• Preventive Nutrition and Food Science
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• 제18권4호
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• pp.273-279
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• 2013

Protease widely exists in the digestive tract of animals and humans, playing a very important role in protein digestion and absorption. In this study, a high protease-producing strain Planomicrobium sp. L-2 was isolated and identified from the digestive tract of Octopus variabilis. The strain was identified by physiological and biochemical experiments and 16S rDNA sequences analysis. A protease was obtained from the strain Planomicrobium sp. L-2 through ammonium sulfate precipitation, dialysis and enrichment, DEAE-Sephadex A50 anion-exchange chromatography, and Sephadex G-100 gel chromatography. The molecular weight and properties of the protease were characterized, including optimum temperature and pH, thermal stability, protease inhibitions and metal ions. According to our results, the protease from Planomicrobium sp. L-2 strain designated as F1-1 was obtained by three-step separation and purification from crude enzyme. The molecular weight of the protease was 61.4 kDa and its optimum temperature was $40^{\circ}C$. The protease F1-1 showed a broad pH profile for casein hydrolysis between 5.0~11.0. No residual activity was observed after incubation for 40 min at $60^{\circ}C$ and 60 min at $50^{\circ}C$. F1-1 protease was inhibited by $Mn^{2+}$, $Hg^{2+}$, $Pb^{2+}$, $Zn^{2+}$, and $Cu^{2+}$ ions, as well as PMSF, indicating that the protease F1-1 was a serine protease. Additionally, research basis provided by this study could be considered for industrial application of octopus intestinal proteases.

• 한국미생물·생명공학회지
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• 제30권1호
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• pp.26-32
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• 2002

저염상태에서 속성 발효를 통한 멸치 어간장의 대량 생산을 위하여 시중 어간장에서 단백질 분해효소 생산이 우수한 균주를 분리 동정하여, B. amyloliquefaciens HTP-8로 명명하였다. 효소생산을 위한 배양 최적온도, 초기 pH, 그리고 배양시간은 각각 $30^{\circ}C$, pH 7.0과 3일이었다. Jar fermenter 배양시 pH를 7.0으로. 조절한 경우가 조절하지 않은 경우에 비해 효소활성이 최대에 이르는 시간이 단축되었다. 효소의 부분정제는 조효소액을 80% ammonium sulfate에 의한 염석과 CM-Sephadex C-50을 이용하여 정제한 결과, 수율이 0.4%, 정제배수가 43.0배였다. 정제효소의 최적 pH와 온도는 pH 10.0과 $50^{\circ}C$였으며, pH 7.0~l2.0의 pH 범위와 $40^{\circ}$ 이하에서 안정하였다. 금속이온 중 $Ag^{＋}$ /, $Ba^{2＋}$ /에 의하여 효소활성이 저해되었다. 한편, 본 효소는 PMSF에 의하여 선택적으로 활성이 억제됨으로써 활성부위에 serine을 가진 serine protease에 속하는 것으로 판단되었다.

• Proceedings of the National Institute of Ecology of the Republic of Korea
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• 제2권4호
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• pp.229-234
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• 2021

Iris dichotoma Pall. found on Daechung Island in Korea has been designated as an endangered species. To aid in conservation efforts of this species, this study investigated its germination characteristics and seed dormancy type. Four sets of seeds were incubated at different temperatures (4/1℃, 15/6℃, 20/10℃, and 25/15℃). One set of seeds was cold stratified (4 weeks at 4/1℃). The final germination rate and mean germination time showed that the optimal germination temperature was 25/15℃. Final germination rates were ~70%, showing no significant difference among temperature treatments. However, mean germination time were significantly different among all temperature treatments except for 4/1℃. Mean germination time for seeds with temperature treatments of 15/6℃, 20/10℃, and 25/15℃ were 3.2, 2.1, and 1.5 weeks, respectively. At 25/15℃, the mean germination time was half of that at 15/6℃. Seeds of I. dichotoma had fully developed embryos at the time of dispersal. No additional growth of the embryo was observed. Cold stratification did not affect the final germination rate or the mean germination time. This study shows that seeds of I. dichotoma have no physiological or morphological dormancy, unlike other members of the Iris genus known to have seed dormancy that needs a relatively high incubation temperature (≥25/15℃) for mass propagation to occur. These results will be useful for understanding ecophysiological mechanisms related to the species' habitat. They are also useful for mass propagation of I. dichotoma for the purpose of conserving this endangered species.

