고온내성 연료용 알코올 효모균주 Saccharomyces cerevisiae KNU5377에서 HSF1 유전자의 변이주 구축

김일섭,윤혜선,최혜진,손호용,유춘발,김종국,진익렬,Kim Il-Sup,Yun Hae-Sun,Choi Hye-Jin,Sohn Ho-Yong,Yu Choon-Bal,Kim Jong-Guk,Jin Ing-Nyol Korean Society of Life Science 2006 생명과학회지 Vol.16 No.3

출아효모인 Sacharomyces cerevisiae S288C균주를 이용한 효모의 게놈이 완성된 후 S. cerevisiae는 다양한 연구 모델로 이용되어져 왔다. 현재까지 효모를 이용한 기능 유전체학 측면에서의 연구는 laboratory strainin인 S288C 균주 또는 그 유래의 균주들이다. 그러나 자연에서 분리된 효모 또는 산업적으로 이용되어지고 있는 S. cerevisiae의 유전학 측면에서의 연구는 낮은 포자형성률 및 형질전환률, 그리고 S288C 균주와의 게놈상의 상이성 때문에 거의 이루어지지 않고 있다. 여기서 우리 연구진은 자연에서 분리된 Saccharomyces cerevisiae KNU5377 균주를 이용하여 random spore analysis를 통해 MATa 및 $MAT{\alpha}$ 타입의 각각의 haploid cell을 분리 후 이미 보고된 KanMX module를 가지고 round PCR기법에 의한 short flanking homology 기법을 이용하여 전사조절인자인 HSF1 유전자가 치환된 변이주를 구축할 수 있었다. 덧붙여, 모든 유전자에 이 기법을 적용할 수는 없다는 것을 확인하였다. 앞으로 이 변이주를 통해 기능 유전체학적인 측면에서 이 유전자의 스트레스와의 관련성을 연구하고자 한다. HSF1 is the heat shock transcription factor in Saccharomyces cerevisiae. S. cerevisiae KNU5377 can ferment at high temperature such as $40^{\b{o}}C$. We have been the subjects of intense study because Hsf1p mediates gene expression not only to heat shock, but to a variety of cellular and environmental stress challenges. Basing these facts, we firstly tried to construct the hsf1 gene-deleted mutant. PCR-method for fast production of gene disruption cassette was introduced in a thermotolerant yeast S. cerevisiae KNU5377, which allowed the addition of short flanking homology region as short as 45 bp suffice to mediate homologous recombination to kanMX module. Such a cassette is composed of linking genomic DNA of target gene to the selectable marker kanMX4 that confers geneticin (G418) resistance in yeast. That module is extensively used for PCR-based gene replacement of target gene in the laboratory strains. We describe here the generation of hsf1 gene disruption construction using PCR product of selectable marker with primers that provide homology to the hsf1 gene following separation of haploid strain in wild type yeast S. cerevisiae KNU5377. Yeast deletion overview containing replace cassette module, deletion mutant construction and strain confirmation in this study used Saccharomyces Genome Deletion Project (http:://www-sequence.standard.edu/group/yeast_deletion_project). This mutant by genetic manipulation of wild type yeast KNU5377 strain will provide a good system for analyzing the research of the molecular biology underlying their physiology and metabolic process under fermentation and improvement of their fermentative properties.

심한 부종을 동반한 분비성 뇌수막종 - 증례보고 -

김일섭,이형진,이진석,양지호,이일우,강준기,Kim, Il-Sup,Lee, Hyung-Jin,Lee, Jin-Suck,Yang, Ji-Ho,Lee, Il-Woo,Kang, Joon-Ki 대한신경외과학회 2001 Journal of Korean neurosurgical society Vol.30 No.1

The authors report a 42-year-old woman presented with a falx meningioma with disproportionately severe peritumoral edema. Histological examination including immunohistochemical staining resulted in the diagnosis of secretory meningioma. In addition to tumor size, edema could not be explained by location, growth rate, vascular involvement, or other factors. We conclude that secretory meningiomas may possess an innate ability to cause brain edema.

