High-level extracellular production and characterization of Candida antarctica lipase B in Pichia pastoris

Eom, G.T.,Lee, S.H.,Song, B.K.,Chung, K.W.,Kim, Y.W.,Song, J.K. Society for Bioscience and Bioengineering, Japan ; 2013 Journal of bioscience and bioengineering Vol.116 No.2

The gene encoding lipase B from Candida antarctica (CalB) was expressed in Pichia pastoris after it was synthesized by the recursive PCR and cloned into the Pichia expression plasmid, pPICZαA. The CalB was successfully secreted in the recombinant P. pastoris strain X-33 with an apparent molecular weight of 34 kDa. For 140 h flask culture, the dry cell weight and the extracellular lipase activity reached at 5.4 g/l and 57.9 U/l toward p-nitrophenyl palmitate, respectively. When we performed the fed-batch fermentation using a methanol feeding strategy for 110 h, the dry cell weight and the extracellular lipase activity were increased to 135.7 g/l and 11,900 U/l; the CalB protein concentration was 1.18 g/l of culture supernatant. The characteristics of CalB recovered from the P. pastoris culture were compared with the commercial form of CalB produced in Aspergillus oryzae. The kinetic constants and specific activity, the effects of activity and stability on temperature and pH, the glycosylation extent, the degree of immobilization on macroporous resin and the yield of esterification reaction between oleic acid and n-butanol were almost identical to each other. Therefore, we successfully proved that the Pichia-based expression system for CalB in this study was industrially promising compared with one of the most efficient production systems.

Clinical Outcome of Autologous Hematopoietic Cell Transplantation in Adult Patients with Acute Myeloid Leukemia: Who May Benefit from Autologous Hematopoietic Cell Transplantation?

Yoon, J.H.,Kim, H.J.,Park, S.S.,Jeon, Y.W.,Lee, S.E.,Cho, B.S.,Eom, K.S.,Kim, Y.J.,Lee, S.,Min, C.K.,Cho, S.G.,Kim, D.W.,Lee, J.W.,Min, W.S. AMERICAN SOCIETY FOR BLOOD AND MARROW 2017 BIOLOGY OF BLOOD AND MARROW TRANSPLANTATION Vol.23 No.4

The role of autologous hematopoietic cell transplantation (auto-HCT) for postremission therapy of acute myeloid leukemia is yet to be elucidated. We retrospectively analyzed 240 patients treated with auto-HCT in first remission. All patients were treated with standard induction chemotherapy, and CD34<SUP>+</SUP>@?stem cells were collected at each cycle of consolidation. Stem cells were infused after total body irradiation (1200 cGy), cytarabine (9 g/m<SUP>2</SUP>), and melphalan (100@?mg/m<SUP>2</SUP>). Estimated 5-year overall survival, disease-free survival (DFS), cumulative incidence of relapse (CIR), and nonrelapse mortality were 58.4%, 55.3%, 38.8%, and 5.9%, respectively. We identified that poor-risk karyotype showed very poor outcome after auto-HCT, and then analyzed 85 patients with good to intermediate-risk molecular cytogenetics with available molecular study results and markers for minimal residual disease (MRD) such as WT1 and core-binding factor (CBF) associated MRD (ie, AML1/ETO and CBFβ/MYH11). Our data identified that old age, pre-HCT markers for MRD, and high post-HCT WT1, high dose of CD34<SUP>+</SUP>@?stem cell (≥4.5 x 10<SUP>6</SUP>/kg) infusion, and c-kit or FLT3-ITD mutations were associated with higher relapse rate and poor DFS. Using pre-HCT parameters, except for post-HCT WT1, multivariate analysis revealed that patients with young age (<40 years old), no adverse mutations, and limited dose of CD34<SUP>+</SUP>@?stem cells might be good candidate for auto-HCT (3-year DFS and CIR were 83.4% and 16.6%, respectively). Young patients with good- to intermediate-risk molecular cytogenetics may benefit from auto-HCT if stem cell dose is limited.

Stabilization/solidification of mercury-contaminated waste ash using calcium sodium phosphate (CNP) and magnesium potassium phosphate (MKP) processes

Cho, J.H.,Eom, Y.,Lee, T.G. Elsevier Scientific Pub. Co 2014 Journal of hazardous materials Vol.278 No.-

