• Title/Summary/Keyword: 보존과학

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Cloning and Characterization of D-xylulose Kinase from Kocuria gwangalliensis Strain SJ2 (Kocuria gwangalliensis strain SJ2에서 유래된 D-xylulose kinase 유전자의 클로닝과 특성 연구)

  • Jeong, Tae Hyug;Hwang, Tae Kyung;Seo, Yong Bae;Kim, Young Tae
    • Journal of Life Science
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    • v.25 no.5
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    • pp.507-514
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    • 2015
  • D-Xylulose is phosphorylated to D-xylulose-5-phosphate by D-xylulose kinase before it enters glycolysis via the nonoxidative pentose phosphate pathway. A gene encoding a novel D-xylulose kinase (XK) from K. gwangalliensis strain SJ2 was sequenced and expressed in E. coli. The sequence of the isolated XK gene was 1,419 bp, encoding 472 amino acids. The XK protein was more closely related to the Arthrobacter phenanthrenivorans XK than to the Bifidobacterium catenulatum one, as reflected in the sequence identity (54.9% vs. 38.7%). The XK gene was subcloned into the pCold-II expression vector. The resulting plasmid was transformed into E. coli strain BL21 (DE3) cells and the expression of the recombinant XK protein was induced by the addition of IPTG. The resulting protein was expressed as a fusion protein of approximately 48 kDa containing a N-terminal six-histidine extension that was derived from the expression vector. The expressed protein was homogenized by affinity chromatography and showed enzymatic activity corresponding to D-xylulose kinase. XK enzyme kinetic studies with D-xylulose and ATP showed a Km of 250±20 μM and 1,300±50 μM, respectively. The results obtained from this study will provide a wider knowledge base for the characterization of D-xylulose kinase at the molecular level.

Detection of Chlorotoluene and Nitrotoluene Compounds by Recombinant Microbial Biosensors (재조합 미생물 바이오센서를 이용한 chlorotoluene과 nitrotoluene 화합물의 검출)

  • Lee, Da Young;Cho, Jae Ho;Lim, Woon Ki;Shin, Hae Ja
    • Journal of Life Science
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    • v.24 no.1
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    • pp.54-60
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    • 2014
  • Aromatic hydrocarbons are toxic environmental pollutants that are detrimental to the ecosystem and human health. Among them, chlorotoluene and nitrotoluene are toxic to hydrobios and irritate the skin, eyes, and respiratory organs of humans. We herein report the development of recombinant microbial biosensors for cheap and rapid monitoring of chlorotoluene and nitrotoluene compounds. Plasmids were constructed by inserting the xylR regulatory gene for BTEX (benzene, toluene, ethylbenzene, and xylene) degradation into upstream of Po' (the DmpR activator promoter Po with the deletion of its own upstream activating sequences) or Pu (the cognate promoter of XylR)::lacZ (the ${\beta}$-galactosidase gene) and transformed into Escherichia coli $DH5{\alpha}$. In the presence of inducers, the biosensor cells immobilized in agarose developed a red color in 1-2 h due to the hydrolysis of chlorophenol red ${\beta}$-D-galactopyranoside (CPRG), a substrate of ${\beta}$-galactosidase that was expressed by the inducers. Among BTEX, high responses were specifically observed with o-, m-, p-chlorotoluene ($0.1{\mu}M-100 mM$) and o-, m-, p-nitrotoluene (0.1 mM-100 mM). Po' demonstrated higher responses than those with Pu. The biosensors immobilized in agarose showed good stability after 21 days' storage at $4^{\circ}C$, and responses in untreated wastewater spiked with chlorotoluene and nitrotoluene, suggesting they can be used to detect compounds in wastewater.

