• BMB Reports
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• 제51권4호
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• pp.163-164
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• 2018

CONSTANS (CO) induces the expression of FLOWERING LOCUS T (FT) in the photoperiodic pathway, and thereby regulates the seasonal timing of flowering. CO expression is induced and CO protein is stabilized by FLAVIN-BINDING KELCH REPEAT F-BOX PROTEIN 1 (FKF1) in the late afternoon, while CO is degraded by CONSTITUTIVE PHOTOMORPHOGENIC 1 (COP1) during the night. These regulatory cascades were thought to act independently. In our study, we investigated the relationship between FKF1 and COP1 in the regulation of CO stability in response to ambient light conditions. A genetic analysis revealed that FKF1 acts as a direct upstream negative regulator of COP1, in which cop1 mutation is epistatic to fkf1 mutation in the photoperiodic regulation of flowering. COP1 activity requires the formation of a hetero-tetramer with SUPPRESSOR OF PHYA-105 (SPA1), [$(COP1)_2(SPA1)_2$]. Light-activated FKF1 has an increased binding capacity for COP1, forming a FKF1-COP1 hetero-dimer, and inhibiting COP1 homo-dimerization at its coiled-coil (CC) domain. Mutations in the CC domain result in poor COP1 dimerization and misregulation of photoperiodic floral induction. We propose that FKF1 represses COP1 activity by inhibiting COP1 dimerization in the late afternoon under long-day conditions, resulting in early flowering.

• Applied Biological Chemistry
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• 제16권1호
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• pp.31-40
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• 1973

Azotobacter vinelandi의 세포추출물(細胞抽出物)들이 Shethna flavoprotein의 free radical 형(型)으로의 전자전달기구(電子傳達機構)에 관하여 연구(硏究)하였다. 전자전달(電子傳達)에 관여도(關與度)가 높은 단백질(蛋白質)로 황색형광성단백질(黃色榮光性蛋白質)(protein I)과 갈색단백질(褐色蛋白質)(protein II)을 정제(精製)하였고 이들은 $N_2$ 기압하(氣壓下) 또는 aceton-dry ice 동결(凍結)저장하에서도 쉽게 실활(失活)되었고 반응속도(反應速度) 역시 너무 완만하여 생체내반응(生體內反應)이기에는 의문점을 보였다. 한편 세포추출물중(細胞抽出物中)의 FMN은 NADH에 의(依)하여 환원이 쉽게 이루어졌으며, 환원형 $FMNH_2$는 비효소적(非酵素的)으로 Shethna flavoprotein 의 free radical 을 형성(形成)시켰으며 , 효소적반응속도(酵素的反應速度)보다 15배(倍)의 높은 반응속도(反應速度)를 보였다. 비록 FMN이 생체내(生體內)에서 타(他)단백질과 비결합형(非結合型)으로 존재(存在)하지 않는다해도 FMN의 전자전달체(電子電達體)의 가능성(可能性)을 제시(提示)하였다.

• Molecules and Cells
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• 제42권10호
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• pp.693-701
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• 2019

Plants monitor changes in day length to coordinate their flowering time with appropriate seasons. In Arabidopsis, the diel and seasonal regulation of CONSTANS (CO) protein stability is crucial for the induction of FLOWERING LOCUS T (FT) gene in long days. FLAVIN-BINDING, KELCH REPEAT, F-BOX 1 (FKF1) and ZEITLUPE (ZTL) proteins control the shape of CO expression profile antagonistically, although regulation mechanisms remain unknown. In this study, we show that GIGANTEA (GI) protein modulates the stability and nuclear function of FKF1, which is closely related to the stabilization of CO in the afternoon of long days. The abundance of FKF1 protein is decreased by the gi mutation, but increased by GI overexpression throughout the day. Unlike the previous report, the translocation of FKF1 to the nucleus was not prevented by ZTL overexpression. In addition, the FKF1-ZTL complex formation is higher in the nucleus than in the cytosol. GI interacts with ZTL in the nucleus, implicating the attenuation of ZTL activity by the GI binding and, in turn, the sequestration of FKF1 from ZTL in the nucleus. We also found that the CO-ZTL complex presents in the nucleus, and CO protein abundance is largely reduced in the afternoon by ZTL overexpression, indicating that ZTL promotes CO degradation by capturing FKF1 in the nucleus under these conditions. Collectively, our findings suggest that GI plays a pivotal role in CO stability for the precise control of flowering by coordinating balanced functional properties of FKF1 and ZTL.

