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Pichia pastoris에서 Zobellia galactanivorans 유래 재조합 $\beta$-Agarase의 고효율 분비생산 (High-level Secretory Expression of Recombinant $\beta$-Agarase from Zobellia galactanivorans in Pichia pastoris)

  • 석지환;박희균;이상현;남수완;전숭종;김종현;김연희
    • 한국미생물·생명공학회지
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    • 제38권1호
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    • pp.40-45
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    • 2010
  • Agarose의 $\beta$-1,4결함을 분해하는 Zobellia galactanivorans 유래의 $\beta$-agarase 유전자(agaB)는 클로닝 되었고, AOX1(alcohol oxidase 1, methanol inducible) promoter 하류에 Saccharomyces cerevisiae mating factor alpha-1 secretion signal($MF{\alpha}1$)를 연결하여 $MF{\alpha}1$-AgaB를 구축하였다. 구축된 plasmid pPIC-AgaB(9 kb)를 Pichia pastoris genome에 HIS4 gene 위치에 integration하였고, colony PCR을 통해 확인하였다. Methanol 첨가 배지에서 자란 형질전환체는 iodine solution의 첨가에 의해 red halos를 보였으며, P.pastoris에서 agaB의 효율적 분비 발현을 확인하였다. SDS-PAGE와 zymographic analysis에서 $\beta$-agarase의 분자량은 약 53 kDa으로 추정되었으며, 15% 정도의 N-linked glycosylation이 일어났음을 알 수 있었다. P.pastoris GS115/pPIC-AgaB의 48시간 baffled flask culture에서 세포외 $\beta$-agarase의 활성은 각각 0.1, 0.5, 1% methanol의 유도에 의해 1.34, 1.42 그리고 1.53 units/mL의 활성을 보였다. 대부분의 $\beta$-agarase의 활성은 세포 외에서 관찰되었고, 분비효율은 98%였으며 분비시의 glycosylation에 의해 열안정성도 증가되었다.

Role of LAMMER Kinase in Cell Wall Biogenesis during Vegetative Growth of Aspergillus nidulans

  • Choi, Yu Kyung;Kang, Eun-Hye;Park, Hee-Moon
    • Mycobiology
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    • 제42권4호
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    • pp.422-426
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    • 2014
  • Depending on the acquisition of developmental competence, the expression of genes for ${\beta}$-1,3-glucan synthase and chitin synthase was affected in different ways by Aspergillus nidulans LAMMER kinase. LAMMER kinase deletion, ${\Delta}lkhA$, led to decrease in ${\beta}$-1,3-glucan, but increase in chitin content. The ${\Delta}lkhA$ strain was also resistant to nikkomycin Z.

Bevacizumab accelerates corneal wound healing by inhibiting TGF-βexpression in alkali-burned mouse cornea

  • Lee, Sung-Ho;Leem, Hyun-Sung;Jeong, Seon-Mi;Lee, Koon-ja
    • BMB Reports
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    • 제42권12호
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    • pp.800-805
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    • 2009
  • This study investigated the effect of subconjunctival injections of bevacizumab, an anti-VEGF antibody, on processes involved in corneal wound healing after alkali burn injury. Mice were divided into three groups: Group 1 was the saline-treated control, group 2 received subconjunctival injection of bevacizumab 1hr after injury and group 3 received bevacizumab 1 hr and 4 days after injury. Cornea neovascularization and opacity were observed using a slit lamp microscope. Corneal repair was assessed through histological analysis and immunostaining for CD31, $\alpha$-SMA, collagen I, and TGF-$\beta$2 7 days post-injury. In group 3, injection of bevacizumab significantly lowered neovascularization and improved corneal transparency. Immunostaining analysis demonstrated a reduction in CD31, $\alpha$-SMA and TGF-$\beta$2 levels in stroma compared to group 1. These results indicate that bevacizumab may be useful in reducing neovascularization and improving corneal transparency following corneal alkali burn injury by accelerating regeneration of the basement membrane.

아시아티코사이드로부터 메칠 2${\beta},\;3{\beta}$-에폭시-23- 히드록시우르스-12-엔-28-오애이트의 합성 및 이의 환원적 에폭시환 개열반응 (Formation of Methyl 2${\beta},\;3{\beta}$-Epoxy-23-hydroxyurs-12-en-28-oate from Asiaticoside and Its Reductive Epoxide Ring Opening Reaction)

  • 주상섭;임두연;서성기;남태규;박형근;김희두;김창민;이민희;백형근;이민정;정영훈
    • 약학회지
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    • 제42권5호
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    • pp.500-506
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    • 1998
  • Steroidal 2${\beta},\;3{\beta}$--epoxy compound was prepared from asiaticoside via six steps and reduced regioselectively with lithium aluminum hydride. Epoxide ring opening furnished 9 as a sole product at reflux condition through axial hydride attack at C-3.

