• 멤브레인
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• 제8권1호
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• pp.22-28
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• 1998

혈중 글루코우즈의 농도를 측정하기 위한 진단막의 제조를 위해 다층 젤라틴 필름이 사용되었다. 혈당치를 얻기위해 glucose dehydrogenase와 diaphorase를 이용하여 다층 젤라틴 필름을 통과하는 글루코우즈의 확산속도를 측정하였다. 글루코우즈의 최대 확산속도에 미치는 코팅의 간격거리, 효소의 양, 필름의 두께, 그리고 외부 온도의 영향에 대해 조사하였다.

• Journal of Genetic Medicine
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• 제20권1호
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• pp.6-14
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• 2023

Fabry disease (FD), a rare X-linked lysosomal storage disorder, is caused by mutations in the α-galactosidase A gene gene encoding α-galactosidase A (α-Gal A). The functional deficiency of α-Gal A results in progressive accumulation of neutral glycosphingolipids, causing multi-organ damages including cardiac, renal, cerebrovascular systems. The current treatment is comprised of enzyme replacement therapy (ERT), oral pharmacological chaperone therapy and adjunctive supportive therapy. ERT has been introduced 20 years ago, changing the outcome of FD patients with proven effectiveness. However, FD patients have many unmet needs. ERT needs a life-long intravenous therapy, inefficient bio-distribution, and generation of anti-drug antibodies. Migalastat, a pharmacological chaperone, augmenting α-Gal A enzyme activity only in patients with mutations amenable to the therapy, is now available for clinical practice. Furthermore, these therapies should be initiated before the organ damage becomes irreversible. Development of novel drugs aim at improving the clinical effectiveness and convenience of therapy. Clinical trial of next generation ERT is underway. Polyethylene glycolylated enzyme has a longer half-life and potentially reduced antigenicity, compared with standard preparations with longer dosing interval. Moss-derived enzyme has a higher affinity for mannose receptors, and seems to have more efficient access to podocytes of kidney which is relatively resistant to reach by conventional ERT. Substrate reduction therapy is currently under clinical trial. Gene therapy has now been started in several clinical trials using in vivo and ex vivo technologies. Early results are emerging. Other strategic approaches at preclinical research level are stem cell-based therapy with genome editing and systemic mRNA therapy.

• Journal of Microbiology and Biotechnology
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• 제21권10호
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• pp.1057-1063
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• 2011

Herein, a novel ginsenosidase, named ginsenosidase type IV, hydrolyzing 6-O-multi-glycosides of protopanaxatriol-type ginsenosides (PPT), such as Re, R1, Rf, and Rg2, was isolated from the Aspergillus sp. 39g strain, purified, and characterized. Ginsenosidase type IV was able to hydrolyze the 6-O-${\alpha}$-L-($1{\rightarrow}2$)-rhamnoside of Re and the 6-O-${\beta}$-D-($1{\rightarrow}2$)-xyloside of R1 into ginsenoside Rg1. Subsequently, it could hydrolyze the 6-O-${\beta}$-D-glucoside of Rg1 into F1. Similarly, it was able to hydrolyze the 6-O-$_{\alpha}$-L-($1{\rightarrow}2$)-rhamnoside of Rg2 and the 6-O-${\beta}$-D-($1{\rightarrow}2$)-glucoside of Rf into Rh1, and then further hydrolyze Rh1 into its aglycone. However, ginsenosidase type IV could not hydrolyze the 3-O- or 20-O-glycosides of protopanaxadiol-type ginsenosides (PPD), such as Rb1, Rb2, Rb3, Rc, and Rd. These exhibited properties are significantly different from those of glycosidases described in Enzyme Nomenclature by the NC-IUBMB. The optimal temperature and pH for ginsenosidase type IV were $40^{\circ}C$ and 6.0, respectively. The activity of ginsenosidase type IV was slightly improved by the $Mg^{2+}$ ion, and inhibited by $Cu^{2+}$ and $Fe^{2+}$ ions. The molecular mass of the enzyme, based on SDS-PAGE, was noted as being approximately 56 kDa.

