• Title/Summary/Keyword: Cell Characterization

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Preparation and Characterization of Enzymatic Oyster Hydrolysates-added Yogurt (굴 효소 가수분해물 첨가 요구르트의 제조 및 특성)

  • Chung, In-Kwon;Kim, Hye-Suk;Kang, Kyung-Tae;Choi, Jong-Duck;Heu, Min-Soo;Kim, Jin-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.7
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    • pp.926-934
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    • 2006
  • The base for preparing oyster hydrolysate-added yogurt was consisted of whole milk (1,000 mL), skim milk (44.05 to 42.05 g), enzymatic oyster hydrolysates powder (OHP, 0 to 2.0 g) and pectin. The yogurt base was fermented with 7 kinds of starter cultures (3% based on yogurt volume), such as Lactobacillus acidophilus, lactobacillus bulgaricus, lactobacillus casei, Lactobacillus fermentum, Lactobacillus pentosus, Streptcoccus thermophilus and the mixed starters (L. bulgaricus and S. thermophilus) at optimal temperature. Processing condition and quality characteristics of the yogurt were evaluated by analyzing pH, titratable acidity, viscosity, viable cell count, functional properties and sensory evaluation. The results suggested that the optimal conditions for preparing the good quality yogurt revealed the mixed starters (L. bulgaricus and S. thermophilus) for starter culture, 1.0 g of 3 kDa hydrolysate for amount, and 5.5 hrs for fermentation time. The good quality yogurt showed 4.31 for pH, 1.07% for titratable acidity, 469 cps for viscosity and $4.9{\times}10^8\;CFU/mL$ for viable cell count. The hydrolysate-added yogurt was 2 times higher in ACE inhibitory and antioxidant activities than commercial yogurt, and kept good quality during storage of 15 days at $5^{\circ}C$.

Characterization of a cDNA Encoding Transmembrane Protein 258 from a Two-spotted Cricket Gryllus bimaculatus (쌍별귀뚜라미(Gryllus bimaculatus)의 GbTmem258 cDNA 클로닝과 발현분석)

  • Kisang Kwon;Honggeun Kim;Hyewon Park;O-Yu Kwon
    • Journal of Life Science
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    • v.33 no.10
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    • pp.828-834
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    • 2023
  • The cDNA that encodes transmembrane protein 258 (Tmem258) was cloned from Gryllus bimaculatus and named GbTmem258. This protein comprises 80 amino acids, has no N-glycosylation site, and contains five potential phosphorylation sites at two serines, two threonines, and one tyrosine. The predicted molecular mass of GbTmem258 is 9.06 kDa, and its theoretical isoelectric point is 5.5. The tertiary structure of GbTmem258 was predicted using the available secondary structure information, which suggests the presence of alpha helices (52.5%), random coils (22.5%), extended strands (16.25%), and beta turns (8.75%). Homology analysis revealed that GbTmem258 exhibits high similarity at the amino-acid level to Tmem258 found in other species. The effect of starvation and refeeding on GbTmem258 mRNA expression was also examined in this study. It was found that GbTmem258 mRNA expression in the hindgut progressively increased throughout the starvation period, peaking at almost 1.5 times the control level after six days of starvation. However, refeeding for one to two days after the six-day starvation period restored GbTmem258 mRNA expression to the control level. In fat body, GbTmem258 mRNA expression was almost 3-fold higher during starvation compared to the control level. Refeeding for one to two days after the six-day fast resulted in a decline in the expression to about a 2.5-fold increase over the control level. Throughout the starving and refeeding periods, no other tissues showed any discernible alterations in GbTmem258 mRNA expression.