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저혈당을 경험한 당뇨병 환자의 치료지시 불이행, 가족지지가 우울에 미치는 영향 (Influence of Non-compliance of Treatment and Family Support on Depression in Diabetic Patients with Hypoglycemia)

  • 손혜경;정경숙
    • 한국콘텐츠학회논문지
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    • 제18권6호
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    • pp.518-528
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    • 2018
  • 본 연구는 저혈당을 경험한 당뇨병 환자의 우울에 미치는 영향요인을 파악하여 대상자의 우울을 감소시키기 위한 기초자료를 마련하고자 2015년 10월 1일부터 2016년 3월 15일까지 B광역시 소재 일 병원의 내분비내과 외래를 방문한 119명을 대상으로 설문지를 배부하여 자료를 수집하였다. 수집된 자료는 SPSS 20.0 프로그램을 이용하여 빈도와 백분율, t-검증, 일원분산분석, 사후검증(Scheffe's-test), Pearson's correlation, 단계적 다중회귀분석을 실시하였다. 본 연구 결과 대상자의 우울은 치료지시불이행(r=.42, p<.001)과는 양의 상관관계를, 가족지지(r=-.38, p<.001)와는 음의 상관관계를 보였다. 대상자의 우울에 영향을 미치는 요인은 가족지지(${\beta}=-.29$, p<.001), 치료지시불이행의 하위영역인 약물요법(${\beta}=.21$, p=.020), 식이요법(${\beta}=.23$, p=.014), 한 달 평균 수입 300만원 이상(${\beta}=-.20$, p=.014), 한 달 평균 수입 200만원 이상-300만원 미만(${\beta}=-.17$, p=.033)으로 확인되었으며, 총 30.0%를 설명하였다(Adj. R2=.30, F=20.06, p<.001). 이를 바탕으로 대상자의 우울을 감소시키기 위하여 가족지지 향상 프로그램을 개발하여 적용할 것을 제안한다.

단풍취로부터 분리한 Apigenin $7-O-{\beta}-D-glucoside$가 알콜대사효소에 미치는 영향 (Effects of Compounds Isolated from Ainsliaea acerifolia on the Hepatic Alcohol Dehydrogenase Activity)

  • 지옥표;신말식;문형인
    • Applied Biological Chemistry
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    • 제42권2호
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    • pp.162-165
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    • 1999
  • Spraque-Dawley계 수컷랫트에 단풍취로부터 분리한 화합물을 경구투여하고 혈청 ethanol농도와 간의 ADH 활성에 미치는 효과를 검토한 결과 알코올대사를 촉진시키는 성분은 apigenin $7-O-{\beta}-D-glucoside$로 확인되었다.

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Blockade of Trigeminal Glutamate Recycling Produces Anti-allodynic Effects in Rats with Inflammatory and Neuropathic Pain

  • Yang, Kui-Ye;Lee, Min-Kyung;Park, Min-Kyoung;Son, Jo-Young;Ju, Jin-Sook;Ahn, Dong-Kuk
    • International Journal of Oral Biology
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    • 제42권3호
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    • pp.129-135
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    • 2017
  • The present study investigated the role of spinal glutamate recycling in the development of orofacial inflammatory pain or trigeminal neuropathic pain. Experiments were carried out on male Sprague-Dawley rats weighing between 230 and 280 g. Under anesthesia, a polyethylene tube was implanted in the atlanto-occipital membrane for intracisternal administration. IL-$1{\beta}$-induced inflammation was employed as an orofacial acute inflammatory pain model. IL-$1{\beta}$ (10 ng) was injected subcutaneously into one vibrissal pad. We used the trigeminal neuropathic pain animal model produced by chronic constriction injury of the infraorbital nerve. DL-threo-${\beta}$-benzyloxyaspartate (TBOA) or methionine sulfoximine (MSO) was administered intracisternally to block the spinal glutamate transporter and the glutamine synthetase activity in astroglia. Intracisternal administration of TBOA produced mechanical allodynia in naïve rats, but it significantly attenuated mechanical allodynia in rats with interleukin (IL)-$1{\beta}$-induced inflammatory pain or trigeminal neuropathic pain. In contrast, intracisternal injection of MSO produced anti-allodynic effects in rats treated with IL-$1{\beta}$ or with infraorbital nerve injury. Intracisternal administration of MSO did not produce mechanical allodynia in naive rats. These results suggest that blockade of glutamate recycling induced pro-nociception in na?ve rats, but it paradoxically resulted in anti-nociception in rats experiencing inflammatory or neuropathic pain. Moreover, blockade of glutamate reuptake could represent a new therapeutic target for the treatment of chronic pain conditions.

