• Title/Summary/Keyword: pET-Lip

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The Improvement of surface activity and Emulsification Activity by Transformation of Lipase Gene in Klebsiella sp. KCL-1, Oil-Degrading Bacterium. (Lipase gene의 도입에 의한 유류분해세균 Klebsiella sp. KCL-1의 표면활성과 유화력 향상)

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    • Journal of Life Science
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    • v.14 no.5
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    • pp.834-839
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    • 2004
  • To improve and oil degrading activity, the lipase gene from Pseudomonase sp. was transformed into Klebsiella sp. KCL-l, an oil degrading bacterium. The selected trasformant was named as a KCL-1/pET-Lip. The surface tension of culture broth of KCL-1/pET-Lip was decreased to 33 dyne/cm from 55 dyne/cm using 4% (v/v) soybean oil as sole carbon source. The surface tension were 44 and 37.5 dyne/cm, to 2% (w/v) glucose and 4% (v/v) kerosene medium, respectively. The emulsification activity of the biosurfactant solution containing lipase of KCL-l/pET-Lip improved better than wild type KCL-l. The soybean oil was most efficient carbon source and substrate for surface activity and emulsification activity of KCL-1/pET-Lip. The expression of lipase was confirmed by SDS-PAGE.

Expression and Purification of Three Lipases (LipAD1, LipAD2, and LipAD3) and a Lipase Chaperone (LipBD) from Acinetobacter schindleri DYL129 (Acinetobacter schindleri DYL129 유래의 3개 lipases와 chaperone의 발현과 정제)

  • Kim, Sun-Hee;Lee, Yong-Suk;Jeong, Hae-Rin;Pyeon, Hyo-Min;You, Ju-Soon;Choi, Yong-Lark
    • Journal of Life Science
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    • v.29 no.4
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    • pp.492-498
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    • 2019
  • Previously, three kinds of lipases, lipAD1, lipAD2, and lipAD3, and lipase chaperone, lipBD, of Acinetobacter schindleri DYL129 isolated from soil sample were reported. In this report, three lipase and lipase chaperone were cloned into the pET32a(+) or pGEX-6P-1 vectors for protein expression in Escherichia coli, and named as pETLAD1, pETLAD2, pETLAD3 and pETLBD or pGEXLAD1, pGEXLAD 2, pGEXLAD3 and pGEXLBD, respectively. Protein expression rate was higher in pET system than in pGEX system. Although LipAD1 and LipAD2 were produced as inclusion bodies, their expression levels were high. So LipAD1 and LipAD2 could be solubilized in 8 M urea buffer and purified. LipAD3 and LipBD were overexpressed in soluble form and purified. Those proteins were purified by His-tag affinity chromatography connected in AKTA prime system. The activities of the purified lipases were demonstrated with 1% tributyrin agar plate. After purification, molecular mass was determined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis. LipAD1 showed high activity toward ${\rho}$-nitrophenyl acetate and ${\rho}$-nitrophenyl butyrate, LipAD2 showed high activity toward ${\rho}$-nitrophenyl acetate and ${\rho}$-nitrophenyl myristate, and LipAD3 showed high activity toward ${\rho}$-nitrophenyl acetate, ${\rho}$-nitrophenyl butyrate, and ${\rho}$-nitrophenyl miristate, respectively. Three lipases, LipAD1, LipAD2, and LipAD3, showed optimal reaction at $50^{\circ}C$ using ${\rho}$-nitrophenyl butyrate, as substrate.

Isolation and Characterization of a Novel Triolein Selective Lipase from Soil Environmental Genes

  • Lim, Hee Kyung;Han, Ye-Jin;Hahm, Moon-Sun;Park, Soo Youl;Hwang, In Taek
    • Microbiology and Biotechnology Letters
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    • v.48 no.4
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    • pp.480-490
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    • 2020
  • A novel lipase gene, Lip-1420, was isolated from a metagenomic library constructed from reed marsh from Mt. Jumbong in Korea, comprising 112,500 members of recombinant plasmids. The DNA sequence of Lip-1420-subclone (5,513 bp) was found to contain at least 11 ORFs according to the GenBank database. The ORF-3 gene was inserted into the pET21a plasmid containing the C-terminal 6-His tag and transformed into E. coli BL21(DE3) to express the recombinant lipase protein. Lip-1420 was purified using a fast protein liquid chromatography system. The gene was registered in GenBank (MH628529). The values of Km and Vmax were determined as 0.268 mM and 1.821 units, respectively, at 40℃ and pH 8.0, using p-nitrophenyl palmitate as the substrate. This lipase belongs to family IV taxonomically because it has conserved HGGG and GDSAG motifs in the constitutive amino acid sequence. According to the predicted structural model, the binding sites are represented by residues H78, G81, D150, S151, A152, V181, and D236. Finally, Lip-1420 showed triolein selectivity for methanolysis between triolein (18:1) and tristearin (18:0) substrates. Further study of the selective mechanism and structure-function relationship of this new lipase could be useful for more practical applications.

