• Applied Biological Chemistry
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• 제46권1호
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• pp.1-6
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• 2003

본 연구는 폐수 중의 질소 제거를 위한 생물학적 처리용 미생물 개발을 위한 목적으로 질소의 산화 능력이 뛰어난 균주를 분리하였다. 분리된 세균 중에서 질소 산화능과 생육 속도가 뛰어난 CH-N 균주를 선별하였으며, 생리, 생화학적 특성 조사에 의해 Bacillus sp로 추정되어 Bacillus sp. CH-N이라 명명하였다. 분리 균주는 0.5% glucose가 포함된 초기 pH가 7,0인 암모니아 및 아질산성 질소 함유 배지에서 30시간 배양 후 각각 85%와 90%의 암모니아성과 아질산성 질소의 감소율을 나타내었다. 폐수 및 생활하수에 분리 균주를 이용한 결과, 수질 속의 암모니아성 질소가 단시간에 크게 감소시키는 효과를 확인하였다. 균주를 고정시킨 담체의 질소산화 효과를 시험하고자 Bacillus sp. CH-N을 고정시킨 세라믹 담체를 이용한 결과, 배양 2일 후에는 암모니아성 질소가 전부 제거되었다.

• Environmental Engineering Research
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• 제15권2호
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• pp.71-78
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• 2010

To understand the effects of acclimation schemes on the formation of anode biofilms, different electrical performances are characterized in this study, with the roles of suspended and attached bacteria in single-chamber microbial fuel cells (MFCs). The results show that the generation of current in single-chamber MFCs is significantly affected by the development of a biofilm matrix on the anode surface containing abundant immobilized microorganisms. The long-term operation with suspended microorganisms was demonstrated to form a dense biofilm matrix that was able to reduce the activation loss in MFCs. Also, a Pt-coated anode was not favorable for the initial or long-term bacterial attachment due to its high hydrophobicity (contact angle = $124^{\circ}$), which promotes easy detachment of the biofilm from the anode surface. Maximum power ($655.0\;mW/m^2$) was obtained at a current density of $3,358.8\;mA/m^2$ in the MFCs with longer acclimation periods. It was found that a dense biofilm was able to enhance the charge transfer rates due to the complex development of a biofilm matrix anchoring the electrochemically active microorganisms together on the anode surface. Among the major components of the extracellular polymeric substance, carbohydrates ($85.7\;mg/m^2_{anode}$) and proteins ($81.0\;mg/m^2_{anode}$) in the dense anode biofilm accounted for 17 and 19%, respectively, which are greater than those in the sparse anode biofilm.

• 한국전문물리치료학회지
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• 제11권2호
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• pp.47-63
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• 2004

In most tissues, apoptosis plays a pivotal role in normal development and in regulation of cell number. Therefore inappropriate apoptosis is revealed in a variety of diseases. This study was carried out to investigate the effects of acupuncture and needle electrode electrical stimulation on the change of caspase-3, 9 and neuronal nitric oxide synthase (nNOS) immunoreactive cells in the sprague dawley rats (SD rat). In immobilized SD rats (n=5), enhanced caspase-3 and caspase-9 expression were detected in the reticular part of substantia nigra, and enhanced nNOS was detected in the dorsolateral periaqueductal gray (DL-PAG) of midbrain and the paraventricular nucleus (PVN) of the hypothalamus using immunohistochemistry. Following the immobilization, acupuncture (n=5) and needle electrode electrical stimulation (n=5, 2 Hz) was applied at H$\acute{e}$g$\breve{u}$ (LI4) acupoint of SD rats, respectively. The stress-induced enhancement in the expression of caspase-3, 9 and nNOS were The present results demonstrate that and needle electrode electrical stimulation are effective in the modulation of expression of caspase-3, 9 and nNOS induced by immobilization.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2011년도 제41회 하계 정기 학술대회 초록집
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• pp.202-202
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• 2011

The research of 3-dimensional (3-D) scaffold for tissue engineering has been widely investigated as the importance of the 3-D scaffold increased. 3-D scaffold is needed to support for cells to proliferate and maintain their biological functions. Furthermore, its architecture defines the shape of the new bone and cartilage growth. Polycaprolactone (PCL) has been one of the most promising materials for fabricating 3-D scaffold owing to its excellent mechanical property and biocompatibility. However, there are practical problems for using it, in vitro and in vivo; extracellular matrix components and nutrients cannot penetrate into the inner space of scaffold, due to its hydrophobic property, and thus cell seeding and attachment onto the inner surface remain as a challenge. Thus, the surface modification strategy of 3-D PCL scaffold is prerequisite for successful tissue engineering. Herein, we utilized a mussel-inspired approach for surface modification of 3-D PCL scaffold. Modification of 3-D PCL scaffolds was carried out by simple immersion of scaffolds into the dopamine solution and stimulated body fluid, and as a result, hydroxyapatite-immobilized 3-D PCL scaffolds were obtained. After surface modification, the wettability of 3-D PCL scaffold was considerably changed, and infiltration of the pre-osteoblastic cells into the 3-D scaffold followed by the attachment onto the surface was successfully achieved.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2012년도 제42회 동계 정기 학술대회 초록집
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• pp.195-195
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• 2012

