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Improvement in Antagonistic Ablility of Antagonistic Bacterium Bacillus sp. SH14 by Transfer of the Urease Gene. (Urease gene의 전이에 의한 길항세균 Bacillus sp. SH14의 길항능력 증가)

  • 최종규;김상달
    • Microbiology and Biotechnology Letters
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    • v.26 no.2
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    • pp.122-129
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    • 1998
  • It were reported that antifungal mechanism of Enterobacter cloacae is a volatile ammonia that produced by the strain in soil, and the production of ammonia is related to the bacterial urease activity. A powerful bacterium SH14 against soil-borne pathogen Fusarium solani, which cause root rot of many important crops, was selected from a ginseng pathogen suppressive soil. The strain SH14 was identified as Bacillus subtilis by cultural, biochemical, morphological method, and $API^{circledR}$ test. From several in vitro tests, the antifungal substance that is produced from B. subtilis SH14 was revealed as heat-stable and low-molecular weight antibiotic substance. In order to construct the multifunctional biocontrol agent, the urease gene of Bacillus pasteurii which can produce pathogenes-suppressive ammonia transferred into antifungal bacterium. First, a partial BamH I digestion fragment of plasmid pBU11 containing the alkalophilic B. pasteurii l1859 urease gene was inserted into the BamH I site of pEB203 and expressed in Escherichia coli JM109. The recombinant plasmid was designated as pGU366. The plasmid pGU366 containing urease gene was introduced into the B. subtilis SH14 with PEG-induced protoplast transformation (PIP) method. The urease gene was very stably expressed in the transformant of B. subtilis SH14. Also, the optimal conditions for transformation were established and the highest transformation frequency was obtained by treatment of lysozyme for 90 min, and then addition of 1.5 ${mu}g$/ml DNA and 40% PEG4000. From the in vitro antifungal test against F. solani, antifungal activity of B. subtilis SH14(pGu366) containing urease gene was much higher than that of the host strain. Genetical development of B. subtilis SH14 by transfer of urease gene can be responsible for enhanced biocontrol efficacy with its antibiotic action.

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Biochemical Characteristics for the Cofactor Free Mutant of Yeast Homocysteine Catalyzing Enzyme, Cystathionine ${\beta}$-Synthase (조효소를 함유하지 않는 효모의 Homocysteine 분해효소, Cystathionine ${\beta}$-Synthase의 생화학적 특성)

  • Jhee, Kwang-Hwan;Cho, Hyun-Nam;Yang, Seun-Ah;Lee, In-Seun
    • Microbiology and Biotechnology Letters
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    • v.35 no.3
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    • pp.196-202
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    • 2007
  • Mutations in the cystathionine ${\beta}$-synthase (CBS) gene cause homocystinuria, the most frequent inherited disorder in sulfur metabolism. CBS is the unique enzyme using both heme and pyridoxal 5-phosphate (PLP) for activity. Among the reported 140 mutations, one of the most common disease-causing alterations in human CBS is G307S mutation. To investigate the pathogenic mechanism of G307S by spectroscopic methods, we engineered the full length and the truncated G247S mutation of yeast CBS that is corresponding mutation to human G307S. Yeast CBS does not contain heme and thus gives a merit to study the spectroscopic properties. The UV-visible spectra of the purified full length and the truncated G247S yeast CBSs showed the total absence of PLP in the protein. The absence of PLP in G247S mutation was also confirmed by the PLP-cyanide adduct formation experiment, which was conducted by the incubation of the purified enzyme with KCN. The adducts were detected using a circular dichroism (CD) and a spectrofluorimeter. Radio isotope activity assay of full length and truncated G247S proteins also gave no activity. Our yeast G247S mutation data suggested that G307S might make the distortion of the active site so that cofactor PLP and substrate can not fit inside the active site. Our yeast CBS study addressed the reason why the G307S mutation in human CBS makes the enzyme inactive that consequently leads to severe clinical phenotype.

