• Title/Summary/Keyword: inhibition layer

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EFFECT OF CHLORHEXIDINE ON MICROTENSILE BOND STRENGTH OF DENTIN BONDING SYSTEMS (Chlorhexidine 처리가 상아질 접착제의 미세인장결합강도에 미치는 영향)

  • Oh, Eun-Hwa;Choi, Kyoung-Kyu;Kim, Jong-Ryul;Park, Sang-Jin
    • Restorative Dentistry and Endodontics
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    • v.33 no.2
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    • pp.148-161
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    • 2008
  • The purpose of this study was to evaluate the effect of chlorhexidine (CHX) on microtensile bond strength (${\mu}TBS$) of dentin bonding systems. Dentin collagenolytic and gelatinolytic activities can be suppressed by protease inhibitors, indicating that MMPs (Matrix metalloproteinases) inhibition could be beneficial in the preservation of hybrid layers. Chlorhexidine (CHX) is known as an inhibitor of MMPs activity in vitro. The experiment was proceeded as follows: At first, flat occlusal surfaces were prepared on mid-coronal dentin of extracted third molars. GI (Glass Ionomer) group was treated with dentin conditioner, and then, applied with 2 % CHX. Both SM (Scotchbond Multipurpose) and SB (Single Bond) group were applied with CHX after acid-etched with 37% phosphoric acid. TS (Clearfil Tri-S) group was applied with CHX, and then, with adhesives. Hybrid composite Z-250 and resin-modified glass ionomer Fuji-II LC was built up on experimental dentin surfaces. Half of them were subjected to 10,000 thermocycle, while the others were tested immediately. With the resulting data, statistically two-way ANOVA was performed to assess the ${\mu}TBS$ before and after thermo cycling and the effect of CHX. All statistical tests were carried out at the 95 % level of confidence. The failure mode of the testing samples was observed under a scanning electron microscopy (SEM). Within limited results, the results of this study were as follows; 1. In all experimental groups applied with 2 % chlorhexidine, the microtensile bond strength increased, and thermo cycling decreased the micro tensile bond strength (P > 0.05). 2. Compared to the thermocycling groups without chlorhexidine, those with both thermocycling and chlorhexidine showed higher microtensile bond strength, and there was significant difference especially in GI and TS groups. 3. SEM analysis of failure mode distribution revealed the adhesive failure at hybrid layer in most of the specimen. and the shift of the failure site from bottom to top of the hybrid layer with chlorhexidine groups. 2 % chlorhexidine application after acid-etching proved to preserve the durability of the hybrid layer and microtensile bond strength of dentin bonding systems.

A study on screeining of antibacterial herb medicines against puerperal infection-bacteria, Proteus vulgaris, Staphylococcus aures, and Enterococcus faecalis (산욕기(産褥期) 감염(感染) 유발(誘發) 세균(細菌)의 생육(生育)을 억제하는 한약재(漢藥材) 탐색(探索)에 관(關)한 연구(硏究))