Recently, graphene based solution-gated field-effect transistors (SGFETs) have been received a great attention in biochemical sensing applications. Graphene and reduced graphene oxide (RGO) possess various advantages such as high sensitivity, low detection limit, label-free electrical detection, and ease of fabrication due to their 2D nature and large sensing area compared to 1D nanomaterials- based nanobiosensors. Therefore, graphene or RGO -based SGFET is a good potential candidate for sensitive detection of protons (H+ ions) which can be applied as the transducer in various enzymatic or cell-based biosensing applications. However, reports on detection of H+ ions using graphene or RGO based SGFETs have been still limited. According to recent reports, clean graphene grown by CVD or exfoliation is electrochemically insensitive to changes of H+ concentration in solution because its surface does not have terminal functional groups that can sense the chemical potential change induced by varying surface charges of H+ on CVD graphene surface. In this work, we used RGO -SGFETs having oxygen-containing functional groups such as hydroxyl (OH) groups that effectively interact with H+ ions for expectation of increasing pH sensitivity. Additionally, we also investigate RGO based SGFETs for bio-sensing applications. Hydroloytic enzymes were introduced for sensing of biomolecular interaction on the surface of RGO -SGFET in which enzyme and substrate are acetylcholinesterase (AchE) and acetylcholine (Ach), respectively. The increase in H+ generated through enzymatic reaction of hydrolysis of Ach by AchE immobilized on RGO channel in SGFET could be monitored by the change in the drain-source current (Ids).

• KSBB Journal
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• 제7권1호
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• pp.15-20
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• 1992

식품첨가물 등으로 이용 되어지는 sorbitol을 $\beta$-1,2fructose oligomer를 함유하고 있는 Jerusalem artichoke를 이용하여 생산 하였으며, 이는 L-sorbose생산에 있어서 전구물질로 이용된다. Inulinase와 oxidoreductase의 동시고정화여, inulin을 와 fructose로 가수분해함과 동시에 가수분해산물을 sorbitol로 전환시켰다. Inulinase와 oxidoreductase는 chitin(5%, w/v)과 K-carrageenan(4%, w/v)으로 고정화 시켰다. 0.2% CTAB(Cetyltrimethylammonlumbromide)처리에 의해서 투과성이 향상된 Zymomonas mobilis세포내의 함유 되어있는 oxidoreductase의 activity가 향상되었다. Jerusalem artichoke juice를 acid에 의한 가수분해에 의해서 40%(v/w) juice 100ml당 53.64의 total carbohydrate yield를 얻었고, 온도 변화에 따른 가수분해시 $85^{\circ}C$에서 반응이 신속히 일어났다. Inulinase를 이용한 가수분해에 의하여 35.56g/l의 glucose와 85.32g/l의 fructose가 생성되었으며, sorbitol생성에 기질로 이용되었다. Inulinase와 Z. mobilis세포의 동시고정화 반응기에서 35.15g/l의 sorbitol을 생성하였으며, 이는 동시고정화를 하지 않았을때의 sorbitol생성농도(35.64g/l)보다 낮았다.

• Journal of Microbiology
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• 제42권2호
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• pp.126-132
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• 2004

A single chain variable fragment (scFv) specific towards B. pseudomallei exotoxin had previously been generated from an existing hybridoma cell line (6E6AF83B) and cloned into the phage display vector pComb3H. In this study, the scFv was subcloned into the pComb3X vector to facilitate the detection and purification of expressed antibodies. Detection was facilitated by the presence of a hemagglutinin (HA) tag, and purification was facilitated by the presence of a histidine tag. The culture was grown at 30$^{\circ}C$ until log phase was achieved and then induced with 1 mM IPTG in the absence of any additional carbon source. Induction was continued at 30$^{\circ}C$ for five h. The scFv was discerned by dual processes-direct enzyme-linked immunosorbent assays (ELISA), and Western blotting. When compared to E. coli strains ER2537 and HB2151, scFv expression was observed to be highest in the E. coli strain Topl0F'. The expressed scFv protein was purified via nickel-mediated affinity chromatography and results indicated that two proteins a 52 kDa protein, and a 30 kDa protein were co-purified. These antibodies, when blotted against immobilized exotoxin, exhibited significant specificity towards the exotoxin, com-pared to other B. pseudomallei antigens. Thus, these antibodies should serve as suitable reagents for future affinity purification of the exotoxin.