Biochemical Characterization of a Novel Thermostable Esterase from the Metagenome of Dokdo Islets Marine Sediment (독도 심해토 메타게놈 유래 신규 내열성 에스테라아제의 생화학적 특성규명)

  • Lee, Chang-Muk;Seo, Sohyeon;Kim, Su-Yeon;Song, Jaeeun;Sim, Joon-Soo;Hahn, Bum-Soo;Kim, Dong-Hern;Yoon, Sang-Hong
    • Microbiology and Biotechnology Letters
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    • v.45 no.1
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    • pp.63-70
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    • 2017
  • A functional screen of 60,672 fosmid metagenomic clones amplified from marine sediment obtained from the Dokdo islets in Korea identified the gene EstES1, whose product, EstES1, displayed lipolytic properties on tributyrin-supplemented media. EstES1 is a 576 amino acid protein with a predicted molecular weight of 59.4 kDa including 37 N-terminal leader amino acids. EstES1 exhibited the highest sequence similarity (44%) to a carboxylesterase found in Haliangium ochraceum DSM14365. Phylogenetic analysis indicated that EstES1 belongs to a currently uncharacterized family of lipases. Within the conserved domain, EstES1 retains the catalytic triad that consists of the consensus penta-peptide motif, GESAG. EstES1 demonstrated a broad substrate specificity toward the long acyl group of ethyl esters (C2-C12), and its optimal activity was recorded toward p-Nitrophenyl butyrate (C4) at pH 9.0 and $40^{\circ}C$ (specific activity of 255.4 U/mg). The enzyme remained stable in the ranges of $60-65^{\circ}C$ and pH 9.0-10.5 and in the presence of methanol, ethanol, isopropanol, and dimethyl sulfoxide. Therefore, EstES1 has potential for use in industrial applications involving high temperature, organic solvents, and/or alkaline conditions.

A Study on the Moisturizing Effect and Preparation of Liquid Crystal Structures Using Sucrose Distearate Emulsifier (슈크로오스디스테아레이트를 사용한 액정구조의 생성과 보습효과에 관한 연구)

  • Kwak, Myeong-Heon;Kim, In-Young;Lee, Hwan-Myung;Park, Joo-Hoon
    • Journal of the Korean Applied Science and Technology
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    • v.33 no.1
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    • pp.1-12
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    • 2016
  • This study is to make the liquid crystalline structure using sucrose distearate (Sucro-DS) emulsifier to create the hydrophilic type oil-in-water (O/W) emulsion, the droplets of the emulsion having a structure of a multi-lamellar structure. We have studied the physicochemical properties of Sucro-DS using those techniques. And it has been studied in the emulsion performance. In order to form the liquid crystalline structure applying 3 wt% of Sucro-DS, 5 wt% of glycerin, 5 wt% of squalane, 5 wt% of capric/caprylic triglyceride, 3wt% of cetostearyl alcohol, 1wt% of glyceryl mono-stearate, 78 wt% of pure water in mixture having the lamellar structure of stable multi-layer system was found to formed. By applying them, they were described how to create an unstable active material encapsulated cream. Further, the moisturizing cream was studied using this technique. It reported the results to the skin improvement effect by the human clinical trials. The pH range to produce a stable liquid crystal phase using a Sucro-DS was maintained in 5.2~7.5. In order to increase the stability of the liquid crystal, it was when behenyl alcohol containing 3 wt%, the hardness at this time was 13 kg/mm,min. Viscosity of the same amount was 25,000mPas/min. After a test for the effects of the emulsions, the concentration of 6 wt% Sucro-DS is that was appropriate, the particle size of the liquid crystal was 4~6mm. It was observed through a microscope analysis, reliability of the liquid crystal changes for 3 months was found to get stable at each $4^{\circ}C$, $25^{\circ}C$ and $45^{\circ}C$. In clinical trial test, before applying a moisturizing effect it was $13.4{\pm}7%$. Moisturizing cream liquid crystal was not formed in $14.5{\pm}5%$. Therefore, applying than ever before could see the moisture about 8.2% was improved. On the other hand, it was the moisturizing effect of the liquid cream is $19.2{\pm}7%$. The results showed that 43.3% improvement than that previously used. Applications fields, Sucro-DS emulsifier used liquid cream, lotion, eye cream and a variety of formulations can be developed, as well as the cosmetics industry is expected to be wide fields in the application of the external preparation for skin emulsion technology in the pharmaceutical industry and pharmaceutical industry.