  • Lim, Je-Yeon;Seong, Yeon-Su;Kim, Hee-Jin;Lee, Tae-Gyun
    • The Journal of Dong Guk Oriental Medicine
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    • v.6 no.1
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    • pp.177-203
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    • 1997
  • Various kinds of medicinal herbs and prescribed herb medicines which have usually been used for treatment of reducing fever, purging intense heat, and detoxication were secreened for antibacterial activity against Proteus vulgaris, S. aureus, and E. faecalis which causing puerperal infection. Minimal inhibitory concentrations of water and ethanol-soluble extracts of Hwangyon(C. japonica), Omae(P. mume), and Omiza(S. chinensis) were determined and using thin-layer chromatography the extracts of Hwangyon and Omiza were separated. The results obtained were as follow : 1. The water-soluble extracts of Keumeunhwa(L. japonica), Chiza(G. jasminoides), Hwangyon(C. japonica), Dansam(S. miltiorrhiza), Omae(P. mume) and Omiza(S. chinensis) showed antibacterial activities against P. vulgaris ; among them, the extracts of Hwangyon, Omae, and Omiza showed high antibacterial activities. The water-soluble extracts of Keumeunhwa(L. japonica), Hwangyon(C. japonica), Hwangkeum(S. baicalensis), Hwangbaek(P. amurense), Dansam(S. miltiorrhiza), Daehwang(R. coreanum), Omae(P. mume), Omiza(S. chinensis), Gingseng(P. ginseng), and Gamcho(G. uralensis) showed antibacterial activities against S. aureus : among them, the extracts of Hwangyon and Omiza showed high antibacterial activities. The water-soluble extracts of Banggi(C. trilobus), Daehwang(R. coreanum), Omae(P. mume), Omiza(S. chinensis), and Gamcho(G. uralensis) inhibited the cell growth of E. faecalis : among them, Omae and Omiza showed high antibacterial activities. The extract of Hwangyon(C. Japonica) and Omae(P. mume) showed no inhibition against E. coli. However, the extract of Omiza(S. chinensis) inhibited cell growth of both E. coli and B. subtilis. 2. The water- and ethanol-soluble extract of Hwangyon(C. Japonica) showed antibacterial activities against P. vulgaris and S. aureus, those of Omae(P. mume) against P. vulgaris and E. faecalis, and those of Omiza(S. chinensis) against all species tested, P. vulgaris, S. aureus, and E. faecalis. With the exception that ethanol-soluble extract of Hwangyon(C. japonica) showed much higher antibacterial activity against S. aureus than water-soluble one, antibacterial activities of both water-and ethanol-soluble ones were similar to each other of other two medicinal herbs. 3. When the prescribed herb medicines were tested, Sambohwan showed antibacterial activities against P. vulgaris and S. aureus. No prescribed herb medicine inhibitied the cell growth of E. faecalis. 4. Minimal inhibitory concentrations(MICs) of both water- and ethanol-soluble extracts of Hwangyon(C. japonica), Omae(P. mume), and Omiza(S. chinensis) against P. vulgaris were $2.5mg/m{\ell}$, $10mm{\ell}$, and $20mg/m{\ell}$, respectively. Those of both Hwangyon(C. japonica) and Omiza(S. chinensis) against S. aureus were $1.25mg/m{\ell}$ and $10mg/m{\ell}$, respectively. MICs of water-soluble extracts of Omae(P. mume) and Omiza(S. chinensis) against E. faecalis were $2.5m{\ell}$ and $5m{\ell}$, and those of ethanol-soluble extracts were $5m{\ell}$ and $10m{\ell}$, respectively Except for those of E. faecalis the cell growth of P. vulgaris and S. aureus were inhibited by much lower concentration of ethanol-soluble extracts used. As a result, the antibacterial compounds against P. vulgaris S. aureus, and E. aecalis are contained in the extracts of Hwangyon(C. japonica), Omae(P. mume), Omiza(S. chinensis), the prescribed herb medicine, Sambohwan, and might be used for treatment of puerperal infection. Further study should be carried out to identify which compounds affect the cell growth inhibition of P. vulgaris S. aureus, and E. faecalis.

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Inhibition of Cancer Cell Migration by Compounds from Garlic Extracts (마늘추출물에 의한 암세포의 이동 저하)

  • Kim, Eun-Kyoung;Yun, Sung-Ji;Ha, Jung-Min;Jin, In-Hye;Kim, Young-Whan;Kim, Sun-Gun;Park, Da-Jung;Choi, Young-Whan;Yun, Sik;Kim, Chi-Dae;Bae, Sun-Sik
    • Journal of Life Science
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    • v.21 no.6
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    • pp.767-774
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    • 2011
  • Cell migration plays a fundamental role in cancer cell invasion and metastasis as well as in many physiological responses. Here, we screened four different sources of garlic - water extract of normal and black garlic, as well as dried normal and black garlic - for the identification of anti-invasive and anti-metastatic activity on cancer cells. Inhibition of cancer cell migration was observed in the hexane extract of dried-garlic. Inhibitory activity was further purified to near homogeneity by thin layer chromatography and named $\b{i}$nhibitor of $\b{c}$ancer $\b{m}$etastasis from garlic #27 (ICMG-27). ICMG-27 completely blocked insulin-like growth factor-1 (IGF-1)-induced OVCAR-3 cell migration at 6 ${\mu}g/ml$. ICMG-27 completely blocked IGF-1-induced OVCAR-3 and NIH-3T3 cell migration whereas IGF-1-induced mouse embryonic fibroblast (MEF) cell migration was not affected byICMG-27. ICMG-27 inhibited all the tested IGF-1-induced cancer cell migration such as OVCAR-3, SKOV-3, and MDA-MB-231 cells. Finally, ICMG-27 could inhibit IGF-1-, lysophosphatidic acid (LPA)-, sphingosine-1-phosphate (S1P)-, leukotriene B4 (LTB4)-, and angiotensin II (AngII)-induced OVCAR-3 cell migration. These results indicate that ICMG-27 inhibits cancer cell migration by blocking essential steps in many agonists-induced cancer cell migrations. Unveiling an anti-invasive mechanism of ICMG-27 on cancer cells will provide a basis for cancer therapy.