• 미생물학회지
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• 제52권4호
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• pp.398-405
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• 2016

이전의 연구에서, bacterioferritin comigratory protein (BCP)을 인코딩하는 Schizosaccharomyces pombe의 구조유전자를 shuttle vector인 pRS316에 클로닝하여 BCP 과잉발현 플라즈미드인 pBCP10을 제조한 바 있다. 본 연구에서는, 플라즈미드 pBCP10을 사용하여 고온 스트레스에 대한 BCP의 열저항적 성질을 평가하였다. 대수기의 초기까지 성장시킨 S. pombe 세포의 배양 온도를 $30^{\circ}C$에서 $37^{\circ}C$나 $42^{\circ}C$로 전이시키는 경우, pBCP10 함유 S. pombe 세포가 벡터 대조 세포보다 $37^{\circ}C$와 $42^{\circ}C$ 모두에서 유의하기 더 잘 성장하였다. 높은 배양 온도로 전이한 뒤 6시간에서, pBCP10 함유 S. pombe 세포가 벡터 대조 세포보다 낮은 활성산소종(ROS)과 일산화질소(NO)의 지표로 측정된 아질산염(nitrite) 함량을 갖고 있음이 확인되었다. 온도 전이 뒤에, 총 글루타치온(total glutathione) 함량과 총 수퍼옥사이드 디스뮤타제(superoxide dismutase) 활성은 대응되는 벡터 대조 세포보다 pBCP10 함유 S. pombe 세포에서 현저하게 높다는 사실도 확인되었다. 종합하면, S. pombe BCP는 열저항적 역할을 보유하는 데, 활성산소종과 일산화질소에 대한 하강시키는 활성과 총 글루타치온과 수퍼옥사이드 디스뮤타제 등의 항산화 성분들을 상승시키는 활성, 즉 종합적으로 열안정성을 유지하는 활성에 근거하는 것으로 추정되었다

• 한국환경보건학회지
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• 제27권1호
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• pp.75-82
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• 2001

Microorganisms can attach firmly to food contact surface material and the resitance of adherent bacteria differ markedly from planktonic cells. Therefore, adherent cells are a potential contamination problem to the food preparation because of their high resistance. to sanitation and heat treatment. This study was carried out in order to investigate growth and decontamination of Listeria monocytogenes attached to stainless steel, glass and plastic. Listeria monocytogenes cells could attach to all types of surface at three temperatures after contact times for 24 hrs. The numbers of adherent cells were greater at higher temperatures, but not increased with incubation time. When recovery of adherent cells was investigated, after 24 grs, the numbers of adherent cells were about 10$^{7}$ , 10$^{10}$ , 11$^{11}$ at 4$^{\circ}C$, $25^{\circ}C$, 3$0^{\circ}C$ repectively. Planctonic cells decreased by 2 log cycles after exposure to the domestic sanitizer. Adherent cells showed high resistance to domestic sanitizers and that was dependent upon surface materials studied, being greatest on plastic followed by stainless steel and glass. Adherent cells were more resistant to heat treatment than planktonic cells. When adherent cells were exposed to the temperature of 5$0^{\circ}C$, 55$^{\circ}C$, 57.5$^{\circ}C$ for 10 min, their populations did not decrease significantly. When the temprature increased to 6$0^{\circ}C$, cells attached to all types of surfaces were completely inactivated for 10 min.

• Asian-Australasian Journal of Animal Sciences
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• 제26권11호
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• pp.1651-1658
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• 2013

Nitrate contamination in ground and surface water is an increasingly serious environmental problem and only a few bacterial strains have been identified that have the ability to remove nitrogen pollutants from wastewater under thermophilic conditions. We therefore isolated thermophilic facultative bacterial strains from wood chips that had been composted with swine manure under aerated high temperature conditions so as to identify strains with denitrifying ability. Nine different colonies were screened and 3 long rod-shaped bacterial strains designated as SG-01, SG-02, and SG-03 were selected. The strain SG-01 could be differentiated from SG-02 and SG-03 on the basis of the method that it used for sugar utilization. The 16S rRNA genes of this strain also had high sequence similarity with Geobacillus thermodenitrificans $465^T$ (99.6%). The optimal growth temperatures ($55^{\circ}C$), pH values (pH 7.0), and NaCl concentrations (1%) required for the growth of strain SG-01 were established. This strain reduced 1.18 mM nitrate and 1.45 mM nitrite in LB broth after 48 h of incubation. These results suggest that the G. thermodenitrificans SG-01 strain may be useful in the removal of nitrates and nitrites from wastewater generated as a result of livestock farming.

• The Journal of Advanced Prosthodontics
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• 제9권6호
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• pp.453-462
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• 2017