This study examined the stabilization and solidification (S/S) of mercury (Hg)-contaminated waste ash generated from an industrial waste incinerator using chemically bonded phosphate ceramic (CBPC) technology. A magnesium potassium phosphate (MKP; MgKPO<SUB>4</SUB>.6H<SUB>2</SUB>O) ceramic, fabricated from MgO and KH<SUB>2</SUB>PO<SUB>4</SUB>, and a calcium sodium phosphate (CNP; CaNaPO<SUB>4</SUB>) ceramic, fabricated from CaO and Na<SUB>2</SUB>HPO<SUB>4</SUB>, were used as solidification binders in the CBPC process, and Na<SUB>2</SUB>S or FeS was added to each solidification binder to stabilize the Hg-contaminated waste ash. The S/S processes were conducted under various operating conditions (based on the solidification binder and stabilization reagent, stabilization reagent dosage, and waste loading ratio), and the performance characteristics of the S/S sample under each operating condition were compared, including the Hg leaching value and compressive strength. The Hg leaching value of untreated Hg-contaminated waste ash was 231.3μg/L, whereas the S/S samples treated using the MKP and CNP processes exhibited Hg leaching values below the universal treatment standard (UTS) limit (25μg/L). Although the compressive strengths of the S/S samples decreased as the sulfide dosage and waste loading ratio were increased, most of the S/S samples fabricated by the MKP and CNP processes exhibited good mechanical properties.

화력발전소의 대기오염물질 배출계수 산정 연구

김대곤,엄윤성,홍지형,이석조,석광설,이대균,이은정,방선애 한국대기환경학회 2004 한국대기환경학회지 Vol.20 No.3

The main purpose of this study was to characterize the air pollutants emission factors in electric power plant (EPP) using fossil fuels. The electric power plant is a major air pollution source, thus knowing the emission characteristics of electric power plant is very important to develop a control strategy. The major air pollutants of concern from EPP stacks are particulate matter (PM), sulfur oxides (SOx), nitrogen oxides (NOx), carbon monoxide (CO) and heavy metals. Throughout the study. the following results arc estimated: - PM : 8.671E-05∼8.724E+01 PM emission (kg) per fuel burned (ton) - SOx: 4.749E-04∼7.877E+01 SOx emission (kg) per fuel burned (ton) - NOx : 1.578E-02∼9.857E+00 NOx emission (kg) per fuel burned (ton) - CO : 3.800E-04∼1.291E+00 CO emission (kg) per fuel burned (ton) - Hg : 1.220E+01∼3.108E+02 Hg emission (mg) per fuel burned (ton) From the statistical analysis by Wilcoxon signed ranks test between the emission factors of ours and U.S. EPA's. we can yielded that: p > 0.05.

장애인 주간보호시설의 비대면 체육수업 운영 경험에 관한 연구

노승덕,이근호,박철승 한국특수체육학회 2022 한국특수체육학회지 Vol.30 No.2

The purpose of this study is to understand the improvement factors of non-face-to-face physical education classes in day care center used by adults with developmental disabilities in the COVID-19 era. Accordingly, in-depth interviews were conducted with 5 sports instructors for the disabled who are working at the G sports association for the disabled. Data collection was conducted from August 2021 to February 2022, and a triangulation method such as content review of research participants and expert meetings was implemented to secure the trustworthiness of the research. For the analysis of the collected data, the topics were selected as ‘problems of non-face-to-face physical education classes’, ‘contents of online classes in day care facilities for the disabled’, and ‘plans to improve online classes’ through an inductive content analysis method. The results of this study are as follows. First, the problems of non-face-to-face physical education classes were difficulty in online class preparation process, difficulty in online class process, and poor exercise environment in daycare center for the disabled. Second, the contents of online physical education classes at daycare center for the disabled were customized classes through program transformation and online cooperative classes with daycare center managers. Third, It was found that expansion of equipment and facilities for online classes, preparation of sports instructors for the disabled and manpower support system, establishment of an online physical education system for the disabled were found to be necessary to improve online physical education classes. Based on these results, in order to activate non-face-to-face physical education classes in daycare center for the disabled, improvement of the online physical education class environment, capability development of sports instructors for the disabled and cooperation with the manger in charge of daycare center, and preparation of an online physical education system for the disabled is presumed to be necessary. 본 연구는 코로나19 상황에서 성인 발달장애인들이 이용하는 주간보호시설에서의 비대면 체육수업의 개선 요인을 이해하는데 그 목적이 있다. 이를 위해 G장애인체육회에서 근무하고 있는 장애인스포츠지도사 5명을 대상으로 심층면담을 실시하였다. 자료 수집은 2021년 8월부터 2022년 2월까지 진행하였으며, 연구의 진실성을 확보하기 위해 연구 참여자의 내용검토, 전문가회의 등 삼각검증법을 시행하였다. 수집된 자료의 분석은 귀납적 내용분석 방법을 통해 ‘비대면 체육 수업의 문제점’, ‘장애인주간보호시설에서 온라인 수업내용’, ‘온라인 수업 개선방안’으로 주제를 선정하였다. 연구 결과는 첫째, 비대면 체육 수업의 문제점은 ‘온라인 수업 준비과정의 어려움’, ‘온라인 수업 진행과정 어려움’, ‘장애인주간보호시설의 열악한 운동환경’으로 나타났다. 둘째, 장애인주간보호시설에서 온라인 체육수업 내용은 ‘프로그램 변형을 통한 맞춤형 수업’, ‘시설 담당자와 온라인 협력수업’으로 나타났다. 셋째, 온라인 체육수업 개선방안으로 ‘온라인 수업을 위한 장비 및 시설 확충’, ‘지도자의 역량 및 인력지원 체계 마련’, ‘장애인을 위한 온라인 시스템 구축’이 필요한 것으로 나타났다. 이 결과로 보아 장애인 주간보호시설의 비대면 체육수업을 활성화를 위해서는 온라인 체육수업 환경의 개선, 지도자 역량 및 주간보호시설 담당자와의 협력, 장애인을 위한 온라인 체육수업 시스템이 마련되어야 할 것으로 사료된다.

