• 한국응용곤충학회:학술대회논문집
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• 한국응용곤충학회 2000년도 합동 춘계 학술발표회
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• pp.114-114
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• 2000

• Journal of Microbiology and Biotechnology
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• 제13권3호
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• pp.451-456
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• 2003

An expression vector system was developed for the secretory production of recombinant Bacillus stearothermophilus L1 lipase in Saccharomyces cerevisiae. The mature L1 lipase gene was fused to ${\alpha}-amylase$ signal sequence from Aspergillus oryzae for the effective secretion into the culture broth and the expression was controlled under GAL10 (the gene coding UDP-galactose epimerase of S. cerevisiae) promoter. S. cerevisiae harboring the resulting plasmid successfully secreted L1 lipase into the culture broth. To examine an optimum condition for L1 lipase expression in the fed-batch culture, L1 lipase expression was induced at three different growth phases (early, mid, and late-exponential growth phases). Maximum product on of L1 lipase (1,254,000 U/l, corresponding to 0.65/1) was found when the culture was induced at an early growth phase. Secreted recombinant L1 lipase was purified only through CM-Sepharose chromatography, and the purified enzyme showed 1,963 U/mg of specific activity and thermoalkalophilic properties similar to those reported for the enzyme expressed in Escherichia coli.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2005년도 생물공학의 동향(XVI)
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• pp.402-406
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• 2005

Lipomyces starkeyi produces a novel glucanhydrolase containing endo-dextranase and ${\alpha}-amylase$ activities. A cDNA from L. starkeyi encoding a dextranase was isolated and characterized. The 2,052 kb cDNA fragment (lsd1) carrying dextranase gene showed one open reading frame (ORF) composed of 1,824 bp flanked by a 41 bp 5'-UTR and a 184 bp 3'-UTR including a poly(A) tail of 27 bp. The ORF encodes for a 608 amino acid with a predicted molecular mass of 67.6 kDa. There was 77% deduced amino acid sequence identity between the LSD1 dextranase and the dextranase from Penicillium minioluteum. The primary structure of the dextranase from L. starkeyi has distant similarity with enzymes belonging to glycosyl hydrolase family 49. The lsd1 protein was expressed in the Saccharmyces cerevisiae under control of GAL1 promoter and active dextranase was produced.

• 대한한방내과학회지
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• 제31권3호
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• pp.500-512
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• 2010

Objectives : This study was designed to investigate the effects of SuJeom-san(SJS) extract in rats with caerulein-induced acute pancreatitis (AP). Methods : We examined changes of pancreatic weight, histological, immunohistochemical and gene expression of cyclooxygenase (COX-2). Thirty-six adult male Sprague-Dawley rats were divided into six groups as follow: normal(Nor), caerulein-induced (Con), caerulein + cefotaxime sodium(CT), caerulein + SJS 3 mg/kg(SJSA), caerulein + SJS 6 mg/kg(SJSB) and caerulein + SJS 12 mg/kg(SJSC) groups. Pancreatic tissues of rats from all groups were removed for histological observation and light, and electron microscopic examination. Platelet activating factor(PAF) and Interleukin-6(IL-6) levels were determined spectrophotometrically. Results : The ratio of pancreas/body weight was significantly(p<0.05) increased in the Con compared with Nor, but significantly(p<0.05) decreased in SJSA, SJSB, SJSC and CT groups compared with Con. Caerulein administration significantly increased(p<0.05) the levels of amylase, but SJSA, SJSB, SJSC and CT significantly(p<0.05) reduced the levels of these enzymes. The levels of platelet activating factor(PAF) increased in Con compared with Nor, but decreased in SJSA, SJSB, SJSC and CT groups compared with Con. Interleukin-6(IL-6) levels increased significantly in all groups compared to Nor at 6 hrs, but significantly(p<0.05) reduced in SJSA, SJSB, SJSC and CT groups compared with Con at 24 hrs. The levels of tumor necrosis factor(TNF)-${\alpha}$ levels increased in all groups compared to Nor at 6 hrs, but significantly(p<0.05) reduced in SJSA, SJSB, SJSC and CT groups compared with Con at 24 hrs. The COX-2 positive materials were observed in the pancreas of the Con, but these positive materials were decreased in the SJS extract treatment group. Conclusion : SJS is potentially capable of limiting pancreatic damage during AP by restoring the fine structure of acinar cells and tissue; therefore, we conclude that SJS may have beneficial effects in the treatment of caerulein-induced AP.

• Preventive Nutrition and Food Science
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• 제15권4호
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• pp.360-363
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• 2010

To enhance the expression and secretion of pediocin PA-1 from heterologous bacterial hosts, the promoter and deduced signal sequence (PS) of an $\alpha$-amylase gene from a Bifidobacterium adolescentis strain was fused with pediocin PA-1 structural and immunity genes (AB) and the resulting functions were evaluated in Escherichia coli. Two recombinant PCR products were created-one with just the deduced signal sequence and one with the sequence plus the Ser and Thr sequences that are the next two amino acids of the signal sequence. These two products, the PSAB (---AQA::KYY---) and PSABST (---AQA$\underline{ST}$::KYY---), respectively, were inserted into a TA cloning vector (yT&A) and named pPSAB, which was previously reported, and pPSABST. The two recombinant plasmid DNAs were transferred into E. coli JM109 and the transformants displayed antimicrobial activity, where the activity of E. coli JM109 (pPSAB) was stronger than that of E. coli JM109 (pPSABST), indicating that the ST amino acid residues were not necessary for secretion and might have even decreased the antimicrobial activity of recombinant pediocin PA-1.