Increased Antioxidative Activities against Oxidative Stress in Saccharomyces cerevisiae KNU5377

Il-Sup Kim(김일섭),Hae-Sun Yun(윤혜선),Ji Young Yang(양지영),Oh-Seok Lee(이오석),Heui-Dong Park(박희동),Ingnyol Jin(진익렬),Ho-Sung Yoon(윤호성) 한국생명과학회 2009 생명과학회지 Vol.19 No.4

산화적 스트레스는 정상적인 대사 과정뿐만 아니라 외부적인 환경에 노출 되었을 때 일어나는 것으로 잘 알려져 있다. 이러한 스트레스를 극복하기 위해 생물체들은 각자의 시스템에 맞게 다양한 항산화 시스템을 진화 발전시켜 왔다. Saccharomyces cerevisiae KNU5377 균주는 고온뿐만 아니라 다양한 스트레스에 대해 내성을 가짐을 확인하였다. 대부분의 스트레스는 궁극적으로는 산화적 스트레스로 귀결된다. 이러한 측면에서 본 연구는 KNU5377 균주가 어떠한 시스템에 의해서 다른 균주보다 스트레스 내성을 가지는지를 밝히기 위해 접근하였다. 수행된 연구결과에서 KNU5377 균주는 항산화 시스템과 밀접하게 관련된 단백질(superoxide dismutase, thioredoxin system, heat shock proteins)과 항산화 관련 물질(trehalose)을 과발현함을 확인하였다. 그러나 이러한 단백질들이 어떠한 조절 시스템에 의해서 균주 특이적인 발현 양상을 보이는지는 현재까지 확인되지 않고 있다. 본 연구는 KNU5377 균주 그 자체의 중요성과 함께 균주 내의 스트레스 내성과 관련된 유용한 유전자를 탐색하여 더욱 우수한 유전자원을 발굴하는데 기여 할 것으로 보인다. Oxidative stress is a consequence of an imbalance of the defense system against cellular damage generated by reactive oxygen species (ROSs) such as superoxide anions (menadione; MD). Most organisms have evolved a variety of defense systems to protect cells from adverse conditions. In order to evaluate stress tolerance against oxidative stress generating MD, comparative analyses of antioxidant capacity, or free radical scavenger ability, were performed between S. cerevisiae KNU5377 (KNU5377) and three wild-type S. cerevisiae strains. In a medium containing 0.4 mM MD, the KNU5377 strain showed higher cell viability and antioxidant ability, and contained higher levels of trehalose, superoxide dismutase, thioredoxin system, glucose-6-phosphate dehydrogenase, and some heat shock proteins. The KNU5377 strain also produced a lower level of oxidative stress biomarker than the other three yeast strains. These results indicate that S. cerevisiae KNU5377 has a higher level of tolerance to oxidative stress due to the increased expression of cell rescue proteins and molecules, thus alleviating cellular damage more efficiently than other S. cerevisiae strains.

환경 스트레스에 의한 세포 내 신호의 이동 경로와 유전적 조절

김일섭 ( Il Sup Kim ),김현영 ( Hyun Young Kim ),강홍규 ( Hong Gyu Kang ),윤호성 ( Ho Sung Yoon ) 한국환경생물학회 2008 환경생물 : 환경생물학회지 Vol.26 No.4

송과체 부위 종양 수술적 해부 및 접근방법

김일섭 ( Il Sup Kim ),이상원 ( Sang Won Lee ) 대한뇌종양학회·대한신경종양학회·대한소아뇌종양학회 2006 대한뇌종양학회지 Vol.5 No.1

The pineal region tumor is one of the difficult ones to exposure and remove. Selecting an operative approach to the pineal region tumor involving the third ventricle requires an understanding of the microsurgical anatomy of the posterior third ventricle. There are variable surgical approaches to the pineal region. The approaches suitable for reaching the pineal region are the infratentorial supracerebellar, occipital transtentorial, posterior transcallosal, posterior transcortical and combined supra/infratentorial-transsinus approach. Considerations important in selecting one of these surgical approaches are reviewed from the anatomical viewpoint.