Mineralogical Characterization of the Chuncheon Nephrite: Mineral Facies, Mineral Chemistry and Pyribole Structure (춘천 연옥 광물의 광물학적 특성 : 광물상, 광물 화학 및 혼성 격자 구조)

  • Noh, Jin Hwan;Cho, Hyen Goo
    • Journal of the Mineralogical Society of Korea
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    • v.6 no.2
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    • pp.57-79
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    • 1993
  • Chuncheon nephrite, which was formed by the polymetasomatic alteration of dolomitic marble, can be classified into pale green, green, dark green, and grey types on the basis of their occurrence, mineralogical and textural characteristics. The nephrites consist obiefly of fibrous or hairlike(length/width ratio>10) cryptocrystalline(crystal width < $2{\mu}m$) tremolite, and include less amounts of micro-crystalline diopside, calcite, clinochlore, and sphene as impurities. The oriented and rather curved crystal aggregate, of nephritic tremolite are densely interwoven, resulting in a massive-fibrous texture which may explain the characteristic toughness of nephritic jade. The characteristic greenish color of the nephrite may be preferably related to Fe rather than Cr and Ni. However, the variation of color and tint in the Chuncheon nephrite also depends on the mineralogical and textural differences such as crystallinity, texture, and impurities. The chemical composition of the nephritic tremolite is not stoichiometric and rather dispersed especially in the abundances of Al, Mg, and Ca. Al content and Mg/Ca ratio for the nephritic tremolite are slightly increased with deepening in greenish color of the nephrite. Fe content in the nephritic tremolite is generally very low, but comparatively richer in the dark green nephrite. In nephritic tremolite, wide-chain pyriboles are irregularly intervened between normal double chains, forming a chain-width disorder. Most nephritic tremolites in the Chuncheon nephrite show various type of chain-width defects such as triple chain(jimthompsonite), quintuple chain (chesterite), or sometimes quadruple chain in HRTEM observations. The degree of chain-width disorder in the nephritic tremolite tends to increase with deepening in greenish color. Triple chain is the most common type, and quadruple chain is rarely observed only in the grey nephrite. The presence of pyribole structure in the nephritic tremolite is closely related to the increase of Al content and Mg/Ca ratio, a rather dispersive chemical composition, a decrease of relative intensity in (001) XRD reflection, and an increase in b axis dimension of unit cell. In addition, the degree and variation of chain-width disorder with nephrite types may support that an increase of metastability was formed by a rapid diffusion of Mg-rich fluid during the nephrite formation.

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Characterization of a Psychrophilic Metagenome Esterase EM2L8 and Production of a Chiral Intermediate for Hyperlipemia Drug (메타게놈유래의 저온성 에스터라제 EM2L8의 효소적 특성과 이를 활용한 고지혈증 치료제 키랄소재의 생산)

  • Jung, Ji-Hye;Choi, Yun-Hee;Lee, Jung-Hyun;Kim, Hyung-Kwoun
    • Microbiology and Biotechnology Letters
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    • v.37 no.2
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    • pp.118-124
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    • 2009
  • Esterase EM2L8 gene isolated from deep sea sediment was expressed in Escherichia coli BL21 (DE3) and the esterase activity of the cell-free extract was assayed using p-nitrophenyl butyrate-spectrophotometric method. Its optimum temperature was $40-45^{\circ}C$ and 45% activity of the maximum activity was retained at $15^{\circ}C$. The activation energy at $15-45^{\circ}C$ was calculated to be 4.9 kcal/mol showing that esterase EM2L8 was a typical cold-adapted enzyme. Enzyme activity was maintained for 6 h and 4 weeks at $30^{\circ}C$ and $4^{\circ}C$, respectively. When each ethanol, methanol, and acetone was added to the reaction mixture to 15% concentration, enzyme activity was maintained. In the case of DMSO, enzyme activity was kept up to 40% concentration. (S)-4-Chloro-3-hydroxy butyric acid is a chiral intermediate for the synthesis of Atorvastatin, a hyperlipemia drug. When esterase EM2L8 (40 U) was added to buffer solution (1.2 mL, pH 9.0) containing ethyl-(R,S)-4-chloro-3-hydroxybutyrate (38 mM), it was hydrolyzed into 4-chloro-3-hydroxy butyric acid with a rate of $6.8\;{\mu}mole/h$. The enzyme hydrolyzed (S)-substrate more rapidly than (R)-substrate. When conversion yield was 80%, e.e.s value was 40%. When DMSO was added, hydrolysis rate increased to $10.4\;{\mu}mole/h$. The plots of conversion yield vs e.e.s in the presence or absence of DMSO were almost same, implying that the reaction enantioselectivity was not changed by the addition of DMSO. Taken together, esterase EM2L8 had high activity and stability at low temperatures as well as in various organic solvents/aqueous solutions. These properties suggested that it could be used as a biocatalyst in the synthesis of useful pharmaceuticals.

