• Applied Biological Chemistry
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• 제38권4호
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• pp.320-324
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• 1995

쥐 뇌의 microsome으로 부터 ${\Delta}^{5}-3{\beta}$-hydroxy steroid acyl 전이효소(${\Delta}^{5}-3{\beta}$-OH-SAT)를 detergent를 이용하여 용해한 뒤에 인산지방질 liposome에서 재구성하여 처리하지 않은 microsome의 ${\Delta}^{5}-3{\beta}$-OH-SAT의 활성도와 비교하였다. 용해에 사용한 detergent중에는 deoxycholic acid가 Tween 80이나 Triton X-100 보다 우수한 ${\Delta}^{5}-3{\beta}$-OH-SAT 활성도를 나타냈으며, liposome 생성에 사용한 인산지방질중에서는 난황 및 대두 phosphatidyl-choline이 가장 높은 활성도를 보였으며 그 다음이 phosphatidylethanolamine이었고 phosphatidylserine과 phosphatidic acid는 재구성하지 않은 용해된 효소보다 ${\Delta}^{5}-3{\beta}$-OH-SAT 활성도가 낮았다. 따라서 인산지방질의 head group의 4차 암모늄기나 amine기는 ${\Delta}^{5}-3{\beta}$-OH-SAT의 재구성활성을 촉진시키고 COOH기가 amine기와 함께 존재하거나 amine기가 없으면 이 효소의 활성이 저해됨을 알 수 있었다.

• 한국지하수토양환경학회지:지하수토양환경
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• 제11권5호
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• pp.43-50
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• 2006

Bioreporter 균주는 복잡한 환경매체의 특정 오염원 탐지를 위해 유용하게 사용되고 있다. 특히 발광 유전자 재조합 균주는 민감하고 배경에 의한 영향을 받지 않는 장점이 있다. 사용한 유전자 재조합 균주(Pseudomonas putida mt-2 KG1206)는 TOL 플라즈미드와 pUCD615 벡터에 $P_{m}\;promoter$가 삽입된 재조합 플라즈미드를 함유하고 있으며, 톨루엔 계열 및 중요 분해산물에 대해 분해와 함께 발광을 생산하는 특성을 갖고 있다. 본 연구에서는 균주 동결 및 동결건조 준비 및 적용과정에 필요한 다양한 조건들을 조사하여, 향후 환경매체에 적용하기 위한 최적 방법에 대한 프로토콜을 작성하였다. 조사한 최적 조건들은 다음과 같다. 동결보호시약(24% sucrose), 동결건조 시간(12시간), 균주 농도($OD_{600}=0.6$), 동결균주 활성회복($35^{\circ}C$에서 빠르게 해동), 동결건조 균주 활성회복(LB배지에 $3{\sim}6$시간 노출), 현장 운반 조건(활성 회복 후 $20^{\circ}C$ 정도의 실온). 본 연구 결과는 재조합 균주 환경 적용을 위해 필요한 균주 동결 및 동결 건조에 대한 중요한 자료들을 제시하고 있다.

• Journal of Microbiology and Biotechnology
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• 제20권4호
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• pp.727-731
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• 2010

The gene APE0743 encoding the superoxide dismutase (ApSOD) of a hyperthermophilic archaeon Aeropyrum pernix K1 was cloned and overexpressed as a GST fusion protein at a high level in Escherichia coli. The expressed protein was simply purified by the process of glutathione affinity chromatography and thrombin treatment. The ApSOD was a homodimer of 25 kDa subunits and a cambialistic SOD, which was active with either Fe(II) or Mn(II) as a cofactor. The ApSOD was highly stable against high temperature. This thermostable ApSOD is expected to be applicable as a useful biocatalyst for medicine and bioindustrial processes.

• BMB Reports
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• 제53권9호
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• pp.466-471
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• 2020

Several humanized mouse models are being used to study humanspecific immune responses and diseases. However, the pivotal needs of fetal tissues for the humanized mice model have been huddled because of the demand for ethical and medical approval. Thus, we have verified the hematopoietic and immunomodulatory function of HepaRG and developed a new and easy humanized mouse model to replace the use of fetal liver tissue. HepaRG co-transplanted Hu-NSG mice significantly increased CD45+ lymphocytes and CD19+ B cells and CD3+ T cells than normal Hu-NSG, suggesting enhanced reconstitution of the human immune system. These results have improved the applicability of humanized mice by developing new models easily accessible.