부비동염 수술 후 시행한 뇌하수체 샘종 제거를 위한 경접형동 접근법의 임상 결과

김동섭(Dong Sup Kim),박찬순(Chan-Soon Park),김영일(Young Il Kim),김일섭(Il Sup Kim),성재훈(Jae Hoon Sung),양승호(Seung Ho Yang) 대한두개저학회 2019 대한두개저학회지 Vol.14 No.1

Background: Transsphenoidal surgery (TSS) is the mainstay of treatment for sellar and parasellar lesions. Nasal and paranasal sinus inflammation is considered a contraindication to TSS because of possible transcranial spread of infection. The objective of this study was to determine outcomes of TSS after operations for rhinosinusitis. Methods: Medical records and radiological findings of 8 consecutive patients who underwent TSS for pituitary adenomas following operations for rhinosinusitis were reviewed. Results: These patients consisted of 2 men and 6 women, with a mean age of 56.8 years (range, 34-74 years). Their presenting symptoms were visual defect (4 patients) and headache (4 patients) from neurosurgeon’s aspect. Rhinology examination revealed a preoperative Lund-Mackay score of 1 to 6 with involvement of sphenoid and ethmoid sinuses in 8 and 4 patients, respectively. Fungal infection was confirmed in 3 patients intraoperatively. The median interval between rhinosinusitis surgery and TSS was 55.8 days (range, 7-155 days). The median duration of antibiotics administration was 32.9 days (range, 9-84 days) after the rhinosinusitis surgery. Intraoperative tearing of suprasellar cistern and cerebrospinal fluid (CSF) leakage occurred in 5 patients during TSS for pituitary adenomas. Postoperative meningitis was noted in 2 patients and treated with further antibiotics administration. Conclusions: It is important to treat rhinosinusitis issues preoperatively for TSS. Intraoperative CSF leakage could increase the risk of postoperative meningitis in patients with rhinosinusitis even though it has been treated surgically and medically. Collaboration between rhinologist and neurosurgeon is mandatory for patients’ safety.

40℃ 48시간 에탄올발효 과정 중 일어나는 Saccharomyces cerevisiae KNU5377의 생리변화

Sun-Hye Kwak(곽선혜),Il-Sup Kim(김일섭),Kyung-Hee Kang(강경희),Jung-Sook Lee(이정숙),Ingnyol Jin(진익렬) 한국생명과학회 2011 생명과학회지 Vol.21 No.1

포도당 20%와 효모엑기스가 함유된 배지에서 40oC에서 48시간에 걸친 에탄올발효 과정 중에 일어나는 고온성 알코올 발효 효모균주 Saccharomyces cerevisiae KNU5377의 세포 내에서 일어나는 생리적 변화를 살펴보고자 했다. 그 결과, 이 발효 효모균주는 40℃ 48시간의 발효로써 11.4% alcohol을 생성하여, 고온발효인데도 불구하고 우수한 발효능을 가진 것을 확인할 수 있었다. 또한 12시간 발효과정동안 배지의 pH가 시작단계의 pH 6.0이 4.1로 내려갔으며, 이후 에탄올발효가 완성될 때까지 거의 변화하지 않고 이 값으로 유지되었다. 발효 12시간이 지나면서 세포막의 지방산의 조성은 불포화지방산인 C16:1 (palmitoleic acid)가 포화지방산인 C16:0 (palmitic acid)보다 1.5배나 증가하였으며, C18:1보다는 2배 가까이 증가하여 구성되어 있었다. 48시간 배양한 세포의 2차 전기영동(2-D)을 통한 세포내 단백질의 발현정도를 proteomics분석법으로 살펴본 바, phosphoglycerate kinase가 가장 크게 발현했음을 Mass Spectrometry를 통해 알 수 있었다. 그 뒤를 이어 adenylate kinase, Cys3p, Tdh3p, translational elongation factor 등이 크게 발현된 것을 알게 되었다. 이들은 직간접적으로 해당과정에 관여하는 인자들 이어서, 고온 장시간에 걸친 에탄올발효를 하는 이 발효 효모균주의 세포에게는 생존과 에탄올발효를 위하여 해당과정 관여 인자가 중요한 역할을 하고 있다는 것을 강력히 시사하였다. In this study, physiological changes in a thermotolerant yeast Saccharomyces cerevisiae KNU5377 cell exposed to 48-hour alcohol fermentation at 40℃ were investigated. After 12 hours of alcohol fermentation at 40℃, the C16:1 unsaturated acid of plasma membrane increased to 1.5 times more than the C16:0 saturated fatty acid, and to about 2 times more for the C18:1 unsaturated fatty acid. Fermentation at both 30℃- and 37℃- fermentation showed the same pattern as that done at 40℃. The pH of the alcohol-fermentation medium was reduced to pH 4.1 from a starting pH of 6.0 through the 12-hr fermentation and then maintained this level during the continuing fermentation. With the process of fermentation, the remaining glucose was reduced, but its amount remaining during the 40℃-fermentation was less reduced than those fermented at 30℃ and 37oC. In the study investigating the changing pattern of cellular proteins in the alcohol-fermenting cells, the SDS-PAGE and 2-D data indicated the most expressed dot was phosphoglycerate kinase, which is one enzyme involved in glycolysis. Why this enzyme was most expressed in the cells exposed to unfavorable conditions such as high temperature, increasing concentration of produced alcohol and long time exposure to other stress factors remains unsolved.