Characterization and Formation Mechanisms of Clogging Materials in Groundwater Wells, Mt. Geumjeong Area, Busan, Korea (부산 금정산 일대 지하수공내 공막힘 물질의 특징과 형성원인)

  • Choo, Chang-Oh;Hamm, Se-Yeong;Lee, Jeong-Hwan;Lee, Chung-Mo;Choo, Youn-Woo;Han, Suk-Jong;Kim, Moo-Jin;Cho, Heuy-Nam
    • The Journal of Engineering Geology
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    • v.22 no.1
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    • pp.67-81
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    • 2012
  • The physical, chemical, and biological properties of clogging materials formed within groundwater wells in the Mt. Geumjeong area, Busan, Korea, were characterized. The particle size distribution (PSD) of clogging materials was measured by a laser analyzer. XRD, SEM, and TEM analyses were performed to obtain mineralogical information on the clogging materials, with an emphasis on identifying and characterizing the mineral species. In most cases, PSD data exhibited an near log-normal distribution; however, variations in frequency distribution were found in some intervals (bi-or trimodal distributions), raising the possibility that particles originated from several sources or were formed at different times. XRD data revealed that the clogging materials were mainly amorphous ironhydroxides such as goethite, ferrihydrite, and lapidocrocite, with lesser amounts of Fe, Mn, and Zn metals and silicates such as quartz, feldspar, micas, and smectite. Reddish brown material was amorphous hydrous ferriciron (HFO), and dark red and dark black materials were Fe, Mn-hydroxides. Greyish white and pale brown materials consisted of silicates. SEM observations indicated that the clogging materials were mainly HFO associated with iron bacteria such as Gallionella and Leptothrix, with small amounts of rock fragments. In TEM analysis, disseminated iron particles were commonly observed in the cell and sheath of iron bacteria, indicating that iron was precipitated in close association with the metabolism of bacterial activity. Rock-forming minerals such as quartz, feldspar, and micas were primarily derived from soils or granite aquifers, which are widely distributed in the study area. The results indicate the importance of elucidating the formation mechanisms of clogging materials to ensure sustainable well capacity.

Physicochemical Properties of Colostrum by Milking Time of Gyeonggi Province (경기지역의 착유회수에 따른 초유의 이화학적 특성)

  • Jeong, Seok-Geun;Ham, Jun-Sang;Kim, Dong-Hun;Ahn, Chong-Nam;Chae, Hyun-Seok;You, Young-Mo;Jang, Ae-Ra;Kwon, Il-Kyung;Lee, Seung-Gyu
    • Food Science of Animal Resources
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    • v.29 no.4
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    • pp.445-456
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    • 2009
  • Colostrum samples were collected from 36 dairy farms in Gyeonggi-do and one dairy farm in the National Institute of Animal Science (NIAS) for testing. Colostrum samples were analyzed for phisycochemicals (specific gravity, pH, titratable acidity), general components (fat, protein, lactose, total solid, solid non-fat (SNF)), fatty acids, amino acids, minerals, microflora, somatic cells, and Ig (Immunoglobulin). The first colostrum revealed the following data: fat contents were $6.16{\pm}2.39%$, proteins were $14.78{\pm}4.30%$, lactose $2.57{\pm}0.77%$, total solid $24.28{\pm}4.36%$, and SNF $18.12{\pm}4.08%$, whereas the 2nd (or $12^{th}$) colostrum revealed $5.56{\pm}1.76%$ fat, $3.46{\pm}0.41%$ proteins, $4.19{\pm}0.43%$ lactose, $13.90{\pm}1.76%$ total solid, and $8.34{\pm}0.81%$ SNF. Also, the first colostrum revealed the contents of major amino acids as 0.89% aspartic acid, 0.71% threonine, 0.86% serine, 1.75% glutamic acid, 0.64% valine, 0.95% leucine, 0.83% lysine, and 0.95% proline, and those in the 10th colostrum were 0.25% aspartic acid, 0.15% threonine, 0.19% serine, 0.59% glutamic acid, 0.19% valine, 0.35% leucine, 0.31% lysine, and 0.34% proine. Major amino acid contents rapidly decreased as milking times increased. In the first colostrum, the following mineral contents were observed: there were 2,168 ppm in Ca, 1,959 ppm in P, 914 ppm in K, 761 ppm in Na, 287 ppm in Mg, 1.7 ppm in Fe, 14.3 ppm in Zn, and 1.0 ppm in Cu; while in the 10th colostrum, the following ppm contents were 1,389 in Ca, 1,323 in P, 838 in K, 427 in Na, 131 in Mg, 1.0 in Fe, 4.7 in Zn, and 1.3 in Cu. The mineral contents in a colostrum rapidly decreased as milking times increased.