• Current Research on Agriculture and Life Sciences
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• 제24권
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• pp.29-35
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• 2006

본 연구에서는 갈근 및 그의 주 이소플라본인 puerarin의 활성을 세포 수준에서 분석하였다. 먼저 갈근에 함유된 이소플라본의 양을 분석한 결과, puerarin이 총 이소플라본의 90 % 차지하였다. 다음으로는 puerarin의 항당뇨 활성을 검정한 결과 먼저 탄수화물 및 지방소화효소 저해활성에 대해서는 거의 미비한 것으로 나타났으나 인슐린 감수성 및 지방세포의 분화의 유도에 대해서는 농도 의존적으로 작용하는 것으로 관찰되었다. 따라서 puerarin은 지방조직내로 포도당의 흡수를 촉진함으로서 항당뇨 효능을 발휘하는 것으로 추정된다.

• 생명과학회지
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• 제23권11호
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• pp.1404-1408
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• 2013

본 연구에서는 마우스 췌장의 선포세포(primary acinar cell)에서 에탄올에 의해 유도된 염증성 손상에 미치는 녹차의 생리활성 물질인 EGCG의 효과를 췌장분비 효소의 활성과 선포세포에서의 기능 회복 관련 마크로 알려진 유전자들(AMPK, RKIP 및 Reg1)의 발현 조절 측면에서 조사하였다. 대조군에 비해 에탄올이 처리된 세포에서는 ${\alpha}$-amylase와 chymotrypsin의 활성이 증가되었지만, EGCG 처리에 의하여 그들 활성이 유의적으로 감소하였다. 세포 생존 및 보호와 관련 있는 AMPK 인산화 수준은 에탄올 단독 처리시 그 발현이 감소하였지만, EGCG 처리에 의하여 유의적으로 회복되었다. 세포 사멸과 세포독성 조절과 연관이 있는 RKIP 또한 그 발현 경향이 AMPK인산화 변화의 정도와 유사하였다. 또한 베타세포 재생에 관여하는 Reg1 발현은 에탄올이 처리된 선포세포에 EGCG를 처리하였을 경우 그 발현량이 유의적으로 증가하였다. 이상의 결과들을 종합하여, 항산화성 폴리페놀인 EGCG는 알코올성 췌장염으로 인한 세포의 손상이나 당뇨병 예방과 회복에 치료적 효과를 가질 수 있다고 제안한다.

• Journal of Acupuncture Research
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• 제29권6호
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• pp.35-45
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• 2012

Objectives : This study was designed to investigate Effects of Sujeom powder(SJP) pharmacopuncture Injected at Jung-wan($CV_{12}$) in rats with caerulein-induced acute pancreatitis(AP). Methods : We examined changes of organ weight, histology, immunohistochemistry and gene expression of cycolooxygenase 2(COX-2) in the pancreas. Twenty adult male Sprague-Dawley rats were divided into four groups as follow: normal(Nor), caerulein-induced(Con), caerulein+SJP pharmacopuncture 0.2mL injected at Jung-wan($CV_{12}$)(SA), and caerulein+SJP pharmacopuncture 0.8 mL injected at Jung-wan($CV_{12}$)(SB) groups. Pancreatic tissues of rats from all groups were removed for histological observation and light microscopic examination. Interleukin-6(IL-6) levels were determined spectrophotometrically. Results : The ratio of pancreas/body weights was significantly(p<0.05) increased in the Con, the SA and the SB compared with the Nor, but was slightly decreased in the SA and in the SB groups compared with the Con. Caerulein administration has significantly(p<0.05) increased in the levels of amylase, but the SA, the SB significantly(p<0.05) decreased in the levels of these enzyme. The levels of amylase were increased significantly with caerulein administration, but were inhibited significantly in the SA and in the SB groups. Interleukin-6(IL-6) levels were significantly(p<005) increased in all groups compared with the Nor, especially in the SB. were significantly increased. The levels of Tumor necrosis factor(TNF)-${\alpha}$ levels were significantly increased in all groups compared with the Nor. In the conclusion, the datum of IL-6 and TNF-${\alpha}$ are suggested that the inflamation was still existed actively at a point of measurement(24 hours later). The COX-2 positive materials are observed in the pancreas from the Con, but these positive materials are decreased in the SJP pharmacopuncture at Jung-wan($CV_{12}$) treatment group. Conclusion : SJP pharmacopuncture injected at Jung-wan($CV_{12}$) is potentially capable of limiting pancreatic damage during AP by restoring the fine structure of acinar cells and tissues. Therefore we can say that SJP pharmacopuncture Injected at Jung-wan($CV_{12}$) may have beneficial effects in the treatment of caerulein-induded AP. Further studies about the adequate amount of the SJP pharmacopuncture and about more effective route of administration is still required.