Phylogenetic Analysis of Korean Black Cattle Based on the Mitochondrial Cytochrome b Gene (mtDNA cytochrome b에 기초한 한국흑우의 계통유전학적 분석)

  • Kim, Jae-Hwan;Byun, Mi Jung;Kim, Myung-Jick;Suh, Sang Won;Kim, Young-Sin;Ko, Yeoung-Gyu;Kim, Sung Woo;Jung, Kyoung-Sub;Kim, Dong-Hun;Choi, Seong-Bok
    • Journal of Life Science
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    • v.23 no.1
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    • pp.24-30
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    • 2013
  • The purpose of this study was to identify genetic polymorphisms of the mitochondrial cytochrome b (mtDNA cyt b) gene in Korean black (KB) cattle breed and to analyze the genetic relationship between the KB and other breeds. We determined the complete sequence of the mtDNA cyt b gene in 38 KB cattle. We also analyzed their genetic diversity, and phylogenetic analysis was performed by comparison with Korean cattle (KC, called Hanwoo) and breeds from China and Japan. A nucleotide substitution was detected in the KB cattle, and two haplotypes were defined. In the neighbor-joining (NJ) tree, the haplotypes of KB were located in Bos taurus lineage with those of KC, Japanese black (JB), Yanbian and Zaosheng breeds. However, the haplotypes of Chinese breeds, excluding Yanbian and Zaosheng, were separated into B. taurus and B. indicus lineages. In the NJ tree of breeds based on Dxy genetic distances, Chinese breeds mixed with B. taurus and B. indicus lineages were located between B. indicus and B. taurus lineages. KB was contained within B. taurus lineage and was determined to be genetically more closely related to two Chinese (Yanbian and Zaosheng) breeds than to KC and JB. The haplotype distribution and the results of the phylogenetic analysis suggest that KB and KC have genetic differences in their mtDNA cyt b gene sequences.

Antioxidative Activities and Whitening Effects of Ethyl Acetate Fractions from The Immature Seeds of Abeliophyllum distichum (미선나무 미성숙 종자의 항산화 및 미백 활성)

  • Jang, Tae Won;Park, Jae Ho
    • Journal of Life Science
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    • v.27 no.5
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    • pp.536-544
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    • 2017
  • Abeliophyllum distichum Nakai is deciduous shrubs of flowering plant in Oleaceae. It is important plant resources and consists of one species in the world. Also the endemic plant of A. distichum has been protected and designed endangered plant in Korea. For this reason, study on the immature seeds of A. distichum (ADS) hasn't progressed. In the present study, we evaluated the antioxidant activity and inhibitory effects on proteins and mRNA levels were related in the whitening effect in B16F10 cells. ADS was effective for reaction oxygen species (ROS). ROS causes various diseases such as aging, inflammation, cancer, and etc. Antioxidant properties were evaluated DPPH, ABTS radical scavenging activity and Reducing power. Plants were known that contained phenolic compounds were related in antioxidant activity. Phenolic compounds were phytochemicals commonly named natural polyphenols. These are secondary metabolites of plants involved in the defense against different types of stresses. In results, ADS suppressed the expression and transcription of Tyrosinase, TRP-1, TRP-2, and Microphthalmia-associated transcription factor (MITF). Tyrosinase, tyrosinase-related protein 1 (TRP-1), tyrosinase-related protein 1 (TRP-2) are known to play an important role in melanin biosynthesis. MITF regulated the expression and transcription of Tyrosinase, TRP-1, and TRP-2. In conclusion, ADS was effective in both antioxidant activities and whitening effects. Also, they were associated with the content of phenolic compounds. We suggested that ADS can be use antioxidants and skin-whitening functional cosmetics material derived from natural plant resources.

Growth Performance and Nutrient Composition in the White-spotted Flower Chafer, Protaetia brevitarsis (Coleoptera: Scarabaeidae) Fed Agricultural By-product, Soybean Curd Cake (비지박 첨가 먹이원 급여에 따른 흰점박이꽃무지 유충의 생육과 영양성분 변화)