Isolation of Synthetic Detergent Decomposing Microorganisms in Wastewater and Synthetic Detergent Decomposition Characterization of the Microorganisms (폐하수중 합성세제분해균의 분리(分離) 및 합성세제 (ABS) 분해특성(分解特性))

  • Lee, Hong-Jae;Heo, Jong-Soo;Cho, Ju-Sik;Han, Mun-Gyu;Choi, Jeong-Ho;Lee, Chun-Hee
    • Korean Journal of Environmental Agriculture
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    • v.12 no.2
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    • pp.144-152
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    • 1993
  • A bacterium which degrades efficiently synthetic detergents was isolated from the polluted waters, activated sludge of wastewater treatment plants or polluted soil. This bacterium showed considerably higher growth rate in the agar plate containing $2,000{\mu}g/ml$ of synthetic detergents than any other isolated strains, was identified as a Pseudomonas fluorescens or strains similar to it. The strain was named as a Pseudomonas fluorescens S1. Optimum pH and temperature for the growth of the Pseudomonas fluorescens S1 were pH 7.0 and $30^{\circ}C$, respectively. The strain was resistant to streptomycin and gentamycin, but sensitive to kanamycin. The strain was greatly resistant to zinc chloride, lead nitrate and copper sulfate, but unable to grow in the presence of relatively low concentrations of mercury chloride and silver nitrate. This strain utilized benzene, catechol, cyclohexane and xylene as a sole carbon source. The strain was well grown in the medium containing ABS 10,000${\mu}g$/ml. Degradation of ABS was 55% and 60% at 20${\mu}g$/ml and 100${\mu}g$/ml of ABS, respectively. Benzene ring was degraded 45% in 100${\mu}g$/ml of ABS. During the incubation of the strain in the medium containing ABS 100${\mu}g$/ml and COD 10,000${\mu}g$/ml for 4 days, degradation of ABS and COD were reduced to 40${\mu}g$/ml and 3,200${\mu}g$/ml, respectively. Total amino acid content of the Pseudomonas fluorescens S1 grown with 1,000${\mu}g$/ml of ABS was 115mg/g cell, whereas its content was decreased in the bacterium grown without synthetic detergent by 9.4%.

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Studies on Distribution, Characterization and Detoxification of Shellfish in Korea 1. A Study on the Distribution of Paralytic Shellfish Poison (한국산 주요패류에 대한 독의 분포, 특성 및 제독에 관한 연구 1. 마비성패류독의 분포에 관하여)

  • DONG Dong-Suck;SHIN Il-Shik;CHO Hak-Rae;KIM Ji-Hoe;PYEUN Jae-Hyeung;PARK Young-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.21 no.2
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    • pp.113-126
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    • 1988
  • Paralytic shellfish poison(PSP) accmulate in shellfish as a result of feeding toxic dinoflagellates. The shellfish do not seem to be harmed by the toxins, but become toxic to humans and other animals that feed on them. The purpose of this study was to investigate the distribution and changes of PSP by species of shellfish, collected area and collected month. Also, the correlation between PSP and toxic dinoflagellate, Protogonyaulax tamarensis, was investigated. Five hundred and six samples of 13 kinds of shellfish for PSP bioassay were collected at the shellfish growing area of Pusan, Masan, Chungmu, $Samch\check{o}npo, Y\check{o}su, Mokpo and Daech\check{o}n$ located in South Korea during the study period from May, 1985 to Octcber, 1987. Most of the samples submitted were free from PSP except sea mussel, short - necked clam and ark shell. Among the intoxicated samples, PSP was most often detected in sea mussel. PSP was detected mainly in spring$(February\~May)$ in the southern coast of Korea. In case of Pusan, exceptionally, toxic sea mussel have been found even June and July in 1987. The toxicity score of toxic shellfishes examined was ranged from 23.44 to $150.26{\mu}g/100g$ of edible meat and toxicity of sea mussel was higher than other toxic shellfishes. By the study of anatomical distribution of PSP in sea mussel collected at Masan in Febuary and March, 1986, the toxin accumulated in digestive gland was about $70\%$ of all. There was no significant correlation between toxicity of sea mussel and cell numbers of P. tamarensis that one of the causitive organism of PSP during the studying period in Masan area. There was almost no difference in toxicity of sea mussel by water depth of collection, but toxicity of surface shellfish was a little higher than those of 3.5, and 7.0m depth.