• BMB Reports
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• 제35권3호
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• pp.255-261
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• 2002

GTP cyclohydrolase I (E.C. 3.5.4.16) is a homodecameric protein that catalyzes the conversion of GTP to 7,8-dihydroneopterin triphosphate (H2NTP), the initial step in the biosynthesis of pteridines. It was proposed that the enzyme complex could be composed of a dimer of two pentamers, or a pentamer of tightly associated dimers; then the active site of the enzyme was located at the interface of three monomers (Nar et al. 1995a, b). Using mutant enzymes that were made by site-directed mutagenesis, we showed that a decamer of GTP cyclohydrolase I should be composed of a pentamer of five dimers, and that the active site is located between dimers, as analyzed by a series of size exclusion chromatography and the reconstitution experiment. We also show that the residues Lys 136, Arg139, and Glu152 are of particular importance for the oligomerization of the enzyme complex from five dimers to a decamer.

• 한국축산식품학회지
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• 제38권6호
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• pp.1273-1285
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• 2018

The aim of this study is to determine the some physicochemical (proximate composition, pH and $a_w$ values) and reconstitution (wettability, dispersibility and solubility index (SI)) properties of spray-dried bone broth powder (BBP) along with the effects of the addition of different carrier agents. Subsequently, the powdered products were stored to determine the storage stability (pH, lipid oxidation, color, browning index) for 3 mon at $-18^{\circ}C$. For this purpose, firstly marrow-containing bones (Os femur and Os humerus) have been boiled to get the bone broth. Three different emulsions were respectively prepared (1) 20% maltodextrin (MD) added BBP, (2) 20% whey powder isolate (WPI) added BBP and (3) 10% MD and 10% WPI added BBP and the emulsions were dried using a spray-dryer with $185^{\circ}C$ inlet and $95^{\circ}C$ outlet temperature. The proximate composition and reconstitution properties of BBP were found statistically different (p<0.05) depending on the use of different carrier agents. MD added BBP showed uniform and smooth morphology. The color, browning index, thiobarbituric acid reactive substances (TBARS) and pH values of BBP significantly changed (p<0.05) during storage. In conclusion, both the results of physicochemical and storage period analyses showed that the most suitable encapsulation material in the production of the bone broth powder is MD.

• BMB Reports
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• 제54권3호
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• pp.157-163
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• 2021

The transient interactions between cellular components, particularly on membrane surfaces, are critical in the proper function of many biochemical reactions. For example, many signaling pathways involve dimerization, oligomerization, or other types of clustering of signaling proteins as a key step in the signaling cascade. However, it is often experimentally challenging to directly observe and characterize the molecular mechanisms such interactions-the greatest difficulty lies in the fact that living cells have an unknown number of background processes that may or may not participate in the molecular process of interest, and as a consequence, it is usually impossible to definitively correlate an observation to a well-defined cellular mechanism. One of the experimental methods that can quantitatively capture these interactions is through membrane reconstitution, whereby a lipid bilayer is fabricated to mimic the membrane environment, and the biological components of interest are systematically introduced, without unknown background processes. This configuration allows the extensive use of fluorescence techniques, particularly fluorescence fluctuation spectroscopy and single-molecule fluorescence microscopy. In this review, we describe how the equilibrium diffusion of two proteins, K-Ras4B and the PH domain of Bruton's tyrosine kinase (Btk), on fluid lipid membranes can be used to determine the kinetics of homodimerization reactions.

• Clinical and Experimental Pediatrics
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• 제56권1호
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• pp.26-31
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• 2013

