• Title/Summary/Keyword: biochemical properties

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Description and Application of a Marine Microalga Auxenochlorella protothecoides Isolated from Ulleung-do (울릉도 거북바위 조수웅덩이에서 분리된 해양 미세조류 옥세노클로렐라 프로토테코이드 균주의 기술 및 응용)

  • Jang, Hyeong Seok;Kang, Nam Seon;Kim, Kyeong Mi;Jeon, Byung Hee;Park, Joon Sang;Hong, Ji Won
    • Journal of Life Science
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    • v.27 no.10
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    • pp.1152-1160
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    • 2017
  • A unicellular green alga was axenically isolated from a tidal pool on Ulleung-do, Korea. Morphological, molecular, and biochemical analyses revealed that the isolate belonged to Auxenochlorella protothecoides. The current study is the first record of this species in Korea. The microalgal strain was named as A. protothecoides MM0011 and its growth, lipid and pigment compositions, and biomass properties were investigated. The strain is able to thrive in a wide range of temperatures ($5{\sim}35^{\circ}C$) and to withstand up to 1.5 M NaCl. The results of GC/MS analysis showed that the isolate was rich in nutritionally important polyunsaturated fatty acids (PUFAs). Its major fatty acids were linoleic acid (27.6%) and ${\alpha}-linolenic$ acid (39.6%). Thus, this indigenous microalga has potential as an alternative source of ${\omega}3$ and ${\omega}6$ PUFAs, which currently come from fish and plant oils. Also, the HPLC analysis revealed that the value-added antioxidant, lutein, was biosynthesized as the accessory pigments by the microalga. A proximate analysis showed that the volatile matter content was 85.6% and an ultimate analysis indicated that the gross calorific value was $20.3MJ\;kg^{-1}$. Since 40.5% of total nitrogen and 27.9% of total phosphorus were removed from the medium, respectively, it also has potential as a feedstock for biofuel applications which could be coupled to wastewater treatment. In addition, the biomass may also serve as an excellent animal feed because of its high protein content (51.4%). Therefore, A. protothecoides MM0011 shows promise for application in production of microalgae-based biochemicals and as a biomass feedstock.

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.

Mineral Composition and Physical Properties Change of Chicken Muscle (계육(鷄肉)의 무기물(無機物) 조성(組成)과 물리적(物理的) 성질(性質)의 변화(變化))

  • Chung, Byoung-Yeol;Yang, Cheul-Young
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.11 no.4
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    • pp.47-52
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    • 1982
  • This experiment was carried out to investigate the minerals content and post-mortem biochemical change of broiler leg and breast muscle during hot and cold storage period. The leg and breast muscle samples were stored at $2^{\circ}C$and $30^{\circ}C$ for 32 hours, and analyzed 0, 4, 8, 32 hours after slaughter, the results obtained as follows (1) Minerals composition were different between breast and leg muscle. (2) Total nitrogen content changed according to the storage period and temperature in the breast (P<0.005) and the leg (P<0.005) (3) The Content of phosphate (P<0.005), magnesium (P<0.005), sodium (P<0.025) in breast muscle was higher than in the leg mucle, and did not change according to the storage period and temperature. (4) Calcium and iron contents were not different between breast and leg muscle but significantly changed according to the storage period and temperature. (5) Sodium content was not different among the various factors, and pH was different between leg and breast (P<0.005) muscle, and ultimate pH was appeared at 8 hours of slaughter, but 20 hours of slaughter in hot stored breast muscle. (6) pH decline in cold stored breast muscle was faster than in the hot stored. (7) Penetration value was higher in the breast muscle than in the leg muscle (P<0.005) (8) Regormortis were not occurred in cold stored breast muscle and hot stored leg muscle, but occurred in cold stored leg muscle and hot stored breast muscle at 4 hours after slaughter.