Overcoming various comorbidities by G-CSF-primed unmanipulated BM SCT in adult patients with AML

Kim, H-J,Min, W-S,Cho, B-S,Eom, K-S,Kim, S-Y,Kim, Y-J,Lee, S,Min, C-K,Cho, S-G,Lee, J-W,Kim, C-C Macmillan Publishers Limited 2009 BONE MARROW TRANSPLANTATION -BASINGSTOKE- Vol.44 No.6

We examined whether the use of G-CSF-primed unmanipulated bone marrow (pBM) permits successful transplantation in patients with adverse comorbid pretransplantation conditions. A total of 33 patients at variable risk of AML undergoing allogeneic SCT from an HLA-identical sibling participated. The patients suffered from a variety of treatment-related sequelae before SCT and many cases also featured the presence of major organ dysfunction. The median follow-up duration of patients who were among the overall survivors was 58 months (range, 11−125 months). The median number of CD34<SUP>+</SUP> cells infused was 3.5 × 10<SUP>6</SUP> per kg (range, 0.8−15.6 × 10<SUP>6</SUP> per kg). The median number of days for recovery of ANC and platelet count without transfusion was 13 (range, 6−22) and 18 (range, 11−29) days, respectively. Four patients (12%) died due to nonrelapse causes and only one patient developed the same course of infectious complication as before SCT. No particular finding in the context of CMV infection or other post transplant complication was observed. The estimated 10-year overall survival rate was 68%. Our results suggest that the use of pBM allows successful engraftment without increasing complications in patients with various comorbidities before transplantation.

중학교 성교육 지침과 실태 비교

곽예림,권경미,김유리,신지연,엄영아,오가영,윤미혜,윤예림,정자혜,최은아 이화여자대학교 간호과학대학 2010 이화간호학회지 Vol.- No.44

Nowadays, juvenile's sexual-related delinquency is increasing and their immature sexual behavior is on issue with this recent phenomenon. That is the reason people realize offering right sex education for them is important. So, we are going to study for trying to make an improvement in sex education field based on the fact that is practicing now at school. We will reference "소중한 성 바로알기" in this study which is published in 2008 for schoolteacher to compensate some limitations of former education material. This study was done from June 15 to July 20, 2009(Total : 36days) and aimed for surveying senior year students of middle school located in Seoul, S. Korea. We sampled 248 students by random from 8 classes of 4 schools. We analyzed final 209 samples(the other 39 samples were omitted for inappropriate response and return.) using SPSS WIN 12.0. The questionnaire includes 4 parts of relationship and psychosexual, physical growth and sexual health, social environment and sexual equality, and healthy family with marriage and it consists of 31 questions. We made this questionnaire base on the reference as we stated above. From this study we found the part of healthy family with marriage has not enough contents compared with the other 3 parts. And also we could see students don't feel satisfaction with one-off, one-sided sex education, and the education which is performed with only one single method. Therefore we make a judgement that middle school students have their needs toward sex education at school and those are considered as regular education by well-trained education provider, and various means for education such as case study, discussion, or role play. As well we think sex education for middle school students need to offer more about sexual equality and also alert the increasing possibilities of sexual commit in cyber space to adolescent.

