• Proceedings of the Korean Society of Applied Entomolohy Conference
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• 2000.05a
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• pp.114-114
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• 2000

• Journal of Microbiology and Biotechnology
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• v.13 no.3
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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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• 2005.04a
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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.

• The Journal of Internal Korean Medicine
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• v.31 no.3
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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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• v.15 no.4
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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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• v.24
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• pp.29-35
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• 2006

In this study we evaluated the anti-diabetic potential of Puerariae Radix and its isoflavone (puerarin) by investigating their inhibitory activities against digestive enzymes, ${\alpha}$-amylase, ${\alpha}$-glucosidase and lipase and effect on glucose uptake and PPAR ${\gamma}$ expression. The activities of carbohydrate digestive enzymes were not inhibited by puerarin. Glucose uptake in differentiated adipocytes was stimulated by puerarin. Furthermore, puerarin enhanced the differentiation of preadipocytes as evaluated by triacylglycerol (TG) accumulation, which is specific for differentiated adipocytes. The effect of puerarin on expression of peroxisome proliferator-activated receptor ${\gamma}$ (PPAR-${\gamma}$) gene, which is associated with obesity and dyslipidemia, was examined by both real-time PCR and reverse transcriptase PCR. The study demonstrated that puerarin increased the expression of PPAR-${\gamma}$. In conclusion, puerarin showed potential to exert anti-diabetic action by enhancing cellular glucose uptake and thereby TG accumulation in adipocyte tissue.

• Journal of Life Science
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• v.23 no.11
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• pp.1404-1408
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• 2013

(-)-Epigallocatechin-3-gallate (EGCG), a green tea polyphenol, has been shown to have strong antibacterial, antiviral, antioxidant, anti-inflammatory, and chemopreventive effects. However it is unknown whether EGCG can recover alcohol-associated pancreatitis. The aim of this study was to investigate the effects of EGCG on pancreatic enzyme activities and the expressions of pancreatic regenerating related markers, such as adenosine monophosphate-activated protein kinase (AMPK), raf-1 kinase inhibitor protein (RKIP), and Regenerating gene 1 (Reg1), in mice pancreatic primary acinar cells. Our results revealed that activities of ${\alpha}$-amylase and chymotrypsin were significantly increased in the cells treated with ethanol compared to the untreated control cells; however, the increased activities of both enzymes were markedly reduced by pretreatment with EGCG. Phosphorylation of AMPK and total expression of RKIP were decreased in the ethanol-treated primary acinar cells; however, these were both significantly increased in the EGCG-pretreated cells. In addition, when EGCG was treated, expression of Reg1 was markedly increased compared with that of the control or the ethanol-treated primary acinar cells, demonstrating that EGCG can modulate pancreatic regenerating related genes. Therefore, our findings suggest that EGCG may have therapeutic utility in the prevention or treatment of alcohol-associated pancreatitis.

• Journal of Acupuncture Research
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• v.29 no.6
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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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• v.10 no.6
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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.

• The Korean Journal of Mycology
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• v.6 no.2
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• pp.1-10
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• 1978

In de novo biosynthesis of the extracellulor enzymes-proteinsaes, alpha and gluc-amylases during the synchronized differentiation of Aspergillus niger in submerged culture and surface liquid culture were investigated. Gluc-amylase was synthesized in the stage of presporulation in which phialide formation is involved. Proteinase was synthesized both in the stages of conidiophore formation and presporulation. Alpha-amylase was synthesized during presporulation and sporulation stages, the activity of enzyme lasted for seven days on surface liquid culture. It seemed that the synthesis was occured in de novo partly repressed by the catabolite, and its nature was found to be constitutive since it is produced in non-starch medium. Polyacrylamide gel electrophoresis have shown that presporulating and sporulating body produced diverse types of the proteins whereas the earlier stages of vegetative body showed simpler profiles. The uptake of C-14 uracil into RNA and C-14 glutamate into protein were shown to be vigorous in presporulating body rather than those in sporulating body. Coincidence of alpha-amylase biosynthesis in de novo and sporulation may be significant in the study of differentiation in which gene expression is involved.