Sleep Quality and Attention May Correlate With Hand Grip Strength: FARM Study

  • Lee, Gyuhyun;Baek, Sora;Park, Hee-won;Kang, Eun Kyoung
    • Annals of Rehabilitation Medicine
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    • 제42권6호
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    • pp.822-832
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    • 2018
  • Objective To determine the socio-demographic, psychologic, hematologic, or other relevant factors associated with hand grip strength in Korean farmers. Methods A total of 528 healthy Korean farmers were enrolled. Hand grip strength was measured in both hands using a hydraulic dynamometer. Socio-demographic characteristics were assessed and anthropometric measurements were obtained. Psycho-cognitive measurements such as sleep quality (Pittsburgh Sleep Quality Index) and Go/No-Go test response time were conducted. In addition to physical measurements, serologic parameters including insulin-like growth factor 1 were measured. The factors associated with hand grip strength were analyzed using multiple linear regression analysis after adjusting for age, height, and weight. Results The mean hand grip strength was associated with the Pittsburgh Sleep Quality Index total score (${\beta}=-0.12$, p=0.01), the Go/No-Go test response time (${\beta}=-0.18$, p=0.001), vitamin D (${\beta}=0.12$, p=0.02), and insulin-like growth factor 1 levels (${\beta}=0.1$, p=0.045). In female farmers, hand grip strength was only associated with the Pittsburgh Sleep Quality Index total score (${\beta}=-0.32$, p<0.001). Conclusion The results of this study demonstrate that hand grip strength was associated with sleep quality and attention in Korean farmers.

Pullulanase의 Reverse Reaction을 이용한 Maltosyl-$\beta$-Cyclodextrin의 합성 (Synthesis of Maltosyl-$\beta$-Cyclodextrin through the Reverse Reaction of Pullulanase)

  • 한일근;이용현
    • 한국미생물·생명공학회지
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    • 제19권5호
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    • pp.444-449
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    • 1991
  • Pullulanase의 역반응능을 이용하여 maltose와 $\beta$-cyclodextrin으로부터 maltosyl-$\beta$-cyclodextrin을 중합합성하기 위한 최적효소반응조건을 검토하였다. Maltose와 $\beta$-CD를 기질로 maltosyl-$\beta$-CD을 합성하였을 경우, 기질의 농도 70( w/w, 70g/100ml $H2_O$ ), malto-loigo당 /$\beta$-CD의 혼합비 12.7, 그리고 사용효소량 350 units/100ml일 때 최대전융인 43(w/w, g branched-CD/g CD)를 얻었고, 생성량은 2.31g/100ml였다. Maltosyl-$\beta$-CD의 효소합성의 적정 pH 및 온도는 각각 4.9와 $60^{\circ}C$ 였다. 또한 maltose와 $\alpha ,\beta$-그리고 $\gamma$-CD 각각을 기질로하여 maltosyl $\Alpha, \beta$ 그리고 $\gamma$-CD를 합성하였을 경우 전환율은 51.8, 42.6, 그리고 48.1로써, 생성량은 각각 2.8, 2.3 그리고 2.6g/100ml였다.

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A Novel Esterase from Paenibacillus sp. PBS-2 Is a New Member of the ${\beta}$-Lactamase Belonging to the Family VIII Lipases/Esterases

  • Kim, Young-Ok;Park, In-Suk;Nam, Bo-Hye;Kim, Dong-Gyun;Jee, Young-Ju;Lee, Sang-Jun;An, Cheul-Min
    • Journal of Microbiology and Biotechnology
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    • 제24권9호
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    • pp.1260-1268
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    • 2014
  • Screening of a gene library from Paenibacillus sp. PBS-2 generated in Escherichia coli led to the identification of a clone with lipolytic activity. Sequence analysis showed an open reading frame encoding a polypeptide of 378 amino acid residues with a predicted molecular mass of 42 kDa. The esterase displayed 69% and 42% identity with the putative ${\beta}$-lactamases from Paenibacillus sp. JDR-2 and Clostridium sp. BNL1100, respectively. The esterase contained a Ser-x-x-Lys motif that is conserved among all ${\beta}$-lactamases found to date. The protein PBS-2 was produced in both soluble and insoluble forms when E. coli cells harboring the gene were cultured at $18^{\circ}C$. The enzyme is a serine protein and was active against p-nitrophenyl esters of $C_2$, $C_4$, $C_8$, and $C_{10}$. The optimum pH and temperature for enzyme activity were pH 9.0 and $30^{\circ}C$, respectively. Relative activity of 55% remained at up to $5^{\circ}C$ with an activation energy of 5.84 kcal/mol, which indicates that the enzyme is cold-adapted. Enzyme activity was inhibited by $Cd^{2+}$, $Cu^{2+}$, and $Hg^{2+}$ ions. As expected for a serine esterase, activity was inhibited by phenylmethylsulfonyl fluoride. The enzyme was remarkably active and stable in the presence of commercial detergents and organic solvents. This cold-adapted esterase has potential as a biocatalyst and detergent additive for use at low temperatures.