Characterization of β-agarase from Isolated Simiduia sp. SH-4 (분리된 Simiduia sp. SH-4가 생산하는 β-agarase의 특성조사)

  • Kim, Jae-Deog;Lee, Sol-Ji;Jo, Jeong-Gwon;Lee, Dong-Geun;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.26 no.4
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    • pp.453-459
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    • 2016
  • Agarases are classified into α-agarase and β-agarase that produce agarooligosaccharides and neoagarooligosaccharides, respectively. Neoagarooligosaccharides have whitening effect of skin, delay of starch degradation, and inhibition of bacterial growth etc. Hence, the object of this study was to isolate a novel agarase producing marine bacterium and characterization of its β-agarase. A novel agar-degrading bacterium was isolated from seashore of Namhae at Gyeongnamprovine, Korea and purely cultured with Marine agar 2216 media. The isolated bacterium was identified as Simiduia sp. SH-4 after 16S rRNA gene sequencing. The enzymatic sample was obtained from culture media of Simiduia sp. SH-4. Enzymatic activity was highly increased from 20(30% relative activity) to 30℃ (100%) and decreased from 30 to 40℃(75%) and so more. Relative activity was 100% at pH 6 while those were about 91% and 59% at pH 5.0 and 7.0, respectively, meaning the enzyme possesses narrow optimal pH range. Hence, the enzyme exhibited the maximal activity with 120.4 units/l at pH 6.0 and 30℃ in 20 mM Tris-HCl buffer. Thin layer chromatography (TLC) analysis showed that Simiduia sp. SH-4 produces β-agarase, which hydrolyze agarose to produce biofunctional neoagarooligosaccharides such as neoagarotetraose and neoagarobiose. Hence, broad applications would be possible using Simiduia sp. SH-4 and its enzyme in the food industry, cosmetics and medical fields.

Isolation of a New Agar Degrading Bacterium, Maribacter sp. SH-1 and Characterization of its Agarase (신규 한천분해세균 Maribacter sp. SH-1의 분리 및 효소 특성조사)

  • Lee, Chang-Eun;Lee, Sol-Ji;Lee, Dong-Geun;Lee, Sang-Hyeon
    • Microbiology and Biotechnology Letters
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    • v.44 no.2
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    • pp.156-162
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    • 2016
  • In this study, we isolated a new agar-degrading marine bacterium and characterized its agarase. An agardegrading marine bacterium SH-1 was isolated from seawater, collected from the seashore of Namhae in Gyeongnam province, Korea, and cultured in marine agar 2216 media. It was identified as Maribacter. sp. SH-1 by phylogenetic analyses, based on 16S rRNA gene sequence. The extracellular agarase was extracted from culture media of Maribacter sp. SH-1 and characterized. Its relative activities were 56, 62, 94, 100, and 8% at 20, 30, 40, 50, and 60℃, respectively, whereas 15, 100, 60, and 21% relative activities were observed at pH 5, 6, 7, and 8, respectively. Its extracellular agarase exhibited maximum activity (231 units/l) at pH 6.0 and 50℃, in 20 mM Tris-HCl buffer. Therefore, this agarase would be applicable as it showed the maximum activity at the temperature at which the agar is in a sol state. Furthermore, the agarase activities remained over 90% at 20, 30, and 40℃ after 0.5 h exposure at these temperatures. Thin layer chromatography analysis suggested that Maribacter sp. SH-1 produces extracellular β-agarase, as it hydrolyzes agarose to produce neoagarooligosaccharides, such as neoagarohexaose (34.8%), neoagarotetraose (52.2%), and neoagarobiose (13.0%). Maribacter sp. SH-1 and its β-agarase would be useful for the production of neoagarooligosaccharides, which shows functional properties, like skin moisturizing, skin whitening, inhibition of bacterial growth, and delay in starch degradation.

Characterization of Bacillus amyloliquefaciens DA12 Showing Potent Antifungal Activity against Mycotoxigenic Fusarium Species