• Bulletin of the Korean Chemical Society
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• 제24권7호
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• pp.931-936
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• 2003

By the use of multi-loop thermodynamic boxes developed here by us, we show that models of enzyme catalysis (e.g., split-site model) developed in an attempt to emphasize the importance of the reactant-state destabilization and, thus, demonstrate misleading nature of the fundamentalist position which defines Pauling's transition-state stabilization as the entire and sole source of enzyme catalytic power, should be reduced to the fundamentalist formulation which completely neglects dynamical aspects of mechanism between the reactant and the transition states and dwells only on events restricted to the reactant and transition states alone, because the splitsite (and other canonical) formulations as well as fundamentalist formulations are based, in common, on equilibrium assumptions stipulated by the thermodynamic box logics. We propose to define the equilibrium assumptions as the requisite and sufficient conditions for the fundamentalist position to enjoy its primacy as central dogma, but not as sufficient conditions for its validity, because it is subjected to contradictions presented by existing data.

• Asian-Australasian Journal of Animal Sciences
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• 제27권2호
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• pp.290-301
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• 2014

The first purpose of this review is to outline some of the background information necessary to understand the mechanisms of action of fibre-degrading enzymes in non-ruminants. Secondly, the well-known and understood mechanisms are described, i) eliminating the nutrient encapsulating effect of the cell wall and ii) ameliorating viscosity problems associated with certain Non Starch Polysaccharides, particularly arabinoxylans and ${\beta}$-glucans. A third, indirect mechanism is then discussed: the activity of such enzymes in producing prebiotic oligosaccharides and promoting beneficial cecal fermentation. The literature contains a wealth of information on various non starch polysaccharide degrading enzyme (NSPase) preparations and this review aims to conclude by discussing this body of work, with reference to the above mechanisms. It is suggested that the way in which multi- versus single-component products are compared is often flawed and that some continuity should be employed in methods and terminology.

• The Plant Pathology Journal
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• 제32권3호
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• pp.173-181
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• 2016

Gene disruption by homologous recombination is widely used to investigate and analyze the function of genes in Fusarium fujikuroi, a fungus that causes bakanae disease and root rot symptoms in rice. To generate gene deletion constructs, the use of conventional cloning methods, which rely on restriction enzymes and ligases, has had limited success due to a lack of unique restriction enzyme sites. Although strategies that avoid the use of restriction enzymes have been employed to overcome this issue, these methods require complicated PCR steps or are frequently inefficient. Here, we introduce a cloning system that utilizes multi-fragment assembly by In-Fusion to generate a gene disruption construct. This method utilizes DNA fragment fusion and requires only one PCR step and one reaction for construction. Using this strategy, a gene disruption construct for Fusarium cyclin C1 (FCC1), which is associated with fumonisin B1 bio-synthesis, was successfully created and used for fungal transformation. In vivo and in vitro experiments using confirmed fcc1 mutants suggest that fumonisin production is closely related to disease symptoms exhibited by F. fujikuroi strain B14. Taken together, this multi-fragment assembly method represents a simpler and a more convenient process for targeted gene disruption in fungi.

• Mass Spectrometry Letters
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• 제11권3호
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• pp.52-58
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• 2020

Histidine phosphorylation (pHis) is increasingly recognized as an important post translational modification (PTM) in regulating cellular functions in eukaryotes. In order to clarify the role of pHis in mammalian cell signaling system, a global phosphorylation study was performed in human prostate cancer cells, PC-3M, using a TiO₂ affinity chromatography. A total number of 307 pHis sites were identified on the 268 proteins among total identified 9,924 phosphorylation sites on 3,316 proteins. In addition, 22 pHis proteins were classified in enzyme category. This report provides the first database for the study of pHis in prostate cancer cells.