Intra- and Interspecific Variation in the Ribosomal Internal Transcribed Spacers and 5.8S rDNA among three species of Pseudodiaptomus (Calanoida, Pseudodiaptomidae) from Korean Estuarine Waters

  • Soh, Ho-Young;Suh, Hae-Lip;Yoon, Yang-Ho;Park, Sang-Duk;Kwon, Soon-Yoo
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2001.05a
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    • pp.437-438
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    • 2001
  • Species of the genus Pseudodiaptomus are reported as the major constituents in the fresh or brackish waters in the northeastern Asia (Suh et al., 1991; Oka et al., 1991). Of these species, P. inopinus (Burckhardt) and P. poplesia (Shen) are particularly numerous in spring and autumn (Suh et al., 1991; Shen and Song, 1979). Recently. we found a presence of two types of Pseudodiaptomus inopinus during the course of investigation on zooplankton from Korean estuaries. (omitted)

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Case reports on neonatal mouth opening limitation due to congenital bilateral idiopathic hyperplasia of the coronoid processes (선천성 양측 특발성 오훼돌기 과형성으로 인한 개구장애의 증례보고)

  • Baek K.W.;Myung H.;Seo B.M.;Hwang S.J.;Lee J.H.;Choung P.H.;Kim M.J.;Choi J.Y.
    • Korean Journal of Cleft Lip And Palate
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    • v.7 no.2
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    • pp.133-139
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    • 2004
  • Congenital bilateral idiopathic hyperplasia of the coronoid processes presents with limited mouth-opening without visible maxillofacial deformity or temporomandibular joint dysfunction / disorder. According to Blanchard et al and McLoughlin et al, it was lnitially described in 1853 by Langenbeck, and the first cases were reported by Holmes in 1956. Since then, there have been regular reports of a certain number of cases. In 1995, McLoughlin et al recorded 79 published cases of bilateral hyperplasia of the coronoid processes. Among them, Fabie et al have found only 3 published cases relating to children younger than 8 years, and have presented the first case of objectively diagnosed restricted mouth opening from birth by pediatricians. Authors have experienced 2 child patients with mouth opening limitation who was diagnosed congenital bilateral idiopathic hyperplasia of the coronoid processes without any other TNJ morphology in Dept of Oral and Maxillofacial surgery, Seoul National University Dental Hospital in 2004. Coronoidectomy was performed and postoperative active mouth opening exercise is indicated. Authors report 2 cases of congenital bilateial idiopathic hypeiplasia of the coronoid processes with literature liview.

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Isolation, Cloning and Co-Expression of Lipase and Foldase Genes of Burkholderia territorii GP3 from Mount Papandayan Soil

  • Putra, Ludwinardo;Natadiputri, Griselda Herman;Meryandini, Anja;Suwanto, Antonius
    • Journal of Microbiology and Biotechnology
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    • v.29 no.6
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    • pp.944-951
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    • 2019
  • Lipases are industrial enzymes that catalyze both triglyceride hydrolysis and ester synthesis. The overexpression of lipase genes is considered one of the best approaches to increase the enzymatic production for industrial applications. Subfamily I.2. lipases require a chaperone or foldase in order to become a fully-activated enzyme. The goal of this research was to isolate, clone, and co-express genes that encode lipase and foldase from Burkholderia territorii GP3, a lipolytic bacterial isolate obtained from Mount Papandayan soil via growth on Soil Extract Rhodamine Agar. Genes that encode for lipase (lipBT) and foldase (lifBT) were successfully cloned from this isolate and co-expressed in the E. coli BL21 background. The highest expression was shown in E. coli BL21 (DE3) pLysS, using pET15b expression vector. LipBT was particulary unique as it showed highest activity with optimum temperature of $80^{\circ}C$ at pH 11.0. The optimum substrate for enzyme activity was $C_{10}$, which is highly stable in methanol solvent. The enzyme was strongly activated by $Ca^{2+}$, $Mg^{2+}$, and strongly inhibited by $Fe^{2+}$ and $Zn^{2+}$. In addition, the enzyme was stable and compatible in non-ionic surfactant, and was strongly incompatible in ionic surfactant.