  • Lee, Theresa;Park, Dami;Kim, Kihyun;Lim, Seong Mi;Yu, Nan Hee;Kim, Sosoo;Kim, Hwang-Yong;Jung, Kyu Seok;Jang, Ja Yeong;Park, Jong-Chul;Ham, Hyeonheui;Lee, Soohyung;Hong, Sung Kee;Kim, Jin-Cheol
    • The Plant Pathology Journal
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    • v.33 no.5
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    • pp.499-507
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    • 2017
  • In an attempt to develop a biological control agent against mycotoxigenic Fusarium species, we isolated Bacillus amyloliquefaciens strain DA12 from soil and explored its antimicrobial activities. DA12 was active against the growth of mycotoxigenic F. asiaticum, F. graminearum, F. proliferatum, and F. verticillioides both in vitro and in planta (maize). Further screening using dual culture extended the activity range of strain DA12 against other fungal pathogens including Botrytis cinerea, Colletotrichum coccodes, Endothia parasitica, Fusarium oxysporum, Raffaelea quercus-mongolicae, and Rhizoctonia solani. The butanol extract of the culture filtrate of B. amyloliquefaciens DA12 highly inhibited the germination of F. graminearum macroconidia with inhibition rate 83% at a concentration of $31.3{\mu}g/ml$ and 100% at a concentration of $250{\mu}g/ml$. The antifungal metabolite from the butanol extract was identified as iturin A by thin layer chromatography-bioautography. In addition, volatile organic compounds produced by DA12 were able to inhibit mycelial growth of various phytopathogenic fungi. The volatile compounds were identified as 2-heptanone, 5-methyl heptanone and 6-methyl heptanone by gas chromatography-mass spectrometry (GC-MS) analysis. These results indicate that the antagonistic activity of Bacillus amyloliquefaciens DA12 was attributable to iturin A and volatile heptanones, and the strain could be used as a biocontrol agent to reduce the development of Fusarium diseases and mycotoxin contamination of crops.

PROMOTER METHYLATION OF THE CDH-13 GENE IN THE ORAL SQUAMOUS CELL CARCINOMA (구강 편평상피암종에서 CDH-13 유전자의 promoter methylation에 대한 연구)

  • Lee, Moon-Joo;Han, Se-Jin;Kim, Kyung-Wook
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.34 no.5
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    • pp.525-531
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    • 2008
  • CDH-13(T-cadherin), which is one of a kind among the 20 cadherins, can be found mainly in wall of aorta, neuron, spleen, blood vessel etc. It is also called H-cadherin. This structural difference can explain that CDH-13 is thought to play a key role in maintaining mutual relation between extra and intra-cellular environment rather than in cell adhesion. The main function of CDH-13 is to participate in blood vessel function. Additionally, it is known to regulate cell growth and cell contact inhibition. When cells are proliferating, cell surface perceives other cells so that substance such as CDH-13 can inhibit their growth or proliferation resulting in homeostasis without endless proliferation or invasion of connective tissue boundaries. However, tumor cell itself appears to be different from normal cells' growth, invasion or transmission. Therefore, it can be diagnosed that these characteristics are closely related to expression of CDH-13 in tumor cells. This study is to investigate expression of CDH-13 in SCC and its correlation with promoter methylation. 20 of tissue species for the study are excised and gathered from 20 patients who are diagnosed as SCC in department of OMS, dental hospital, dankook university. To find development of CDH-13 in each tissue samples, immunohistochemical staining, RT-PCR gene analysis and methylation specific PCR are processed. The results are as follows. 1.Immunohistochemical staining: In normal oral squamous epithelial tissue, strong expression of CDH-13 was found in cell plasma membrane of basal cell layer. On the other hand, in case of low-differentiated oral SCC, development of CDH-13 was hardly seen. 2.The development of CDH-13 gene: In 9 of samples, expression of CDH-13 gene could be seen and 2 of them showed low expression compared to the others. And rest of the 11 samples showed no expression of CDH-13 gene. 3.Methylation of CDH-13 gene: Among 9 samples which expressed CDH-13 gene, 7 of them showed unmethylation. In addition, among 11 samples without CDH-13 gene expression, 10 showed methylation. According to the results stated above, promoter methylation were found in 13 samples(65%) among 20 of oral SCC samples. In low-differentiated SCC, suppression of gene expression could be seen accompanying promoter methylation. These phenomenon of gene expression was proved by immunohistochemical investigation. Finally, for development of oral SCC, conclusions can be made that suppression of CDH-13 played a main role and suppression of gene expression was originated from promoter methylation. Considering this, it is expected that suppression of CDH-13 from promoter methylation to be utilized as a good diagnostic marker of oral SCC.