큰 뇌하수체 선종의 경접형동 접근법의 치료 경험

양승호(Seung-Ho Yang),김일섭(Il Sup Kim),홍재택(Jae Taek Hong),성재훈(Jae Hoon Sung),손병철(Byung Chul Son),이상원(Sang Won Lee) 대한두개저학회 2007 대한두개저학회지 Vol.2 No.1

Objective The purpose of this study is to determine the feasibility of transsphenoidal approach (TSA) and staged or repeated TSA for large pituitary adenoma (LPA) and remnant or recurrent PA. Methods Eighteen LPAs underwent TSA surgery from February 2002 to December 2006. Eleven cases were non-functioning adenomas, 5 cases were growth hormone secreting adenomas and 2 cases were pituitary apoplexy. Mean size of tumor was 38x28x30mm. All patients were initially operated via TSA. Results Total removal was performed in 8 patients (44.4%). Subtotal removal was done in 8 patients (44.4%) by the initial TSA, two patients in which underwent repeated TSA due to postoperative tumor bleeding. The visual and hormonal symptoms were improved in 13 patients (72.2%) after the surgery. Six patients out of 8 with subtotal removal underwent the staged operation. Two patients with tumor recurrence underwent repeated TSA, 18 and 26 months respectively after the initial TSA. All patients with growth hormone-secreting PA achieved the normalization of growth hormone. Conclusion The staged and repeated TSA is one of the feasible option for the treatment of LPA and residual or recurrent PA with acceptable surgical morbidity and mortality.

임상 : 상추체사대부 수막종 수술 후 삶의 질 평가

김문석 ( Moon Seok Kim ),양승호 ( Seung Ho Yang ),김일섭 ( Il Sup Kim ),홍재택 ( Jae Taek Hong ),성재훈 ( Jae Hoon Sung ),손병철 ( Byung Chul Son ),이상원 ( Sang Won Lee ) 대한뇌종양학회 2012 대한뇌종양학회지 Vol.11 No.1

Objectives: This study was performed to assess the quality of life (QoL) of the patients with petroclival meningioma treat-ed with surgery with/without radiotherapy. Methods: From 2005 to 2010, 12 patients with newly diagnosed petroclival meningioma underwent surgery at a single in-stitution. The status of preoperative and postoperative (6 months) QoL was assessed using EORTC QLQ-C30 and Brain Cancer Module (BN20). Results: The most frequent preoperative symptoms were headache and dizziness. Total, subtotal, and partial resections were achieved in five, two, and five patients, respectively. Radiation therapy was performed in four patients with subtotal or par-tial resection. The postoperative complications were cranial nerve palsy (n=3), hemiparesis (n=2), and meningitis (n=2). Delayed hydrocephalus was found in two patients treated with additional radiosurgery. Motor dysfunction, bladder control, nausea/vomiting, dyspnea, insomnia, appetite loss, and constipation deteriorated and worsened the status of patients` QoL. However, headache, future uncertainty, and visual disorder improved. Conclusion: Postoperative QoL of the patients with petroclival meningiomas could be associated with surgery with/with-out radiation-related morbidity. Besides the treatment-associated morbidity and mortality rates, QoL issues should be eval-uated in the research of skull base tumors in the future.