• Biotechnology and Bioprocess Engineering:BBE
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• 제10권6호
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• pp.576-581
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• 2005

Phytase improves the bioavailability of phytate phosphorus in plant foods to humans and animals, and reduces the phosphorus pollution of animal waste. We have engineered the cell surface of the yeast. Saccharomyces cerevisiae, by anchoring active fungal phytase on its cell wall, in order to apply it as a dietary supplement containing bioconversional functions in animal foods and a whole cell bio-catalyst for the treatment of waste. The phytase gene (phyA) of Aspergillus niger with a signal peptide of rice amylase 1A (Ramy1A) was fused with the gene encoding the C-terminal half (320 amino acid residues from the C-terminus) of yeast ${\alpha}-agglutinin$, a protein which is involved in mating and is covalently anchored to the cell wall. The resulting fusion construct was introduced into S. cerevisiae and expressed under the control of the constitutive glyceraldehydes-3-phosphate dehydrogenase (GPD) promoter. Phytase plate assay revealed that the surface-engineered cell exhibited a catalytically active opaque zone which was restricted to the margin of the colony. Additionally, the phytase activity was detected in the cell fraction, but was not detected in the culture medium when it was grown in liquid. These results indicate that the phytase was successfully anchored to the cell surface of yeast and was displayed as its active form. The amount of recombinant phytase on the surface of yeast cells was estimated to be 16,000 molecules per cell.

• 한국균학회지
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• 제6권2호
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• pp.1-10
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• 1978

검정곰팡이(Aspergillus niger)의 액침배양과 액체표면배양을 통한 동조적 형태분화에 있어서 체외효소인 단백질분해효소, 알파 및 굴루크아밀라제의 신생적 생합성 상황을 연구하였다. 굴루크아미라제는 경자(phialide)가 성숙하는 단계 즉 포자 형성의 전단계에서만 그 활성이 왕성하였다. 단백질분해효소(산성, 중성 및 알칼리성)들은 분생자병의 성장단계에서 활성이 약간 증가하였으나, 경자의 성숙단계에서는 활성이 극히 활발하였다. 알파아밀라제는 경자의 성숙시기와 포자형성기에서 활성이 활발하였으며 그 활성은 장기간 지속 되었다. 알파아밀라제의 활성은 포자형성 기간중 계속 증가하였으므로 신생적으로 생합성된다고 할 수 있으며, 포도당배지에서 많은 량이 생합성되었고, 또 포도당량의 고갈에 즈음하여 그 생합성이 개시되었으므로 이 효소는 구성적 효소이며 이화 대사물의 억제작용(catabolite repression)을 받는 효소라고 할 수 있다. 포리아크릴아미드 젤(polyacrylamide gel)을 이용한 전기영동으로서 포자형성기와 그 전단계의 균체로부터 다양하고 선명한 세포외성 단백질을 분리할 수 있었다. 균사형성기나 포자발아기의 균체로부터는 극소수의 선명치 못한 분리상을 얻었다. C-14 우라실이 균체의 RNA핵산으로 섭취되어 들어가는 비율과 C-14 굴루탐산이 균체 단백질으로 섭취되어 들어가는 비율은 포자형성전기에서 왕성하였으며 포자형성 기간중에는 극히 저조하였다. 알파아밀라제의 신생적 생합성과 포자형성이 일치하는 현상은 유전인자의 표현과정이 내포되는 분화(포자형성)라는 점에서 볼 때 의의와 인과관계가 있을 것으로 사료된다.(1.9 eV)와 $e_g$ $(2.8{\sim}3.0\;eV)$로의 전이 즉, $O^{2-}(2p){\rightarrow}Mn^{4+}(3d)$ 및 $O^{2-}$에서 $Mn^{3+}$ 이온의 $t_{2g}$ (2.3 eV)와 $e_g$ ($3.4{\sim}3.6$ eV)로의 전이 즉, $O^{2-}(2p){\rightarrow}Mn^{3+}(3d)$ 등에 의한 것으로 해석된다. 또한, 1.6, 1.8, 1.9 eV 부근에서 관측된 좁은 에너지 영역의 흡수구조 들은 팔면체 $Mn^{3+}$ 이온 내에서의 d-d 결정장(crystal-field) 전이에 의한 것으로 해석된다. 이러한 흡수구조는 Ni 치환량이 증가함에 따라 그 강도가 감소한다. x = 0.6의 경우 $e_g$ 상태와 관련된 CT 전이구조 들이 $t_{2g}$ 상태와 관련된 전이구조 들에 비하여 큰 폭으로 감소하는데 이것은 Jahn-Teller 효과에 의해서 격자상수가 tetragonal 구조로 확장됨에 따라 $e_g$ 상태와 $O^{2-}(2p)$ 상태 간의 파동함수 중첩이 감소한 것에 기인하는 것으로 해석된다.)의 영향(影響)을 더 많이 받고 있었다. 마. total ginsenosides의 분해반응시(分解反應時)의 활성화(活性化)에너지($E_a$)는 17.7kcal/mole이었고