• 한국임상약학회지
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• 제8권1호
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• pp.29-34
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• 1998

Murine CD38 is a 42 kDa type II glycoprotein expressed on cell surface of both B and T lymphocytes. CD38 is a multifunctional enzyme that catalyzes the formation and hydrolysis of cyclic adenosine diphosphoribose (cADPR): ADP-ribosyl cyclase activity of CD38 catalyzes the formation of cADPR from NAD and cADPR hydrolase activity of CD38 catalyzes the hydrolysis of cADPR to ADP-ribose (ADPR). And also, CD38 has the catalytic activity of NAD glycohydrolase (NADase) which catalyzes the hydrolysis of catalyzes the formation and hydrolysis of cyclic adenosine diphosphoribose (cADPR): ADP-ribosyl cyclase activity of CD38 catalyzes the formation of cADPR from NAD to ADPR. In this study, we attempted to purify CD38 from mouse lymphocytes by using the immobilized anti-CD38 monoclonal antibody. The single step immuno-affinity column chromatography resulted in homogeneous purification, showing a single protein of 42 kDa on a SDS polyacrylamide gel. We have investigated the effects of various inhibitors on the enzyme activities of the purified CD38. Cibacron blue (0.5 mM) inhibited all three enzyme activities of CD38, NADase, ADP-ribosyl cyclase and cADPR hydrolase activities. ADPR (2 mM) showed inhibitory effect on both cADPR hydrolase activity and NADase, but not on ADP-ribosyl cyclase activity. However, ATP (2 mM) inhibited only cADPR hydrolase activity. $Zn^{2+}$ (1 mM) showed similar inhibitory effect as that of ADPR, but activated cyclase activity These results suggest that CD38 has three different catalytic activity domains which might be differentially regulated by their specific inhibitors.

• Journal of Microbiology and Biotechnology
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• 제13권4호
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• pp.509-516
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• 2003

A model system for the immnunochemical detection of Escherichia coli O157:H7 using a combined immunomagnetic separation (IMS) and test-strip liposome immunoassay (LIA) procedure was developed. Immunomagnetic beads coated with anti-E. coli O157 IgG antibodies were used to separate the E. coli O157 (including the H7 serotype) from culture. Immunoliposomes, whose surface was conjugated to goat anti-E. coli O157:H7 IgG and which encapsulated the marker dye, sulforhodamine B, were used as a detection label. The test strip, onto which antibodies to goat IgG were immobilized, was the immunosensor capturing immunoliposomes that did not bind to E. coli O157:H7 on the immunomagnetic bead-E. coli O157:H7 complexes. In experiments, pure cell culture suspensions of $10^5 E.$ coli O157:H7 organisms per ml produced a measurable signal inhibition, whereas a weak yet detectable signal inhibition occurred with $10^3CFU/ml$. The inhibition signals increased, when the incubation time for IMS was extended to 90 min and higher IgG-tag density (0.4mol%) was used on the liposomes. With 0.2 and 0.4mol% IgG-tagged liposomes, the IMS-LIA procedure showed more improved signal inhibitions than those of a direct (no IMS) LIA. The combined assay, which measures the instantaneous signal from immunoliposomes, can be completed within 90 min, making it significantly faster than conventional plating methods and enzyme-linked immunosorbent assay (ELISA). Accordingly, it is quite feasible to use the combined immunoassay format of IMS and dye-loaded immunoliposomes for the detection of E. coli O157:H7.

• 한국미생물·생명공학회지
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• 제28권2호
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• pp.117-123
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• 2000

P. phosphoreum의 생존과 생체발광도는 온도에 의해 많은 영향을 받는다. 냉동 저장한 세포의 경우 glycerol의 보호 작용으로 세포농도와 생균수는 측정기간 동안 일정하게 유지된 반면 생체발광도는 glycerol 첨가 직후 급속히 감소하였으며, 저장 이후에도 감소된 생체발광도가 활성화되지 못하였다. 최적 생육온도인 $20^{\circ}C$의 경우 저장 초기 세포가 성장함에 따라 세포수의 증가를 보였으나 일정 시간 이후 세포 분해 현상으로 인하여 생균수 및 세포 집락수의 감소를 나타내었으며, 생체발광도는 저장 3일 이후 소멸되었다. 이와는 대조적으로 $4^{\circ}C$에 저장한 세포의 생체발광도는 저장 10일 동안 지속되어 가장 높은 생체발광 유지도를 나타내었으나 장기간 저온 저장으로 인하여 세포가 VBNC 상태에 돌입됨에 따라 총균수와 생균수는 일정한 반면 저장 10일 이후 세포 집락수의 급격한 감소를 나타내었으며, 저장 20일 이후 간균에서 구균으로 세포 형태상의 변화를 나타내었다. 이에 따라 세포 저장 시 접종원의 농도를 달리하여 VBNC 상태와 생체발광도의 관련성을 조사한 결과 VBNC 세포가 증가할수록 생체발광도의 감소를 나타내었다. 따라서 VBNC 세포를 감소시키기 위하여 세포를 고정화하여 저장한 결과 별도의 활성제 없이 실온에서 다시 활성화되어 고정화하지 않은 세포에 비해 2.3배 높은 생체발광유지도를 나타내었으며, 저온저장에 따른 platebility 소실과 세포 응축현상이 나타나지 않았다. 이러한 결과는 세포의 고정화 방법을 이용하여 $4^{\circ}C$에서도 세포의 생존 및 생체발광 유지도를 향상시킬 수 있으며, 동결 건조법의 단점을 보완해 줄 것으로 생각된다.