  • Song, Myung-Ha;Han, Moon-Hee;Lee, Seokhyun;Kim, Eun-Sun;Park, Kwan-Ho;Kim, Won-Tae;Choi, Ji-Young
    • Journal of Life Science
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    • v.27 no.10
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    • pp.1185-1190
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    • 2017
  • Insects are gaining recognition as an alternative source of protein. As a result, more and more domestic farms have begun mass rearing of edible insects. In order to produce high quality insects, studies on the development of safe and nutritious feed sources are needed. Given the cost of rearing insects, agricultural and industrial by-products are good sources for feed. The efficient utilization of these by-products can help in reducing the cost of production and preventing environmental pollution. In the current study, Citrus unshiu peel (CP), soybean curd cake (SCC), soybean oil meal (SOM), and brewers dried grain (BDG) were investigated for their effects on larval growth and development of Protaetia brevitarsis. Interestingly, the feed with 10% SCC increased larval weight by 3.5 times. For the larval period, the group of 10% SCC was significantly shorter than the control. Furthermore, minerals such as Zn, Ca, K, Mg, Na, and P were recorded to be high in 10% SCC. A total of 17 amino acids were present in 10% SCC, of which tyrosine and arginine were predominant. The heavy metal contents were very small amounts or not detected in any of the investigated groups. These findings provided a scientific basis for the utilization of soybean curd cake as a nutritional feed source to promote larval growth and produce quality insects.

A Study on Quick Quantitative Analyzing Method of Sorbic and Benzoic Acids in Doenjang by Gas Chromatography (기체크로마토그래피에 의한 시판 된장중의 소르빈산과 안식향산의 신속정량법 연구)

  • Park, Kwang-Ja;Lee, Si-Kyung;Kim, Nam-Dae;Cho, Nam-Ji
    • Korean Journal of Food Science and Technology
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    • v.31 no.2
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    • pp.285-292
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    • 1999
  • An analytical method applicable to quality control and its optimum conditions were studied for rapid and efficient analysis of sorbic and benzoic acids used as preservatives in the commercial soybean paste. In gas chromatographic analysis of sorbic and benzoic acids, the application of HP-FFAP (acid modified polyethylene) wide bore column improved the separation ability significantly. By setting the oven temperature of GC to $200^{\circ}C$, the total elapsed time for quantitative analysis was also reduced to the level required in using packed column. By extracting sorbic and benzoic acids from soybean paste with an automatic steam distillation device, the elapsed time for analysis was reduced by 80% more compared to using conventional steam distillation method. The recoveries of sorbic and benzoic acids by the automatic steam distillation were 98.1% and 99.9%, respectively. The sorbic acid was found in 3 samples of 14 commercial soybean pastes, of which contents were $466{\sim}530ppm$, while $0.3{\sim}4.4ppm$ of benzoic acid was found in all the samples.

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Separation of Highly Purified Antimicrobial Lysozyme Using Ultrafiltration and Characteristics of Membrane Fouling (한외여과 공정을 이용한 고순도 향균 Lysozyme 의 분리 및 막 침착 특성)

  • Lee, Eun-Young;Woo, Gun-Jo
    • Korean Journal of Food Science and Technology
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    • v.31 no.2
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    • pp.458-464
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    • 1999
  • The value of lysozyme as a natural food preservative is continuously increased due to its unique antimicrobial activity. To determine the optimum separation concentration among the various hen egg white protein (HEWP) concentrations (0.25, 0.5, 1.0, w/v), protein concentrations, lysozyme concentrations, specific activities (SA), and purification factors of prefiltered solution (PFS) and PM30 permeate solution (PMS) were compared. The purity of lysozyme separated at each step was analyzed and confirmed by gel permeation chromatography and electrophoresis. The fouling deposits on membrane were observed by SEM. The non-enzymatic proteins were removed over 99% by ultrafiltration (UF). The increased feed concentration did not contribute to the increase of SA. SA of PMS was 18 to 31 times higher than that of PFS. The optimum feed concentration was decided as 0.25% based on SA and purification factor. The non-enzymatic region of gel chromatogram was proved to be ovalbumin. The thickness of deposit on the UF membrane was approximately $0.9{\mu}m$ and removed by cleaning with 0.1 N NaOH. Therefore, UF using PM30 membrane was very effective to separate the antimicrobial lysozyme from various HEWPs.