Alleviation of temperature-sensitive secretion defect of Pseudomonas fluorescens ATP-binding cassette (ABC) transporter, TliDEF, by a change of single amino acid in the ABC protein, TliD

Eom, G.T.,Oh, J.Y.,Park, J.H.,Lim, H.J.,Lee, S.J.,Kim, E.Y.,Choi, J.E.,Jegal, J.,Song, B.K.,Yu, J.H.,Song, J.K. Society for Bioscience and Bioengineering, Japan ; 2016 Journal of bioscience and bioengineering Vol.122 No.3

<P>An ABC transporter, TliDEF, from Pseudomonas fluorescens SIK W1, mediates the secretion of its cognate lipase, TliA, in a temperature-dependent secretion manner; the TliDEF-mediated secretion of TliA was impossible at the temperatures over 33 degrees C. To isolate a mutant TliDEF capable of secreting TliA at 35 degrees C, the mutagenesis of ABC protein (TliD) was performed. The mutated tliD library where a random point mutation was introduced by error-prone PCR was coexpressed with the wild-type WE, tliF and tliA in Escherichia con. Among approximately 10,000 colonies of the tliD library, we selected one colony that formed transparent halo on LB-tributyrin plates at 35 degrees C. At the growth temperature of 35 degrees C, the selected mutant TliD showed 1.75 U/ml of the extracellular lipase activity, while the wild-type TliDEF did not show any detectable lipase activity in the culture supernatant of E. coli. Moreover, the mutant TliD also showed higher level of TliA secretion than the wild-type TliDEF at other culture temperatures, 20 degrees C, 25 degrees C and 30 degrees C. The mutant TliD had a single amino acid change (Ser287Pro) in the predicted transmembrane region in the membrane domain of TliD, implying that the corresponding region of TliD was important for causing the temperature-dependent secretion of TliDEF. These results suggested that the property of ABC transporter could be changed by the change of amino acid in the ABC protein. (C) 2016, The Society for Biotechnology, Japan. All rights reserved.</P>

Secretory production of enzymatically active endo-β-1,4-mannanase from Bacillus subtilis by ABC exporter in Escherichia coli

Eom, G.T.,Oh, J.Y.,Park, J.H.,Jegal, J.,Song, J.K. Elsevier Applied Science 2016 PROCESS BIOCHEMISTRY Vol.51 No.8

To secrete the endo-β-1,4-mannanase of Bacillus subtilis WL7 in Escherichia coli, the ABC exporter (TliDEF) and the C-terminal signal sequences of lipase (TliA) and metalloprotease (PrtA) were used. Six C-terminal fragments of TliA (TliAC1, TliAC2 and TliAC3) and PrtA (PrtAC1, PrtAC2 and PrtAC3) were amplified by PCR, respectively, and were fused to the mature mannanase gene lacking its original N-terminal signal sequence. When coexpressed with TliDEF exporter gene in E. coli, all the fusion mannanases were successfully secreted to the culture supernatant as an enzymatically active form by TliDEF exporter in E. coli. However, the unspecific cell leakage occurred in the fusion mannanases containing TliA-derived signal sequences. Among the fusion mannanases containing the signal sequences of PrtA, the mannanase with the shortest signal sequence fragment (Man-PrtAC3) exhibited the highest secretion level (4.65mg/L). Moreover, the specific activity, the effect of temperature and pH on activity and stability were almost identical to the wild-type mannanase, indicating that the fused signal sequence of PrtA did not affect the enzymatic properties of mannanase. It was concluded that the efficient secretion of enzymatically active mannanase from Bacillus subtilis WL-7 was achieved by using PrtAC3 as the most adequate signal sequence fragment.

Effects of Pt loading in the anode on the durability of a membrane-electrode assembly for polymer electrolyte membrane fuel cells during startup/shutdown cycling

Eom, K.,Kim, G.,Cho, E.,Jang, J.H.,Kim, H.J.,Yoo, S.J.,Kim, S.K.,Hong, B.K. Pergamon Press ; Elsevier Science Ltd 2012 International journal of hydrogen energy Vol.37 No.23

A polymer electrolyte membrane fuel cell (PEMFC) stack of a fuel cell vehicle (FCV) is inevitably exposed to reverse current conditions, which are formed by the oxygen reduction reaction (ORR) induced at the anode with a hydrogen/air boundary during startup/shutdown processes. With an increase in the reverse current, the degradation rate of the cathode that experiences a highly corrosive condition (locally high potential) increases. In this work, the anode Pt loading is decreased from 0.4 to 0.1 mg cm<SUP>-2</SUP> to decrease the reverse current. The decrease in the anode Pt loading is found to decrease the hydrogen oxidation rates (HOR) during normal operation, but this loading decrease barely affected the cell performance. However, a decrease in the anode Pt loading can significantly decrease the reverse current, leading to a diminished cathode degradation rate during startup/shutdown cycling. It is revealed by slow decreases in the cell performance (i-V curves) and electrochemical active surface area (EAS), and a slow increase in the charge-transfer resistance (R<SUB>ct</SUB>), which can be attributed to corrosion of the carbon support and dissolution/migration/agglomeration of the platinum catalyst.