PURPOSE. The purpose of this study was to investigate the in vitro cytotoxicity of thermoplastic denture base resins and to identify the possible adverse effects of these resins on oral keratinocytes in response to hot water/ food intake. MATERIALS AND METHODS. Six dental thermoplastic resin materials were evaluated: three polyamide materials (Smile tone, ST; Valplast, VP; and Luciton FRS, LF), two acrylic materials (Acrytone, AT; and Acryshot, AS), and one polypropylene resin material (Unigum, UG). One heat-polymerized acrylic resin (Vertex RS, RS) was chosen for comparison. After obtaining extracts from specimens of the denture resin materials (${\phi}=10$ mm and d=2 mm) under different extraction conditions ($37^{\circ}C$ for 24 hours, $70^{\circ}C$ for 24 hours, and $121^{\circ}C$ for 1 hour), the extracts (50%) or serial dilutions (25%, 12.5%, and 6.25%) in distilled water were co-cultured for 24 hours with immortalized human oral keratinocytes (IHOKs) or mouse fibroblasts (L929s) for the cytotoxicity assay described in ISO 10993. RESULTS. Greater than 70% viability was detected under all test conditions. Significantly lower IHOK and L929 viability was detected in the 50% extract from the VP ($70^{\circ}C$) and AT ($121^{\circ}C$) samples (P<.05), but only L929 showed reduced viability in the 50% and 25% extract from LF ($37^{\circ}C$) (P<.05). CONCLUSION. Extracts obtained from six materials under different extraction conditions ($37^{\circ}C$, $70^{\circ}C$, and $121^{\circ}C$) did not exhibit severe cytotoxicity (less than 70% viability), although their potential risk to oral mucosa at high temperatures should not be ignored.

• Journal of the Korean Wood Science and Technology
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• 제34권1호
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• pp.68-78
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• 2006

돈분뇨 분해 장치의 발효 속도를 가속화시키는 토착미생물의 확보를 위하여 우리나라 각 지역에서 수집한 균주 중 우수한 균주를 선별하고 168 rDNA 증폭에 의한 동정을 실시하여 돈분뇨 분해에 관여하는 균주의 특성을 확인하기 위해 본 연구에서는 총 23장소(서울, 충청, 강원, 전북, 전념, 경남 지역)에서 28종류의 부숙 퇴비를 수집하여 생균수를 조사한 결과 대체적으로 $1{\times}10^5{\sim}10^8$의 분포 밀도를 보였다. 분리한 균주의 효소 생산 역가결과는 $30^{\circ}C$에서 보다 $55^{\circ}C$에서 배양된 고온호기성박테리아가 상대적으로 높은 섬유소분해효소와 전분분해효소 생산력을 보였고, genomic DNA의 추출에 의한 168 primer로 증폭하여 염기서열을 결정한 결과 돈분뇨 발효에 관여하는 고온호기성박테리아는 15종으로 동정되었다. 그 중 미생물제재를 제조하기 위하여 효소 역가 면에서도 우수성을 보이고 내생 포자를 형성하는 균주는 Bacillus subtihs로 확인되었다.

• KSBB Journal
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• 제25권5호
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• pp.453-458
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• 2010

본 연구에서는 효소의 재활용성과 안정성을 확보하기 위해 자석으로 분리가 가능한 polyaniline nanofiber를 개발하였다. 개발된 고정화 효소는 상온에서 8일 동안 90% 이상의 활성도를 보유하였으며 온도가 높은 $55^{\circ}C$에서는 60% 이상의 활성도를 보유하여 안정성의 유지현상을 보였다. 개발된 고정화 효소는 자석으로 분리가 가능하였으며 이 효소를 이용하여 curdlan, agarose, cellulose, 및 미역을 분해한 결과 포도당을 생산하였으며 curdlan을 분해시킨 경우에는 분해 속도가 1.2 g/L/h로 나타나 다른 다당류에 비해 3-10배 이상 빠른 속도를 나타내었다. 고정화 효소를 반복하여 사용하는 경우 10번 반복 사용했을 때 75% 이상의 활성도를 유지하는 것으로 측정되었다. 젖은 미역 줄기를 10 g/L를 분해하기 위하여 5 mg의 고정화 효소를 사용한 결과 24시간 만에 1 g/L의 glucose를 생산하였다.

• 한국축산식품학회지
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• 제42권6호
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• pp.942-952
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• 2022

To establish a pre-plating method of chicken satellite cells with high purity, pre-plating was performed under culture conditions of 37℃ and 41℃, and the pre-plating time was set from a total of 3 hours to 6 hours in consideration of the cell attachment time. The purity of the cells was confirmed by staining paired box protein 7 (Pax7) after proliferation, and Pax7 expression was the highest in culture flasks shaken for 2 hours after incubation at 41℃ for 2 hours to prevent the attachment of satellite cells (p<0.05). Also, when pre-plating and proliferation were performed at 37℃ and 41℃, the Pax7 expression rate was higher at 41℃. The differentiation capabilities of the three groups (T3, T6, and T7) with high Pax7 expression were compared and the fusion index (%) and myotube formation area (%) determined by myosin heavy chain (MHC) staining was calculated. The T6 and T7 groups, which were cultured at 41℃, showed significantly higher values than the T3 group (p<0.05). There was no significant difference in the expression of Pax7 and MHC between the T6 and T7 groups (p>0.05). These results suggest that pre-plating at 41℃ for a total of 4 hours was the most efficient in terms of cost and time for purifying chicken satellite cells for cultured meat.