Purpose: Lymphocyte subset recovery is an important factor that determines the success of hematopoietic stem cell transplantation (HSCT). Temporal differences in the recovery of lymphocyte subsets and the factors influencing this recovery are important variables that affect a patient's posttransplant immune reconstitution, and therefore require investigation. Methods: The time taken to achieve lymphocyte subset recovery and the factors influencing this recovery were investigated in 59 children who had undergone HSCT at the Department of Pediatrics, The Catholic University of Korea Seoul St. Mary's Hospital, and who had an uneventful follow-up period of at least 1 year. Analyses were carried out at 3 and 12 months post-transplant. An additional study was performed 1 month post-transplant to evaluate natural killer (NK) cell recovery. The impact of pre- and post-transplant variables, including diagnosis of Epstein-Barr virus (EBV) DNAemia posttransplant, on lymphocyte recovery was evaluated. Results: The lymphocyte subsets recovered in the following order: NK cells, cytotoxic T cells, B cells, and helper T cells. At 1 month post-transplant, acute graft-versus-host disease was found to contribute significantly to the delay of $CD16^+/56^+$ cell recovery. Younger patients showed delayed recovery of both $CD3^+/CD8^+$ and $CD19^+$ cells. EBV DNAemia had a deleterious impact on the recovery of both $CD3^+$ and $CD3^+/CD4^+$ lymphocytes at 1 year post-transplant. Conclusion: In our pediatric allogeneic HSCT cohort, helper T cells were the last subset to recover. Younger age and EBV DNAemia had a negative impact on the post-transplant recovery of T cells and B cells.

• 한국식품과학회지
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• 제32권4호
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• pp.881-888
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• 2000

본 연구에서는 냉동건조에 의한 생균수 감소를 줄이기 위하여 첨가된 냉동건조보호제(Freeze Drying Protectant : FDP)가 우유 또는 난백분말로 만든 젖산균 발효식품의 품질(관능적 특성, 휘발성 향기 성분, 물성)에 미치는 영향을 조사하였다. 우유시료에는 0.2%의 Tween 80과 1%의 ascorbate를 각각 첨가하고, 난백분말시료에는 3%의 raffinose와 1%의 ascorbate를 각각 첨가하여 다음과 같은 결과를 얻었다. (1) 냉동건조시키지 않은 시료의 관능적 특성은 표준시료(우유시료)와 비교하여 ascorbate 첨가 우유시료는 관능적 특성이 다소 우수하였으나, 난백분말시료 또는 FDP 첨가 난백분말시료는 관능적 특성이 다소 저조하였다. (2) 냉동건조 후 복원시킨 시료의 관능성은 표준시료(우유시료)와 다른 5종의 시료 사이에 차이가 없었다. (3) Ascorbate 첨가 우유시료는 냉동건조/복원에 의하여 관능적 특성이 저하되었고, ascorbate 첨가 난백분말시료는 냉동건조/복원에 의하여 관능적 특성이 변화하지 않았다. (4) 휘발성 향기 성분의 분석 결과를 보면, 냉동건조에 의하여 모든 휘발성 향기 성분 함량이 감소하였으며, 우유시료군과 난백분말시료군 사이에는 회수율이 다소 다른 pattern을 보였다. (5) 휘발성 향기성분의 종류에 따라 다르지만 우유시료군과 난백분말 시료군의 휘발성 향기 성분 함량은 냉동건조 전에는 차이가 비교적 컸으나, 냉동건조/복원한 후에는 차이가 비교적 작았다. (6) 냉동건조에 의하여 우유시료군의 물성은 크게 변화하였으나, 난백분말시료군의 물성변화는 비교적 작았다.

• 산업식품공학
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• 제14권1호
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• pp.54-59
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• 2010

본 연구에서는 영양과 건강, 그리고 편의성까지 고려한 즉석식품 형태의 쌀미음을 제조하였다. 제조된 쌀미음 분말의 영양성분 구성비를 일반성분 분석을 통하여 분석 하였고, 과립공정에 따른 재수화능의 향상을 연구하였다. 과립분말은 혼합분말보다 지방함량과 단백질 함량이 각각 0.9, 1.9% 높게 나타난 반면, 탄수화물 함량은 상대적으로 3.2% 낮게 나타났다. 과립분말의 단백질 조성은 71.1-118.5 g으로 1일 권장 단백질 섭취량 45-55 g보다 많았는데, 이는 일반적인 환자식이 지니는 공통적인 특성이다. 분말 입자의 침강 속도를 보여주는 분산성의 경우 과립분말(93.7%)은 혼합분말(77.0%) 보다 높게 나타났고, 재수화에 걸리는 시간도 과립분말(122.3 초)이 혼합분말(305.3 초)보다 빨랐으며 이들 결과는 과립공정이 분말의 재수화능을 향상시켜 주었음을 보여준다. 호화쌀가루를 이용한 쌀미음은 기존의 우유 단백질 위주의 환자식을 곡류로 대체하며 영양강화 쌀 가공제품의 개발에 의미가 있다.