pseudomonas sp. and named as Pseudomonas sp. NO5. The optimal culture conditions for the efficient production of exo-inulinase from Pseudomonas sp. NO5 were obtained by cultivating with the medium 1$\%$ sucrose, 0.5$\%$ yeast extract, 0.5$\%$ $(NH_4)_2$$HPO_4$, 0.05$\%$ $MgSO_4$ㆍ$7H_2$0, 0.001$\%$ and $FeSO_4$ㆍ$7H_2$0 at $37^{\circ}C$ in initial pH 7.0 for 20 hours. The enzyme was induced maximally in the presence of sucrose or inulin at early stationary phase about 20 hour after cultivation.

• 한국염색가공학회지
• /
• 제26권3호
• /
• pp.237-241
• /
• 2014

The aim of this study is to develop an eco-friendly reduction process of indigo as an alternative choice. Hansenula misumaiensis was used and their reducing activity toward synthetic indigo as well as natural indigo was estimated by dyeing test in terms of indigo dye uptake. The changes in K/S value and pH were monitored on the time-based measurements. Also, reduction duration was evaluated. On the basis of the results described in this study, it was confirmed that Hansenula misumaiensis reduced indigo. Reducing power of Hansenula misumaiensis reached to maximum in two days. It can be possible to develop eco-friendly process of indigo reduction using Hansenula misumaiensis by the optimization of strain culture conditions and the optimization of reduction conditions.

• 한국미생물·생명공학회지
• /
• 제29권2호
• /
• pp.110-114
• /
• 2001

가축 사료첨가용 probiotics용 균주를 얻기 위해 특히 내열성이 높아 가공사료에 적합한 Bacillus sp.를 위주로 하여 가축에게 유용한 효소인 phytase를 비롯하여 protease, cellulase, xylanase, amylase의 활성을 모두 나타내는 4-3 균주를 얻어 동정한 결과 Bacillus subtilis로 밝혀졌으며, 이를 B. subtilis 4-3으로 명명하였다. 본 균주를 원료사료에 순수배양하여 사료의 항영양인자인 phytic acid 분해율을 검토한 결과 대두박 및 쌀겨에 있어서는 phytic acid 분해율이 낮았으나, 밀기울의 경우 80.63%로 상대적으로 높은 phytic acid 분해율을 나타내었다. 원료 사료를 이용한 균주의 생산 조건은 대두박 1%(w/v)와 당밀 2%(w/v)가 가장 적합한 균주 생산을 위한 배지조성으로 검토되었다.

• 한국동물생명공학회지
• /
• 제38권2호
• /
• pp.70-76
• /
• 2023

Background: Despite considerable technological advancements, polyspermy remains a significant challenge in in vitro fertilization (IVF) procedures in pigs, disrupting normal embryonic development. Here, we aimed to determine whether optimal fertilization conditions reduce the polyspermy incidence in pigs. Methods: In vitro-matured oocytes were co-incubated with sperm according to a modified two-step culture system. Results: In the first experiment, oocytes were briefly co-incubated with sperm, washed in IVF medium, and then moved to fresh IVF medium for 5 or 6 h. Although the 6 h sperm-free cultured group had a higher penetration rate than the 5 h cultured group, the polyspermy rate significantly increased in the 6 h sperm-free cultured group. The gamete co-incubation period was either 20 or 40 min. The 40 min cultured group had a higher rate of blastocyst formation and number of total cells in blastocysts than the 20 min cultured group. In experiment 2, oocytes were inseminated with sperm separated by Pecroll treatment. Percoll treatment increased the rate of oocyte penetration and blastocyst formation compared to the control. In experiment 3, fertilized oocytes were cultured in 25 µL microdroplets (10 gametes/drop) or 500 µL (100 gametes/well) of culture medium in 4-well plates. The large volume of medium significantly reduced the number of dead oocytes and increased the rate of blastocyst formation compared to the small volume. Conclusions: Collectively, these results demonstrate that various fertilization conditions, including modified co-culture period, active sperm separation, and culture medium volume, enhance fertilization efficiency and subsequent embryonic development by decreasing polyspermy occurrence.

• 한국식생활문화학회지
• /
• 제23권2호
• /
• pp.243-251
• /
• 2008

This purpose of this study was to develop a functional muffin by adding yam powder in the shape of a muffin as a partial surrogate for wheat flour. The yam has been found to be effective for liver and kidney function, as well as the digestion of protein, since it produces glucuronic acid in the body. Therefore, the purpose of this study was to determine the optimal mixing conditions of yam muffins by adjusting the amounts yam powder, butter, and sugar. The mixing conditions for the yam muffins included 3 categories: yam powder $(X_1)$, sugar $(X_2)$, and butter $(X_3)$ by Central Composite Design (CCD) which was optimized by Response Surface Methodology (RSM). The effects of the three variable additions on muffin quality were examined via physical and chemical experiments, such as the analysis of texture (hardness, cohesiveness, springiness, gumminess), coloration (lightness, redness, yellowness), and height. Lastly, we performed a sensory test, which revealed significant findings for gumminess, color, appearance, flavor, softness (p<0.05), redness, and overall quality (p<0.01). Consequently, the optimal mixing rate which best satisfied the sensory items were 34.35g of yam powder, 80.15 g of sugar, and 80.55 g of butter.

• 미생물학회지
• /
• 제41권2호
• /
• pp.130-134
• /
• 2005

Monascus purpureus KCCM 60016을 이용하여 다량의 색소생산 변이주를 개발하고 생산조건에 대하여 조사하였다. 자외선 조사로 M. purpureus KCCM 60016으로부터 돌연변이를 유도한 결과, 색소생성이 우수한 P-57 변이주를 분리하였다. 분리된 P-57 변이주는 고체 배양에서 현미를 기질로 했을 때 색소생성이 가장 우수한 것으로 나타났으며, 색소생성의 최적 배양조건은 배양온도 $30^{\circ}C,\;90\%$의 배양습도에서 30일간 배양이 가장 우수했다. 이상의 조건에서 배양한 적색색소, 오렌지색소와 황색색소의 양은 각각 160.0 unit, 193.6 unit와 141.6 units-나타났다.