Movement of Herbicide Pretilachlor in Plants and Soils (식물체(植物體) 및 토양중(土壤中)에 있어서 제초제(際草劑) Pretilachlor의 이동특성(移動特性))

  • Ma, Sang-Yong;Moon, Young-Hee;Ryang, Hwan-Seong
    • Applied Biological Chemistry
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    • v.30 no.4
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    • pp.351-356
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    • 1987
  • This study was conducted to investigate the absorption and translocation of pretilachlor [2-chloro-2, 6-diethyl-N-(n-propoxyethyl)-acetanilide] in plants and to evaluate the mobility in soil using the $^{14}C-or$ non-labelled compound in laboratory. Rice plant(Oryza sativa L.) was very tolerant to pretilachlor. Echinochloa crus-galli P. Beauv. was completely controlled by pretilachlor at 60g a.i./10a. At the 120g a.i./l0a, growth of Cyperus serotinus Rottb. and Sagittaria pygmaea Miq. was inhibited by 75% and 25%, respectively. The growth inhibition depended on absorbed amount of $^{14}C-pretilachlor$. The greatest concentration of $^{14}C$ was found in E. crus-galli, whereas the lowest was determined in rice plant. The rate of absorption and translocation in E. curs-galli was faster than in rice plan. Pretilachlor moved to 6cm deep in sandy clay loam, clay loam and loam soils, but to 10cm in sandy loam soil. In the soils herbicide-treated layer was found 0 to 2cm profile.

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Characterization of Recombinant Bovine Sperm Hyaluronidase and Identification of an Important Asn-X-Ser/Thr Motif for Its Activity

  • Park, Chaeri;Kim, Young-Hyun;Lee, Sang-Rae;Park, Soojin;Jung, Yena;Lee, Youngjeon;Kim, Ji-Su;Eom, Taekil;Kim, Ju-Sung;Lee, Dong-Mok;Song, Bong-Suk;Sim, Bo-Woong;Kim, Sun-Uk;Chang, Kyu-Tae;Kim, Ekyune
    • Journal of Microbiology and Biotechnology
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    • v.28 no.9
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    • pp.1547-1553
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    • 2018
  • Hyaluronidases are a family of enzymes that catalyse the breakdown of hyaluronic acid, which is abundant in the extracellular matrix and cumulus oocyte complex. To investigate the activity of recombinant bovine sperm hyaluronidase 1 (SPAM1) and determine the effect of the Asn-X-Ser/Thr motif on its activity, the bovine SPAM1 open reading frame was cloned into the mammalian expression vector pCXN2 and then transfected to the HEK293 cell line. Expression of recombinant bovine hyaluronidase was estimated using a hyaluronidase activity assay with gel electrophoresis. Recombinant hyaluronidase could resolve highly polymeric hyaluronic acid and also caused dispersal of the cumulus cell layer. Comparative analysis with respect to enzyme activity was carried out for the glycosylated and deglycosylated bovine sperm hyaluronidase by N-glycosidase F treatment. Finally, mutagenesis analysis revealed that among the five potential N-linked glycosylation sites, only three contributed to significant inhibition of hyaluronic activity. Recombinant bovine SPAM1 has hyaluronan degradation and cumulus oocyte complex dispersion ability, and the N-linked oligosaccharides are important for enzyme activity, providing a foundation for the commercialization of hyaluronidase.

Characteristics of $G_{418}$-sensitive mitochondrial ATPase/ATP synthase from pleurotus florida (사철느타리버섯 중 $G_{418}$-sensitive 미토콘드리아성 ATPase/ATP synthase의 특성)

  • Kim, Jae-Woong;Kim, Dong-Hee;Lee, Jung-Bock;Lee, Sur-Koo;Min, Tae-Jin
    • Analytical Science and Technology
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    • v.5 no.4
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    • pp.477-484
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    • 1992
  • The mitochondrion was purified at 44% sucrose layer from pleurotus florida by using ultracentrifuge and sucrose density gradient method. Optimum pH and temperature of ATPase and ATP synthase were pH 7.4, $60^{\circ}C$ and pH 7.5, $57^{\circ}C$ respectively, also their Km values were determined as 11.6mM and 8.4mM. ATPase was activated at 5~6mM ATP substrate concentration, then ATP synthase was 5~10mM range ADP. ATPase/ATP synthase were $Mg^{2+}$-dependent enzyme, partially inhibited by their substrate, and then showed an none competitive inhibition pattern by $G_{418}$. Amino acid composition of ATPase/ATP synthase was as follows, hydrophobic amino acid residue was 50.5%, small residue, 56.1%, hydrogen bonding residue, 43.7% and helix breaking residue, 55.2%. Phosphatidyl choline, phosphatidyl ethanolamine and phosphatidyl glycerol were contained but not phosphatidyl inositol and phosphatidyl serine. Palmitate(51.31%), stearate(18.32%) and unsaturated fatty acids($C_{18:1}$, $C_{18:2}$ and $C_{16:1}$) were predominated.

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