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Isolation and characterization of sigH from Corynebacterium glutamicum (Corynebacterium glutamicum의 sigH 유전자의 분리 및 기능분석)

  • Kim Tae-Hyun;Kim Hyung-Joon;Park Joon-Sung;Kim Younhee;Lee Heung-Shick
    • Korean Journal of Microbiology
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    • v.41 no.2
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    • pp.99-104
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    • 2005
  • Corynebacterial clones which exert regulatory effects on the expression of the glyoxylate bypass genes were isolated using a reporter plasmid carrying the enteric lacZ fused to the aceB promoter of Corynebacterium glutamicum. Some clones carried common fragments as turned out by DNA mapping technique. Subcloning analysis followed by the measurement of $\beta-galactosidase$ activity in Escherichia coli identified the region responsible for the aceB-repressing activity. Sequence analysis of the DNA fragment identified two independent ORFs of ORF1 and ORF2. Among them, ORF2 was turned out to be responsible for the aceB-repressing activity. ORF1 encoded a 23,216 Da protein composed of 206 amino acids. Sequence similarity search indicated that the ORF may encode a ECF-type $\sigma$ factor and designated sigH. To identify the function of sigH, C. glutamicum sigH mutant was constructed by gene disruption technique and the sigH mutant showed growth retardation as compared to the wild type strain. In addition, the mutant strain showed sensitivity to oxidative-stress generating agent plumbagin. This result imply that sigH is probably involved in the stress response occurring during normal cell growth.

Identification of Novel Psychrotolerant Bacterial Strain and Production of $\beta-Galactosidase$ (새로운 저온 내성세균의 동정과 $\beta-Galactosidase$ 생산)

  • Park, Jeong-Woon;Yoo, Jae-Soo;Roh, Dong-Hyun
    • Korean Journal of Microbiology
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    • v.42 no.1
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    • pp.40-46
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    • 2006
  • Galactose joined to glucose by a $\beta(1\rightarrow4)$ glycosidic bond makes lactose and this disaccharide is rich in milk. It is known that lacotse is hydrolyzed to each monomeric sugar by either lactase in human or $\beta-galactosidase$ in bacteria. Ingestion of milk by lactase-deficient persons causes a temporary diarrhea and subsequent chronic diarrhea results in colitis with chronic inflammation. We isolated a $\beta-galactosidase$ producing psycrotolerant strain AS-20 from near cattle shed and investigated the growth at various temperature conditions. Whereas Escherichia coli strains did not grow at $10^{\circ}C$, the AS-20 strain could grow well at this low temperature and showed optimal growth at $30^{\circ}C$. The isolated strain was identified as 97% Hafnia alvei by biochemical properties. This strain could ferment glucose, lacotse, maltose, mannitol, xylose, ONPG, rhamanose and L-arabinose, and decarboxylate lysin and ornithine. To confirm the identity of isolated strain we amplified 16S rDNA by PCR and searched similarity of the 1426 bp DNA sequcence with Genbank database. The strain AS-20 showed 99% similarity with Hafnia alvei. The activity of $\beta-galactosidase$ was 1.5 times higher when the cell was grown at 10 or $20^{\circ}C$ than at $30^{\circ}C$. The highest enzyme activity of AS-20 was also much higher than that of E. coli, which was grown at $30^{\circ}C$.