• International Journal of Industrial Entomology and Biomaterials
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• 제43권1호
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• pp.29-36
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• 2021

Silkworms have long been bred with human history to produce silk. It has been with humans for longer than other industrial insects, and the silkworm variety has been continuously improved. Silkworms have been developed into the optimal form for producing high quality silk and pupae. Recently, the production of transgenic silkworms has further expanded the possibility of industrial value of silkworms. Kynurenine 3-monooxygenase (KMO), which is a flavin enzyme, is known for its involvement in ommochrome pigment synthesis. In the field of mammals, including humans, previous studies have revealed the function and role of KMO, which is an important enzyme for various immune responses and cell protection. However, in the case of insects, the function of KMO has only been studied to be involved in the formation of pigment, and accordingly, KMO is used exclusively on screening for generation of transgenic insects as a marker. In this study, using KMO-edited silkworms, it was intended to discover the novel functions and roles of KMO in silkworms by identifying changes in the expression of various genes associated with stress and growth. The changes were observed in expressions of genes regulating on stresses to survive and those on ecdysteroid hormone between wild-type (WT) silkworms and kmo mutant silkworms. The loss of KMO, in particular, decreased the expression of the shadow (sad) gene, one of the Halloween genes in the synthesis of ecdysteroid. In conclusion, these results suggest that silkworm KMO is responsible for potential functions regarding stress response and ecdysteroid synthesis.

• Journal of Microbiology and Biotechnology
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• 제32권3호
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• pp.287-293
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• 2022

The hydroxylation of methane (CH₄) is crucial to the field of environmental microbiology, owing to the heat capacity of methane, which is much higher than that of carbon dioxide (CO₂). Soluble methane monooxygenase (sMMO), a member of the bacterial multicomponent monooxygenase (BMM) superfamily, is essential for the hydroxylation of specific substrates, including hydroxylase (MMOH), regulatory component (MMOB), and reductase (MMOR). The diiron active site positioned in the MMOH α-subunit is reduced through the interaction of MMOR in the catalytic cycle. The electron transfer pathway, however, is not yet fully understood due to the absence of complex structures with reductases. A type II methanotroph, Methylosinus sporium 5, successfully expressed sMMO and hydroxylase, which were purified for the study of the mechanisms. Studies on the MMOH-MMOB interaction have demonstrated that Tyr76 and Trp78 induce hydrophobic interactions through π-π stacking. Structural analysis and sequencing of the ferredoxin domain in MMOR (MMOR-Fd) suggested that Tyr93 and Tyr95 could be key residues for electron transfer. Mutational studies of these residues have shown that the concentrations of flavin adenine dinucleotide (FAD) and iron ions are changed. The measurements of dissociation constants (K_ds) between hydroxylase and mutated reductases confirmed that the binding affinities were not significantly changed, although the specific enzyme activities were significantly reduced by MMOR-Y93A. This result shows that Tyr93 could be a crucial residue for the electron transfer route at the interface between hydroxylase and reductase.

• Journal of Microbiology and Biotechnology
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• 제33권6호
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• pp.707-714
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• 2023