폐양액을 이용한 관비재배시 양액의 농도가 파프리카의 생육과 과실 품질에 미치는 영향

장성호(Cheng Hao Zhang),임용섭(Yong Sup Lim),강호민(Ho-Min Kang),김일섭(Il Seop Kim) 한국원예학회 2010 원예과학기술지 Vol.28 No.1

파프리카 암면재배시 배출된 폐양액의 EC는 공급원액의 ±5%와 pH는 ±15% 수준으로 안정적이었고, 양이온 중 암모니아태 질소(NH₄-N)는 감소하였고, Mg는 증가하였으며, 음이온 중에는 SO₄-S가 증가하였다. 폐양액과 고추용 관비양액으로 재배한 파프리카의 초장과 엽수 엽두께는 차이가 없었으나, 경경과 엽면적 지수는 폐양액 2.0dS · m?¹처리에서 우수하였다. 생육기간 중 엽록소 함량은 1.5와 2.0dS · m?¹ 폐양액 처리가 관비양액 처리보다 높았으며, 총 수량과 과실수 그리고 과경은 2.0dS · m?¹폐양액 처리에서 우수하였다. 수확한 과실의 배꼽썩음과는 폐양액 처리에서는 모두 발생하지 않은 반면 관비용 양액처리에서는 5.3%가 발생하였다. 파프리카 잎의 무기물 함량은 큰 차이를 보이지 않았으나, 배꼽썩음과의 원인 원소인 Ca은 폐양액 처리구에서 K는 관비용 양액 처리구에서 높았으며, 엽록소 구성 원소인 Mg은 폐양액 2.0dS · m?¹에서 가장 높았다. 수확 한 파프리카 과실를 크기별로 분류했을때 100-150g 수준의 중과는 폐양액 2.0dS · m?¹과 관비용 양액 처리에서 52% 이상으로 높았으며 100g 이하의 소과는 관비용 양액 처리가, 150g이상의 대과는 폐양액 2.0dS · m?¹처리에서 가장 많았다. 따라서 폐양액 농도를 관비용 양액보다 25% 높은 농도로 공급할 경우 식물의 생육과 과실의 수량과 품질을 향상시킬 수 있었다. Waste nutrient solution (WNS) using non-recycled hydroponics for paprika (Capsicum annuum L.) decreased NH₄-N, but increased Mg and SO₄-S concentration as the paprika plants grew. EC and pH of WNS were maintained well at ± 5% and ± 15% levels of supplied solution, respectively. The paprika plants cultivated by WNS 2.0 dS · m?¹ showed thicker stem diameter and higher leaf area index (LAI), but there were no significant differences in the plant height, number of leaf, and leaf thickness. The paprika plants cultivated by WNS showed higher chlorophyll content as they grew. The blossom end rot (BER) of fruits did not appear in WNS treatment, but the occurrence rate of BER was 5.3% in the fruits grown by pepper Hanbang fertigation solution (1.5 dS · m?¹, N: 9.4% (A)+11% (B), P: 6%, K: 28% (A)+22% (B), Mg: 3.5%, B: 0.1%, Zn: 0.003%, Cu: 0.0005%, Mo: 0.0005%). The total yield and number of fruit were higher in WNS 2.0 dS · m?¹ treatment compared to the others. The paprika leaf grown by WNS had higher Mg and Ca contents. The K content of paprika leaf was higher in pepper Hanbang fertigation solution treatment. The fruit size of paprika classified by weight showed difference among treatments. The medium size fruits (100-150 g) were above 52% in WNS 2.0 dS · m?¹ and Hanbang fertigation solution, whereas the percentage of small (<100 g) and large (>150 g) size fruits were the highest in Hanbang fertigation solution and WNS 2.0 dS · m?¹ treatments, respectively. Therefore, for paprika plants cultivated in WNS 2.0 dS · m?¹, growth, yield and quality of fruit could be enhanced more than fertigation solution in fertigation culture.