Regeneration Processes of Nutrients in the Polar Front Area of the East Sea II. Distribution of Particulate Organic Carbon and Nitrogen in Winter, 1995 (동해 극전선의 영양염류 순환과정 II. 1995년 동계 입자태 유기탄소 및 유기질소의 분포)

  • YANG Han-Soeb;MOON Chang-Ho;OH Seok-Jin;LEE Haeng-Pil
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.30 no.3
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    • pp.442-450
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    • 1997
  • The chemical properties of water masses were investigated at 33 stations of the southeastern last Sea in February, 1995 on board R/V Tam-Yang. The water masses were not clearly distinguished due to the vortical mixing in winter. However, on the basis of the T-S and $T-O_2$ diagrams, water masses in the study area were divided into five groups (Type I, Type II, Type III, Type IV, Type V). (1) $>9.0^{\circ}C,\;>34.35\;psu,\;5.08\~5.60m\ell/\ell$ at Type I, (2) $6.0\~9.0^{\circ}C,\;34.15\~34.35\;psu,\;5.60\~5.90\;m\ell/\ell$ at Type II, (3) $4.0\~6.0^{\circ}C,\;34.00\~34.15\;psu,\;>5.90m\ell/\ell$ at Type III, (4) $1.5\~4.0^{\circ}C,\;34.00\~34.05\;psu,\;5.40\~5.90\;m\ell/\ell$ at Type IV, (5) $<1.5^{\circ}C,\;34.05\~34.07\;psu,\;4.80\~5.40\;m\ell/\ell$ at Type V. In the vertical profiles of nutrients, the concentrations were very low in the surface layer and increased rapidly with depth. The highest concentrations occurred in Type IV, while the concentrations in Type I were the lowest. The N/P ratios were less than Redfield ratio, indicating that nitrogenous nutrients were the limiting factor tor phytoplankton growth. The concentrations of POC and PON were in the range of $0.49\~20.03\;{\mu}g-at/\ell\;and\;0.09\~5.34\;{\mu}g-at/\ell$, respectively. The relatively high concentration occured in the surface layer of inner shore, showing that the concentration at each water mass followed the order Type I > Type II > Type III > Type IV > Type V, respectively. The C:N ratio in particulate organic matter was lower than the values reported in other region due to relatively high concentrations of PON in the study area. Relatively high ratios of POC to chlorophyll $\alpha$ during the study periods indicate that non-living detritus comparised most of the POC in the study area.

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Regeneration Processes of Nutrients in the Polar Front Area of the East Sea III. Distribution Patterns of Water Masses and Nutrients in the Middle-Northern last Sea of Korea in October, 1995 (동해 극전선역의 영양염류 순환 과정 III. 1995년 10월 동해 중부 및 북부 해역의 수괴와 영양염의 분포)

  • CHO Hyun-Jin;MOON Chang-Ho;YANG Han-Seob;KANG Won-Bae;LEE Kwang-Woo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.30 no.3
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    • pp.393-407
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    • 1997
  • A survey of biological and chemical characteristics in the middle-northern East Sea of Korea was carried out at 28 stations in October, 1995 on board R/V Tam-Yang. On the basis of the vertical profiles of temperature, salinity and dissolved oxygen, water masses in the study area were divided into 5 major groups; (1) Low Saline Surface Water (LSSW), (2) Tsushima Surface Water (TSW), (3) Tsushima Middle Water (TMW), (4) North Korean Cold Water (NKCW), (5) last Sea Porper Water (ESPW). Other 4 mixed water masses were also observed. It is highly possible that the LSSW which occured at depths of $0\~30m$ in the most southern part of the study area is originated from the Yangtze River (Kiang) of China due to very low salinity $(<32.0\%_{\circ})$ relatively high concentration of dissolved silicate and no sources of freshwater input into that area. Oxygen maximum layer in the vertical profile was located near surface at northern cold waters and became deeper at the warm southern area. Oxygen minimum layer af depths $50\~100m$, which is TMW, were found in only southern area. In the vortical profiles of nutrients, the concentrations were very low in the surface layer and increased drammatically near the thermocline. The highest concentration occurred in the ESPW. The relatively low value of Si/P ratio in the ESPW (13.63) compared to other reports in the East Sea was due to continuous increase of P with depth as well as Si. The N : P ratio was about 6.92, showing that nitrogenous nutrient is the limiting factor for phytoplankton growth. The exponential relationship between Si and P, compared to the linear relationship between N and P, indicates that nitrate and phosphate have approximately the same regenerative pattern, but silicate has delayed regenerative pattern.