Plant-derived insecticide-neonicotinoid insecticides (NIs) played a crucial role in the development of agriculture and food industry in recent years. Nevertheless, synthesis of these nitrogen-containing heterocyclic compounds with an effective and greener routing remains challenging especially to the notion raise of "green chemistry" and "atom economy". While bio-catalyzed methods mediated by nicotinate dehydrogenase (NDHase) then provide an alternative. The current review mainly focuses on the introduction of sources, components, structure, catalytic mechanism and applications of NDHase. Specifically, NDHase is known as nicotinic acid hydroxylase and the sources principally derived from phylum Proteobacteria. In addition, NDHase requires the participation of the electron respiratory chain system on the cell membrane. And the most important components of the electron respiratory chain are hydrogen carrier, which is mainly composed of iron-sulfur proteins (Fe-S), flavin dehydrogenase (FAD), molybdenum binding protein and cytochromes. Heterologous expression studies were hampered by the plasmid and host with high efficiency and currently only Pseudomonas entomophila L48 as well as Comamonas testosterone was successfully utilized for the expression of NDHase. Furthermore, it is speculated that the conjugate and inductive effects of the substituent group at position 3 of the substrate pyridine ring exerts a critical role in the hydroxylation reactions at position 6 concerning about the substrate molecular recognition mechanism. Finally, applications of NDHase are addressed in terms of pesticide industry and wastewater treatment. On conclusion, this critical review would not only deepen our understanding of the theory about NDHase, but also provides the guideline for future investigation of NDHase.

• Journal of Nutrition and Health
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• 제55권3호
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• pp.321-329
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• 2022

리보플라빈 (비타민 B₂)은 한국인에게 섭취 부족의 우려가 있는 비타민으로서 에너지 대사를 포함한 산화/환원 반응의 조효소로서 작용한다. 적절한 섭취수준을 평가하는 방법으로는 적혈구에서의 리보플라빈 농도나 EGRAC, 혹은 소변의 리보플라빈 농도를 측정하는 것이다. 적혈구 리보플라빈 수준은 400 nmol/L (15 ㎍/100 mL) 이상을 적정수준, 270 nmol/L (10 ㎍/100 mL) 이하를 결핍으로 하거나 EGRAC 값이 1.2 이하인 경우 적정수준, 1.4이상이면 결핍으로 판정한다. 2020 한국인 영양소 섭취기준 개정 시 19-64세 성인의 2020 리보플라빈 평균필요량은 남자 1.3 mg/d, 여자 1.0 mg/d로 설정되었으며, 64-74세 남자 1.2 mg/d, 여자 0.9 mg/d, 75세 이상 남자 1.1 mg/d, 여자 0.8 mg/d로 성인 보다 낮게 설정되었다. 2020 국민건강영양조사 결과 19세 이상 우리나라 성인의 리보플라빈 평균 섭취량은 1.69 mg/d이며, 권장섭취량 대비 124.9% 였고, 보충제 섭취자들의 보충제로부터의 리보플라빈 섭취량은 평균 10.26 mg/d로 식품으로부터 섭취하는 1.50 mg에 비해 약 6.8배 높은 수준이었다 [18]. 2020 한국인 영양소 섭취기준에서 연령별, 성별 리보플라빈 권장섭취량은 각 생애주기 구간별 평균필요량에 변이계수 10%를 적용하여 평균필요량의 120% 수준으로 설정되었다 리보플라빈의 대표적 식품 급원으로는 유제품, 난류, 육류, 가금류와 생선류의 동물성 식품과 두류, 녹색채소류, 곡류 등이 있으며 2020 국민건강영양조사에 따르면 한국인의 리보플라빈 급원 식품으로 기여도가 높은 식품은 달걀, 라면, 돼지고기, 우유, 간장, 쇠고기, 배추김치, 닭고기, 고추가루, 시리얼 순이다. 최근 리보플라빈의 경우 비타민 보충제 외에도 다양한 건강기능성 식품섭취로 인해 보충제를 섭취하는 사람들의 경우 보충제로부터의 섭취수준이 식품으로부터의 섭취량을 훨씬 초과하고 있어 리보플라빈 영양상태 평가 시 보충제로부터 섭취하는 수준에 대한 평가가 향후 반드시 필요하다. 또한 2020 한국인 영양소 섭취기준 개정에서는 만성 질환 예방에 대한 부분은 아직 과학적 증거의 불충분으로 고려되지 않았으나 향후 만성질환과 관련된 역학연구 및 중재연구가 더 필요하다고 판단된다.