Effects of dietary Gleditschia addition on biochemical composition of pork loin (조각자(주엽) 나무의 생리활성물질 Gleditschia이 비육돈육의 이화학적 성상에 미치는 영향)

  • Seo, Jong-Lip;Heo, Jung-Ho;Jung, Myung-Ho;Cho, Myung-Heui;Lee, Kuk-Cheon;Kim, Kuk-Hun;Hah, Dae-Shik;Ryu, Jae-Doo;Kim, Chung-Hui;Kim, Gon-Sup;Kim, Eui-Gyung;Kim, Jong-Shu
    • Korean Journal of Veterinary Research
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    • v.45 no.4
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    • pp.527-536
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    • 2005
  • The effects of dietary Gleditschia on fatty acid composition, lipid oxidation, and pork quality were investigated. Pigs (n=40) were fed a diet containing 0, 0.1, 0.2, 0.4, and 1% Gleditschia for 14 weeks and slaughtered at 110 kg average. The longissimus throracis et lumborum muscle was collected at 24 hr postmortem. Pork loin chops (3 cm thick) were packaged aerobically and stored at $4^{\circ}C$ for 7 days. Samples were analyzed for fatty acid composition, ultimate pH, thiobarbituric acid-reactive substance (TBARS), color ($L^*$, $a^*$, $b^*$), drip loss and water-holding capacity. There was no significant difference of moisture and crude fat percent in between treatment and control group, and the color stability of pork loin better in all treat groups was more improved during cold storage. The change of pH was significantly increased (p<0.05) in 0.2, 0.4, 1% treatment groups compare to the control group. Water-holding capacity of pig loins was significantly higher (p<0.05) in all treat groups than in control group; 0.2% treat group was the highest in the water holding capacity followed by 0.4, 1, and 0.1% treat groups. Less drip loss of pig loin was observed with samples from Gleditschia-fed pigs except 1% treat group. Unsaturated fatty acid were tend to be decreased and saturated fatty acid were tend to be increased in Gleditschia-treated group. Thiobarbituric acid-reactive substance value of control was significantly higher than that of the Gleditschia-fed group (p<0.05).

CLINICAL ANALYSIS OF GONIAL ANGLE CHANGE AFTER ORTHOGNATHIC SUGERY IN PATIENTS WITH THE MANDIBULAR PROGNATHISM (하악전돌증환자의 악교정수술후 하악각변화에 관한 임상적 분석)

  • Kwon, Yeong-Ho;Jang, Hyun-Jung;Lee, Sang-Han
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.22 no.2
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    • pp.206-216
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    • 2000
  • Predictional study for lateral change between pre- and post-orthognathic surgery has been emphasized mainly on anterior area of lateral profile; upper lip, lower lip and chin et al. So interest for lateral profile change has been less in posterior area of lateral profile and literature analyzing gonial angle change is rare. The purpose of this study is to make prediction for gonial angle change possible and to offer somewhat treatment guidance for gonial angle to be improved by investigating overall gonial angle change between pre- and post-orthognathic surgery and inquiring into factors influencing on pattern of genial angle change. For this study 35 patients were selected retrospectively. Lateral cephalometric radiographs were taken in just pre-op time, pod 1 day, pod 1 year. They were analyzed and genial angles were measured. The results were as follows : 1. Gonial angle at pod 1 day was decreased about $9.3^{\circ}$ than pre-op and gonial angle at pod 1 year was increased about $4.0^{\circ}$ than pod 1 day. So genial angle at pod 1 year was decreased about $5.3^{\circ}$ than pre-op genial angle(p<0.01). 2. Mean pre-op gonial angle was $129.4^{\circ}$, showing significantly high value than normal and mean gonial angle at pod 1 year was $124.1^{\circ}$, showing value near to normal. 3. Mean gonial angle change between pre-op and pod 1 year was decreased about $5.4^{\circ}$ in female and $5.3^{\circ}$ in male. There was no statistically significant difference between male and female(p>0.05). 4. Principal factor influencing on decreased gonial angle in gonial angle change between pre-op and pod 1 year was amount of mandibular setback. 5. Principal factor influencing on increased gonial angle in gonial angle change between pod 1 day and pod 1 year was % horizontal relapse, and it was thought that resorption and bone remodelling on posterior area in mandibular distal segment also were related to increased gonial angle. 6. It is thought that sagittal split ramus osteotomy in mandibular prognathic patients with high value of gonial angle is effective to improvement of gonial angle, and In patients who have normal range of gonial angle and are required with excessive mandibular setback, short lingual cut method, additional resection of posterior margin of distal segment, Obwegeser II method will be considerd. 7. More prudent operation and careful post-op management will be responsible for maintenance of postoperative stable gonial angle.

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