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Effect of Green Tea Extract on Cisplatin- or Doxorubicin-Induced Cytotoxicity in Human Lung Cancer Cell Lines (사람 폐암 세포주에서 시스플라틴이나 독소루비신의 세포독성에 미치는 녹차 추출물의 영향)

  • Lee, Byoung-Rai;Park, Jae-Yoon;Park, Pyoung-Sim
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.5
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    • pp.619-624
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    • 2011
  • Tea extract (TE) has been shown to have anti-tumor properties in a wide variety of experimental systems. We evaluated green tea extract (GTE) as a biochemical modulator for the antitumor activity of cisplatin and doxorubicin in the treatment of human lung cancer A549 cells. Cells were grown in RPMI-1640 medium supplemented with 10% (v/v) heat-inactivated fetal bovine serum and two antibiotics (100 units/mL penicillin and $100\;{\mu}g$/mL streptomycin). Two types of TE, epigallocatechin galate (EGCG) and GTE, were used in this experiment. The cells were seeded at $1{\times}10^4$ cells/well in the RPMI-1640 media with or without TE ($100\;{\mu}g$/mL) and then treated with different concentrations of doxorubicin ($0{\sim}14\;{\mu}g$/mL) or cisplatin ($0{\sim}35\;{\mu}g$/mL). After incubation in 5% $CO_2$ at $37^{\circ}C$ for 24 hr, cell viability was determined with a MTT assay. We used a Western blot to detect the influence of EGCG and GTE on the expression of p53 and caspase-3 genes in the A549 cells. A549 cell viability decreased to 15% with a $10\;{\mu}g$/mL concentration of cisplatin, and to 21% with a $8\;{\mu}g$/mL concentration of doxorubicin, as measured with the MTT assay. However, pre-treatment of the cells with EGCG ($100\;{\mu}g$/mL) or GTE ($100\;{\mu}g$/mL) resulted in decreased cell viability with $6\;{\mu}g$/mL of cisplatin and $4\;{\mu}g$/mL of doxorubicin. There was no apparent change in cell viability between EGCG or GTE administration in cisplatin- or doxorubicin-induced cytotoxicity in A549 cells. The levels of p53 and caspase-3 in the A549 cells increased with both EGCG and GTE treatment. We found that GTE could potentially affect cisplatin- or doxorubicin-induced cytotoxicity of A549 cells, which may be useful in the chemotreatment of cancer.

Properties of Blood Pressure and Routine Laboratory Test Results by the Status of Smoking and Alcohol Intakes in Male Workers (일부 산업장 남성근로자들의 흡연,음주실태에 따른 혈압 및 혈액검사치의 특성)

  • Yoo, Chang-Kyun;Jeong, Yong-Jun
    • Korean Journal of Health Education and Promotion
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    • v.20 no.1
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    • pp.131-145
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    • 2003
  • This study was conducted to estimate the incidence and the degree of cigarette smoking and drinking among working men, and then to investigate the effects on blood pressure, various hematological indices and blood chemistry. The sample consisted of 2,287 male workers who had undertaken a general health check-up during the two-year period from January, 2000 to December, 2001. Such factors as blood pressure, blood glucose, lipid profiles, and liver function tests were determined and then analyzed with respect to the subjects smoking and/or drinking status. The major findings from this study are: 1. The drinking and smoking status have shown that 52.7% of participants were in the habit of both drinking and smoking while 11.6% were not associated with either. On the other hand 25.4% were involved only in drinking and 10.2% only in smoking. In the group smoking over 21 cigarettes per day over 30, the age group occupied the largest proportion at 20.1%. 2. Regarding the relationship between smoking and/or drinking status, and blood pressure, hematology and blood chemistry, the smoking and/or drinking group had significantly higher levels of blood pressure, both systolic and diastolic, Hb & Hct, TG, LDL-C, SGOT, and ${\gamma}$-GTP, than the non-smoking and/or non-drinking group. But HDL-C was significantly lower in the smoking group and significantly higher in the drinking group than the non-smoking/non-drinking group. 3. Regarding amount smoked, a larger number of cigarettes per day was significantly associated with the higher levels of blood pressure, systolic and diatolic, TG, TC, LDL-C, Hb, Hct, and ${\gamma}$-GTP. As for the amount druck, an increasing amount of alcohol intake was associated with rising levels of blood pressure, systolic and diatolic, TG, TC, LDL-C, HDL-C, Hb, SGOT, and ${\gamma}$-GTP. 4. Regarding the correlation among all the variables stated above, the smoking and drinking amount was shown to be in the positive correlation with blood pressure, both systolic and diastolic, TG, TC, Hb, and ${\gamma}$-GTP. On the contrary, LDL-C and HDL-C was in a positive correlation only with the amount drunk amount, and Hct only with the amount smoked. 5. As with systolic and diastolic blood pressure, the odds ratio of the smoking group was 2.35 and 2.58 compared to the non-smoking/drinking group. whereas it was 1.47 and 1.75 in the smoking/drinking group. Concerning serum lipids, the smoking/drinking group had 1.97 times the levels of TG in the non-smoking/non-drinking group, though the smoking group had 1.55 times the levels of HDL-C in the non-smoking/non-drinking group. As with liver function test results, the drinking group had 2.50 times and the smoking/drinking group had 4.41 times the levels of ${\gamma}$-GTP in the non-smoking/drinking group. respectively. The above results revealed that smoking and alcohol intake were effected the results of blood pressure and laboratory tests. Specifically, not only the smoking/drinking group but also those only smoking or only drinking were not as desirable as the non-smoking and non-drinking group to the results of blood pressure and laboratory tests.