• 한국균학회지
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• 제17권3호
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• pp.161-168
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• 1989

표고버섯(L. edodes) 중의 mitochondria는 설탕밀도단계기울기법에 따라 분리정제 하였다. 앞서 보고한 바와 같이, 각 파장별 빛조사(400-700nm)에 따른 mitochondrial ATPase와 ATP synthase의 활성도는 680nm와 470nm에서 각각 활성화되었다. 본 연구에서, 400nm 이하의 파장별 빛조사에 따른 mitochondrial ATPase 및 ATP synthase의 활성도는 380nm와 330nm에서 각각 활성화되었으며, 330nm 및 350에서 각각 억제되었다. FAD의 존재하에서, mitochondrial ATP synthase는 활성화 파장 및 억제 파장의 조사에 의하여 활성도가 각각 증가된 반면, mitochondrial ATPase의 활성도는 감소되었다. 그러나, NADH의 존재하에서, 이들 파장의 조사에 의한 효소의 활성도는 변화가 없었다. 또한, 두 효소는 각각의 활성화 파장 및 억제 파장이 조사됨에 따라 $FADH_2$를 FAD로 산화시키는 spectrum을 보였다. 이로써, 이 두 효소는 빛 조사에 의하여 생체내의 산화 환원반응의 산화제로 작용하였으며, 특히 mitochondrial ATP synthase의 활성화에 따른 광유발물질은 mitochondria 중에 존재하는 flavin 또는 flavoprotein으로 추정된다.

• 한국수소및신에너지학회논문집
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• 제26권2호
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• pp.179-183
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• 2015

In this study, we propose a catalyst structure including enzyme and metal nano rod for glucose sensing. In the catalyst structure, glucose oxidase (GOx) and gold nano rod (GNR) are alternatingly immobilized on the surface of carbon nanotube (CNT), while poly(ethyleneimine) (PEI) is inserted in between the GOx and GNR to fortify their bonding and give them opposite polarization ($[GOx/GNR]_nPEI/CNT$). To investigate the impact of $[GOx/GNR]_nPEI/CNT$ on glucose sensing, some electrochemical measurements are carried out. Initially, their optimal layer is determined by using cyclic voltammogram and as a result of that, it is proved that $[GOx/GNR/PEI]_2/CNT$ is the best layer. Its glucose sensitivity is $13.315{\mu}AmM^{-1}cm^{-2}$. When it comes to the redox reaction mechanism of flavin adenine dinucleotide (FAD) within $[GOx/GNR/PEI]_2/CNT$, (i) oxygen plays a mediator role in moving electrons and protons generated by glucose oxidation reaction to those for the reduction reaction of FAD and (ii) glucose does not affect the redox reaction of FAD. It is also recognized that the $[GOx/GNR/PEI]_3/CNT$ is limited to the surface reaction and the reaction is quasi-reversible.

• 한국식품위생안전성학회지
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• 제37권4호
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• pp.207-215
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• 2022

계란의 비린내 강도는 가금류의 종류와 개인의 인식에 따라 다르지만 여러 요인에 의해 발생할 수 있다. 특히 트리메틸아민(TMA)이 원인으로 밝혀졌다. 주목할만한 점은 리보플라빈이 TMA를 무취의 트리메틸아민-N-옥사이드로 전환시키는 역할을 하는 효소인 플라빈 함유 모노옥시게나제 3의 활성을 증가시킬 수 있다는 점이다. 본 연구는 난황의 TMA 함량을 분석하여 비린내에 대한 기여도를 평가하고, 비린내를 방지하는 방법을 개발하는 것을 목적으로 하였다. 고체상 미세추출-기체 크로마토그래피/질량 분석법을 사용하여 리보플라빈이 강화, 보충된 사료를 먹인 암탉의 난황에 있는 휘발성 화합물을 감지하고 정량화했다. 또한 샘플 간 휘발성 물질의 상대적 함량을 비교하기 위해 전자코를 이용하여 상관관계 연구를 수행하였다. TMA는 콜린이 함유된사료를 섭취한 가금류의 난황에서 고농도로 검출되었지만 리보플라빈이 보충된 사료를 섭취한 가금류에서는 검출되지 않았다. 전반적으로, 이 연구는 리보플라빈이 TMA를 포함하여 계란에 존재하는 휘발성 물질의 양과 품질에 영향을 미친다는 것을 시사한다. 이러한 발견이 계란의 비린내를 줄이는 것은 물론 품질 향상에 기여하기를 기대한다.