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Batch Scale Storage of Garlic by Irradiation Combined with Natural Low Temperature (방사선(放射線) 조사(照射)와 자연저온(自然低溫)에 의한 마늘의 저장)

  • Cho, Han-Ok;Kwon, Joong-Ho;Byun, Myung-Woo;Yoon, Hyung-Sik
    • Korean Journal of Food Science and Technology
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    • v.16 no.1
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    • pp.66-70
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    • 1984
  • An attempt was made on the development of a commercial scale storage method of garlic by irradiation. Irradiated garlics with 50, 100 and 150 Gy were stored at natural low temperature storage room $(12{\pm}6^{\circ}C,{\;}75-85%{\;}RH)$ and the physicochemical properties during the 10 months storage were investigated. The unirradiated garlic was mostly sprouted after 8 months storage, whereas the sprouting of all irradiated groups was completely inhibited until 10 months storage, The rotting rate and weigh loss of garlic after 10 months storage were reduced by 25 to 54% at 100 Gy irradiation compared with those of an unirradiated group. The moisture content remained relatively constant during the whole storage period. The total sugar content was increased with storage period. Ascorbic acid content was also decreased until 8 months storage but its content was rapidly increased along with sprouting. Garlic was marketable after 10 months storage by 100 Gy irradiation combined with natural low temperature.

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A Study on Establishment of the Fermentation Process for Traditional Andong Sickhae (전통 안동식혜의 제조공정 확립에 관한 연구)

  • Choi, Cheong;Seog, Ho-Moon;Cho, Young-Je;Lim, Seong-Il;Lee, Woo-Je
    • Korean Journal of Food Science and Technology
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    • v.22 no.7
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    • pp.724-731
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    • 1990
  • The fermentation process of Andong Sickhae including optimal composition of the product was investigated. Through sensory evaluation and ingredient analysis of ten samples prepared in the laboratory and the samples collected from Andong region, we found that the ratio of ingredient of the product by weigh are as follow : [glutinous rice(80) : malt(50) : radish(100) : water(500) : ginger(8) : red pepper(4)]. The level of nitrogen compound turned out to be low while that of soluble protein and salt soluble protein was high. The content of total sugar and reducing sugar was found to be considerable high and among the free sugar, maltose was the highest(80%), followed by glucose and maltotriose. Activities of acid protease and saccharogenic amylase were 1.55 unit per milliliter and $12.5D^{40}_{30}^{\circ}$ respectively. Results of sensory evaluation showed that the good Andong Sickhae turned out to have well harmonized taste of flavor, sweetness and sourness while the color looked slightly red.

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On the Changes in Organic Acids of Strawberry in Air with Different $CO_2$ Concentration (공기중에 혼합된 탄산가스 농도에 따른 저장딸기의 유기산 변화에 관하여)

  • Kim, Dong-Man;Kim, Kil-Hwan;Kim, Chang-Sik
    • Korean Journal of Food Science and Technology
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    • v.18 no.1
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    • pp.71-76
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    • 1986
  • Changes in pH, titratable acidity, some organic acids and brix of the strawberry stored at the different levels of $CO_2$, in air were measured. The changing trends in pH and the acidity of the strawberries were not clear among the storage treatments, and pH of the strawberries in all the treatments was increased(r=0.9615) by contrast to decrease in the acidity(r=-0.8483). Ascorbic acid content of the strawberries at control lot was higher than the contents of the CA stored for the initial 2 weeks after storage. This trend was reversed after that period and the strawberries in air with 30% $CO_2$, showed the highest retention rate, 28% of the initial content after 5 weeks. Among the analyzed organic acids, malic acid was gradually decreased in all the treatments during storage and the decreasing rates of the strawberries in air with 30% $CO_2$, was somewhat stupider than those in the others. Pyruvic acid was reduced in the control lot during storage but drastically increased in the strawberries stored at the CA storage conditions between the 2nd and the 3rd week after storage, and then it was decreased.

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