Selectivity of Bleaching Herbicides Caused by Physiological Differences between Rice and Barnyardgrass (벼와 피의 생리적(生理的) 차이(差異)에 의한 백화형(白化型) 제초제(除草劑)의 선택성(選擇性))

  • Na, J.Y.;Kim, J.S.;Kim, T.J.;Cho, K.Y.;Pyon, J.Y.
    • Korean Journal of Weed Science
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    • v.12 no.2
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    • pp.89-101
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    • 1992
  • In this study, various physiological and biochemical experiments were conducted to know whether the selectivity between rice and barnyardgrass treated with bleaching herbicides containing diphenyl ether compounds was also partly based on their basic physiological proterties such as peroxidation ability, membrane stability or antioxidant system. It seemed to be partly based on the differences of their physiological characteristics that barnyardgrass was commonly more susceptible to most of the bleaching herbicides than rice. The scenescence of intact leaf segment was more rapid in barnyardgrass than in rice, indicating that barnyardgrass is weak at early stage. Also pigment metabolic ability, antioxidant enzyme activities(peroxidase, catalase, superoxide dismutase, glutathione reductase) and antioxidant content (tocopherol, ascorbic acid, glutathione, carotenoids) were lower in barnyardgrass on the basic of fresh weight. However, lipoxygenase activity and the content of unsaturated fatty acid which is vulnerable to oxygen radicals were higher in barnyardgrass, suggesting that barnyardgrass seedling bave a properties easy to be peroxidized. The differences of PPIX (protoporphyrin IX) or carotenoid content, which are the primary substances inducing herbicide activity, were not related to the selectivity between rice and barnyardgrass.

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Characterization of a Novel Fibrinolytic Enzyme Produced from Bacillus subtilis BK-17 (Bacillus subtilis BK-17 유래 혈전용해 효소의 특성)

  • Hyun Bek;Lim Hak-Seob;Chung Kyung Kae;Choi Yung Hyun;Choi Byung Tae;Seo Min-Jeong;Kim Ji-Eun;Ryu Eun-Ju;Huh Man Kyu;Joo Woo Hong;Jeong Young Kee
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.987-993
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    • 2005
  • A bacterium, producing a fibrinolytic enzyme, was screened from a decaying rice plant. The bacterium was identified as Bacillus subtilis by morphological, biochemical, and physiological properties and named Bacillus subtilis BK-17. The fibrinolytic enzyme (BK) was purified from supernatant of Bacillus subtilis BK-17 culture broth. The molecular weight was 31 kDa as determined by SDS-PAGE. The effect of temperature, pH, and plasminogen on the activity of the bacillokinase (BK) was analysed and the activity was compared with urokinase. The optimal temperature and pH were $50^{circ}C$ and pH 7, pH 8, respectively. The BK activity was inhibited to $45\%$, $35\%$, and $23\%$ with 1mM EDTA, $Zn^{2+}$, and $Ca^{2+}$, respectively. However, $Mg^{2+}$, $Mn^{2+}$, and $Co^{2+}$ ions did not have any significant effect on the enzyme activity The BK showed the artivity in the both plates, plasminogen-free fibrin plate and plasminogen-rich fibrin plate. The result indicates that the BK can directly act the fibrin. In comparison of fibrinolytic activity with urokinase on the fibrin plate, the BK shows about 20 folds higher activity than that of the urokinase.