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A Study on Sea Water and Ocean Current in the Sea Adjacent to Korea Peninsula -III. Chemical Characteristics of Water Masses in the Polar Front Area of the Central Korean East Sea- (한반도 근해의 해류와 해수특성 -III. 한국 동해 중부 극전선역에 출현하는 수괴의 화학적 특성-)

  • YANG Han-Soeb;KIM Seong-Soo;KANG Chang-Geun;CHO Kyu-Dae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.24 no.3
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    • pp.185-192
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    • 1991
  • The vertical distribution and chemical characteristics of water masses were measured along two south-north transects in the polar front region of the central Korean East Sea. In February, a thermocline was present at depth between 50m and loom at the southern sites of a landward A-transect, and its depth was gradually deepened northward. At an outside B-transect, a thermocline was observed at significantly deep depth of 300m to 400m at two northern stations(Stn. 10 and 11), though the depth of the southward stations was nearly identical to that at the northward stations on a A-transect. In September, there were vertically more various water masses, i.e. the Tsushima Warm surface water(TWSW) or more than $20^{\circ}C$, the Tsushima Middle water(TMW) with a range of $12{\~}17^{\circ}C$, the North Korea Cold Water(NKCW) with $1{\~}7^{\circ}C$ temperature, the Japan Sea Proper Water(JSPW) of less than $1^{\circ}C$, and the mixed water. The North Korea Cold Water could be distinguishable from the other waters, especially from the mixed water of the Tsushima Middle Water and the Japan Sea Proper Water by the pattern of $T-O_2$ diagram. For instance, the North Korea Cold Water had higher oxygen by $1{\~}2ml/l$ than those in the mixed water, although both the two water masses ranged $1{\~}7^{\circ}C$ in water temperature. AOU value was the highest in the JSPW and the lowest in the TWSW. Also, AOU indicated a nearly linear and negative correlation with water temperature. However, AOU data for two masses, the NKCW and the TMW, in September departed remarkably from a regression line. Moreover, the ratio of $$\Delta P/\Delta AOU)$ in September was about $0.45{\mu}g-at/ml$ and higher than the value observed in the open sea. This high value could be elucidated by two factors; intrusion of the NKCW with high oxygen and molecular diffusion of dissolved oxygen from the surface into the lower layer. AOU would be a useful tracer for water masses in the polar front area of the Korean East Sea.

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EFFECT OF LIGHT IRRADIATION MODES ON THE MARGINAL LEAKAGE OF COMPOSITE RESIN RESTORATION (광조사 방식이 복합레진 수복물의 변연누출에 미치는 영향)

  • 박은숙;김기옥;김성교
    • Restorative Dentistry and Endodontics
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    • v.26 no.4
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    • pp.263-272
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    • 2001
  • The aim of this study was to investigate the influence of four different light curing modes on the marginal leakage of Class V composite resin restoration. Eighty extracted human premolars were used. Wedge-shaped class Y cavities were prepared on the buccal surface of the tooth with high-speed diamond bur without bevel. The cavities were positioned half of the cavity above and half beyond the cemento-enamel junction. The depth, height, and width of the cavity were 2 mm, 3 mm and 2 mm respectively. The specimens were divided into 4 groups of 20 teeth each. All the specimen cavities were treated with Prime & Bond$^{R}$ NT dental adhesive system (Dentsply DeTrey GmbH, Germany) according to the manufacturer's instructions and cured for 10 seconds except group VI which were cured for 3 seconds. All the cavities were restored with resin composite Spectrum$^{TM}$ TPH A2 (Dentsply DeTrey GmbH, Germany) in a bulk. Resin composites were light-cured under 4 different modes. A regular intensity group (600 mW/${cm}^2$, group I) was irradiated for 30 s, a low intensity group (300 mW/${cm}^2$, group II) for 60 s and a ultra-high intensity group (1930 mW/${cm}^2$, group IV) for 3 s. A pulse-delay group (group III) was irradiated with 400 mW/${cm}^2$ for 2 s followed by 800 mW/${cm}^2$ for 10 s after 5 minutes delay. The Spectrum$^{TM}$ 800 (Dentsply DeTrey GmbH, Germany) light-curing units were used for groups I, II and III and Apollo 95E (DMD, U.S.A.) was used for group IV. The composite resin specimens were finished and polished immediately after light curing except group III which were finished and polished during delaying time. Specimens were stored in a physiologic saline solution at 37$^{\circ}C$ for 24 hours. After thermocycling (500$\times$, 5-55$^{\circ}C$), all teeth were covered with nail varnish up to 0.5 mm from the margins of the restorations, immersed in 37$^{\circ}C$, 2% methylene blue solution for 24 hours, and rinsed with tap water for 24 hours. After embedding in clear resin, the specimens were sectioned with a water-cooled diamond saw (Isomet$^{TM}$, Buehler Co., Lake Bluff, IL, U.S.A.) along the longitudinal axis of the tooth so as to pass the center of the restorations. The cut surfaces were examined under a stereomicroscope (SZ-PT Olympus, Japan) at ${\times}$25 magnification, and the images were captured with a CCD camera (GP-KR222, Panasonic, Japan) and stored in a computer with Studio Grabber program. Dye penetration depth at the restoration/dentin and the restoration/enamel interfaces was measured as a rate of the entire depth of the restoration using a software (Scion image, Scion Corp., U.S.A.) The data were analysed statistically using One-way ANOVA and Tukey's method. The results were as follows : 1. Pulse-Delay group did not show any significant difference in dye penetration rate from other groups at enamel and dentin margins (p>0.05) 2. At dentin margin, ultra-high intensity group showed significantly higher dye penetration rate than both regular intensity group and low intensity group (p<0.05). 3. At enamel margin, there were no statistically significant difference among four groups (p>0.05). 4. Dentin margin showed significantly higher dye penetration rate than enamel margin in all groups (p<0.05).