• Journal of Dairy Science and Biotechnology
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• 제41권2호
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• pp.45-56
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• 2023

This review serves two purposes: first, to promote the use of improved optimization techniques in response surface methodology (RSM); and second, to enhance the optimum conditions for the fermentation of probiotics. According to research in dairy science, Lactiplantibacillus plantarum K79 is a candidate probiotic that has beneficial health effects, such as lowering blood pressure. The optimum conditions for L. plantarumK79 to produce peptides with angiotensin-converting enzyme (ACE) inhibitory activity were proposed, through modeling each of ACE inhibitory activity and pH as a function of the four factors that are skim milk concentration (%), incubation temperature (℃), incubation time (hours), and starter added amount (%). To estimate optimum conditions, the researchers employed a desirability-based multi-response optimization approach, utilizing third-order models with a nonsignificant lack of fit. The estimated optimum fermentation conditions for L. plantarum K79 were as follows: a skim milk concentration of 10.76%, an incubation temperature of 36.9℃, an incubation time of 23.76 hours, and a starter added amount of 0.098%. Under these conditions, the predicted ACE inhibitory activity was 91.047%, and the predicted pH was 4.6. These predicted values achieved the objectives of the multi-response optimization in this study.

• The Plant Pathology Journal
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• 제17권2호
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• pp.106-109
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• 2001

A multi-rapid immunofilter paper assay (multi-RIPA) system was prepared for simultaneous diagnosis of three Tobamoviruses, Cucumber green mottle mosaic virus (CGMMV), Kyuri green mottle mosaic virus (KGMMV), and Zucchini green mottle mosaic virus (ZGMMV) in cucurbitaceous crops. Each of these viruses was specifically detected by the multi-RIPA from cucumber, watermelon, zucchini, and bottle gourd inoculated with the three Tobamoviruses singly or in combination. The three viruses could infect cucumber, watermelon, and bottle gourd ; however, CGMMV could not infect zucchini as the latex-coated CGMMV antibody showed a negative reaction in the multi-RIPA of the virus-infected plant extract. When the minimum detection level of multi-RIPA was compared with that of double antibody sandwich-enzyme-linked immunosorbent assay (DAS-ELISA) using CGMMV, the latter was 10 times more sensitive than the former. The detection limit of the multi-RIPA for the purified CGMMV was 50 ng/ml. In a survey of the threeviruses in cucurbits growing in commercial fields in 1999 and 2000, CGMMV was detected in watermelon and cucumber, and ZGMMV was detected only in zucchini growing in plastic houses at the suburbs of Chonju, Korea. However, KGMMV was not found in the commercially growing cucurbit crops in our study, The results suggest that the multi-RIPA can be a simple, rapid, specific and convenient tool to detect simultaneously the Tobamoviruses.

• 한국식품영양과학회지
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• 제43권9호
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• pp.1388-1393
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• 2014

선행연구에서 얻은 alkalophilic Bacillus I-5 유래의 CGTase wild-type과 가수분해능이 강화된 mutant 효소를 활용하여 waxy rice starch로부터 아밀로펙틴 클러스터를 제조하였다. SEC-MALLS-RI 분석법으로 CGTase wild-type과 mutant 효소가 처리된 시료의 평균 분자량을 확인한 결과 10분가량의 효소반응으로 두 반응 모두 평균 분자량은 $10^4{\sim}10^5Da$으로 급격히 감소하였음을 확인하였으며, 일정 반응 시간이 경과한 이후에는 더 이상 분자량의 감소가 일어나지 않음으로 미루어 시료가 아밀로펙틴 클러스터 단위로 분해되었으며 그 분자량은 $10^4{\sim}10^5Da$ 정도임을 알 수 있다. 또한 MALDI-TOF/MS 분석을 통하여 CGTase wild-type은 다양한 종류의 cyclic 형태의 maltodextrin을 생성하고 있으며 mutant 효소는 주로 소량의 maltooligosaccharide 들을 생산함을 확인하였다.