A Report on the Shigella Cultures Isolated in Korea (1972) (1972년(年) 한국(韓國)에서 분리(分離)된 이질균(痢疾菌)에 관(關)한 보고(報告))

  • Ryu, Young-Hat;Kim, Soon-Hee
    • The Journal of the Korean Society for Microbiology
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    • v.8 no.1
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    • pp.7-11
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    • 1973
  • The authors identified eighty-eight Shigella cultures among about four thousands specimens collected from all over the country in 1972. Of eighty-eight cultures, seventy-seven cultures belonged to Shigella flexneri and eleven cultures to Shigella sonnei. None of cultures belonging to either subgroup A or C was detected in 1972. Of seventy-seven cultures of Shigella flexneri one was $B_{1b}$, fifty-six were $B_{2a}$, nine were $B_{3a}$, six were $B_{4a}$, three were By and one was each of $B_{3b}$ and $B_{3c}$. Of eleven cultures of Shigella sonnei seven cultures appeared to be phase I and the others phase II. Although there was quite a difference found in the incidence of isolating Shigella organisms between different areas as shown in Table 1, it would not be possible to understand that there might not have been the cases or carriers of Shigella in the areas where the organisms were not isolated in 1972. Concerning the biochemical properties it was not possible to compare the results obtained from the decarboxylase and dihydrolase tests with them obtained in previous years except that of lysine decarboxylase tests since they were not reported individually by the different serotypes in the previous reports. These results obtained in 1972 would be the data for the future comparison. In regards to the antibiotics-sensitivity of Shigella cultures the most of them showed sensitive results to nitrofurantoin, ampicillin, cephalosporin, gentamycin and geopen, and the majority of them appeared to be resistant to cloxacillin, tetracycline and streptomycin by means of the In Vitro tests.

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Plant Growth Promoting Effect and Antifungal Activity of Bacillus subtilis S37-2 (Bacillus subtilis S37-2 균주의 항진균활성 및 식물생육촉진 효과)

  • Kwon, Jang-Sik;Weon, Hang-Yeon;Suh, Jang-Sun;Kim, Wan-Gyu;Jang, Kab-Yeul;Noh, Hyung-Jun
    • Korean Journal of Soil Science and Fertilizer
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    • v.40 no.6
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    • pp.447-453
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    • 2007
  • With a broad objective for the development of microbial based fertilizers, a total of 373 strains were isolated from rhizoplane and rhizosphere of pepper, tomato, lettuce, pasture, and grass. The efficacy of the isolates to augument overall plant growth was evaluated. After screening for their plant growth promotion and antagonistic properties in vitro efficient strains were further selected. The most efficient strains was characterized by 16S rRNA gene sequences and biochemical techniques and was designated as Bacillus subtilis S37-2. The strains facilitated plant growth and inhibited the plant phathogenic fungi such as Fusarium oxysporum (KACC 40037, Rhizoctonia solani (KACC 40140), and Sclerotinia sclerotiorum (KACC 40457). Pot based bioassay using lettuce as test plant was conducted by inoculating suspension ($10^5$ to $10^8cells\;mL^{-1}$) of B. subtilis S37-2 to the rhizosphere of lettuce cultivated in soil pots. Compared with non-inoculated pots, marked increase in leaf (42.3%) and root mass (48.7%) was observed in the inoculation group where the 50ml of cell mixture ($8.7{\times}10^8cells\;ml^{-1}$) was applied to the rhizosphere of letuce either once or twice. Antagonistic effects of B. subtilis S37-2 strain on S. sclerotiorum (KACC 40457) were tested. All the tested lettuce plants perished after 9 days in treatment containing only S. sclerotiorum, but only 17% of lettuce was perished in the inoculation plot. B. subtilis grew well in the TSB culture medium. The isolates grew better in yeast extracts than peptone and tryptone as nitrogen source. The growth rate was 2~4 times greater at $37^{\circ}C$ as compared with $30^{\circ}C$ incubation temperature. B. subitlis S37-2 produced $0.1{\mu}g\;ml^{-1}$ of IAA (indole 3-acetic acid) in the TSB medium containing L-tryptophan($20mg\;L^{-1}$) in 24 hours.