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MICROLEAKAGE OF THE CLASS V CAVITY ACCORDING TO RESTORATION SITE AND CAVITY SIZE USING SEM AND THREE-DIMENSIONAL RECONSTRUCTION TECHNIQUES (SEM과 3차원 재구성법을 이용한 수복면의 위치와 와동 크기에 따른 미세누출도 분석)

  • Yang, In-Seo;Shin, Dong-Hoon
    • Restorative Dentistry and Endodontics
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    • v.30 no.2
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    • pp.112-120
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    • 2005
  • This study was done to evaluate whether there were any differences in microleakage of class V composite restorations according to restoration site and cavity size. Total sixty-four restorations were made in molar teeth using Esthet-X. Small ($2\;{\times}\;2\;{\times}\;1.5\;mm$) and large ($4{\times}2{\times}1.5\;mm$) restorations were made at the buccal/lingual surface and the proximal surface each. After 1,000 times of thermocycling ($5^{\circ}\;-\;55^{\circ}C$), resin replica was made and the percentage of marginal gap to the whole periphery of the restoration was estimated from SEM evaluation. Thermocycled tooth was dye penetrated with $50\%$ silver nitrate solution. After imbedding in an auto-curing resin, it was serially ground with a thickness of 0.25 mm. Volumetric microleakage was estimated after reconstructing three dimensionally. Two-way ANOVA and independent T-test for dye volume, Mann-Whitney U test for the percentage of marginal gap, Spearman's rho test for the relationship between two techniques were used, The results were as follows : 1. The site and size of the restoration affected on the microleakage of restoration. Namely, much more leakage was seen in the proximal and the large restorations rather than the buccal/lingual and the small restorations. 2. Close relationship was found between two techniques (Correlation coefficient = 0.614/ P = 0.000). Within the limits of this study, it was noted that proximal and the large restorations leaked more than buccal/lingual and the small restorations. Therefore, it should be strictly recommended large exposure of margins should be avoided by reducing unnecessary tooth reduction.

Study on the Processing and Compositions of Salted and Dried Mullet Roe (영암산 염건 숭어알의 가공과 조성에 관한 연구)

  • Joe, Sang-June;Rhee, Chong-Ouk;Kim, Dong-Youn
    • Korean Journal of Food Science and Technology
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    • v.21 no.2
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    • pp.242-251
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    • 1989
  • The salted-dried mullet(Mugil japonicus) roe is a kind of traditional food particulary in the area of Young-am gun, Chunnam province. This study was conducted to conform the scientific processing conditions and to evaluate the nutritional quality and changes of major components during storage times. The manufacturing method was that the fresh roe was salted for about 20 hours for the preparation of salted-dried roe, washed by clean waters, drained, shaped a flat piece with 1.2cm thickness by pressing, and spreaded sesame oils on the surface of the salted roe periodically during wind drying for 20 days. The dried roe was blanched in heated water$(80^{\circ}C/3min)$ and packaged the dried product for storages. The fractional compositions of free lipid of wind dried roe were 40% of neutral lipids, 12% of glycolipids and 9% of phospholipids and those of bound lipids were 13% of neutral lipids. 10% of glycolipids and 13% of phospholipids respectively. The major fatty acids of the roe were $C_{16:0}$, $C_{18:0}$, $C_{18:1}$, $C_{18:2}$ and $C_{20:0}$ which was consisted of free and bound lipids in wind drying method during processing and storages. Total amino acids were 99.87g/100g and major amino acids were Glu, Pro, Leu, Lys and CySH and the protein score was average 155% and the chemical score was average 109%. Free amino acids was 1,376mg% that had 50.61% of Pro and the major kinds of those were Tyr and CySH.

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