• Title/Summary/Keyword: biochemical composition

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Fermentation Process Development of Recombinant Hansenula polymorpha for Gamma-Linolenic Acid Production

  • Khongto, B.;Laoteng, K.;Tongta, A.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.11
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    • pp.1555-1562
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    • 2010
  • Development of the strain and the fermentation process of Hansenula polymorpha was implemented for the production of ${\gamma}$-linolenic acid ($GLA,\;C18:3{\Delta}^{6,9,12}$), an n-6 polyunsaturated fatty acid (PUFA) that has been reported to possess a number of health benefits. The mutated ${\Delta}^6$-desaturase (S213A) gene of Mucor rouxii was expressed in H. polymorpha under the control of the methanol oxidase (MOX) promoter. Without the utilization of methanol, a high-cell-density culture of the yeast recombinant carrying the ${\Delta}^6$-desaturase gene was then achieved by fed-batch fermentation under glycerol-limited conditions. As a result, high levels of the ${\Delta}^6$-desaturated products, octadecadienoic acid ($C18:2{\Delta}^{6,9}$), GLA, and stearidonic acid ($C18:4{\Delta}^{6,9,12,15}$), were accumulated under the derepression conditions. The GLA production was also optimized by adjusting the specific growth rate. The results show that the specific growth rate affected both the lipid content and the fatty acid composition of the GLA-producing recombinant. Among the various specific growth rates tested, the highest GLA concentration of 697 mg/l was obtained in the culture with a specific growth rate of 0.08 /h. Interestingly, the fatty acid profile of the yeast recombinant bearing the Mucor ${\Delta}^6$-desaturase gene was similar to that of blackcurrant oil, with both containing similar proportions of n-3 and n-6 essential fatty acids.

Biochemical Properties of Starch Granule Non-Digestive Enzyme(SGNA) of Bacillus polymyxa No.26

  • Sohn, Cheon-Bae;Kim, Myung-Hee;Bae, Jung-Surl
    • Journal of Microbiology and Biotechnology
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    • v.2 no.3
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    • pp.189-196
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    • 1992
  • A $\alpha$-l, 4-D-glucan maltohydrolase $(\beta$-amylase), secreted by the mesophilic aerobic bacterium Bacillus polymyxa No.26, was purified and characterized. The enzyme production was increased after a logarithmic phase of bacterial growth and paralleled with the onset of bacterial sporulation. By applying anion exchange chromatography and gel filtration the enzyme was purified 16.7-fold and had a specific activity of 285.7 units/mg. Two enzyme activities were eluted on a column of DEAE-Sephadex chromatography, and they were designated as E-I for a major enzyme peak and E-II for a minor peak. Of them, E-I enzyme peak was further purified by using gel chromatography. The molecular mass of this enzyme was determined to be 64, 000 daltons and consisted of a single subunit, showing an isoelectric point of 8.9. The enzyme was able to attack specifically the $\alpha$-l, 4-glycosidic linkages in soluble starch and caused its complete hydrolysis to maltose and $\beta$-limited dextrin. This amylolytic enzyme displayed a temperature optimum at $45^\circ{C}$ and a pH optimum at 7.0. The amino acid composition of the purified enzyme was quite similar to the other bacterial $\beta$-amylases reported. Surprisingly, the purified enzyme from this aerobe only exhibited hydrolytic activity on soluble starch, not on starch granules. The degradation of from starch by $\beta$-amylase was greatly stimulated by pullulanase addition. These results differentiated from other $\beta$-amylases reported. Based on a previous result that showed the enzyme system involves in effective degradation of raw starch granules, this result strongly suggested that the purified enzyme (E-I) can be a synergistic part of starch granule-digestion and E-II plays a crucial role in digestion of starch granules.

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Preparation and Characteristics of Inorganic Pigments (무기안료의 제조 및 특성에 관한 연구)

  • Oh, Sae-Joong
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.14 no.6
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    • pp.3102-3107
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    • 2013
  • Inorganic pigments were prepared from $Al_2O_3$, CoO, $Cr_2O_3$, ZnO, $SrO_2$ and Kaolin. The pigment was prepared through the processes of mixing and crushing of the raw materials, sintering, milling and drying. The color of the pigments was changed from blue to green as $Cr_2O_3$ contents in the pigment increased. The color became brighter irrespective of the pigment composition as pigment particle size became smaller. Maximum sintering temperature was the most suitable at around $1,250^{\circ}C$. In the test of compatability between the pigment and glaze, the color became close to green as $Cr_2O_3$ contents in the pigment increased, and became close to blue as CoO contents increased.

Effect of LED Lighting Intensity on Productivity, Blood Parameters and Immune Responses in Dairy Cows (LED 점등 세기가 젖소의 생산성, 혈액 매개변수 및 면역 반응에 미치는 영향)

  • Park, Jin-Ryong;Belal, Shah-Ahmed;Na, Chong-Sam;Shim, Kwan-Seob
    • Korean Journal of Organic Agriculture
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    • v.27 no.2
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    • pp.161-171
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    • 2019
  • A light-emitting diode (LED) is most efficient in terms of economic benefits. The aim of the present study was to evaluate the effects of LED lighting intensity on milk production, milk composition, and the immune response of Holstein cows. Thirty-eight lactating cows were assigned to four experimental groups: control group (exposed natural daylight without LED light) and three LED treatment group (50 lux; natural daylight and 6 h LED light of 50 lux intensity, 100 lux; natural daylight and 6 h LED light of 100 lux intensity, 200 lux; natural daylight and 6 h LED light of 200 lux intensity). There was a significant effect on the decrease ratio in milk production in 200 lux group. Lactose was significantly increased in 100 lux than the other LED treated groups. In addition to the hemolytic biochemical analysis, AST and β-HB were significantly increased in the 50 lux than the other groups. Prolactin and IGF-1 levels were significantly increased in 200 lux compared to other groups. Besides, cortisol was significantly higher in 50 lux and lowered in the 200 lux than the control, while IgA and IgG were not significant among the groups. Therefore, it was concluded that LED lighting intensity had some impact on blood parameters and immune responses in dairy cows with increased milk production.

Characteristics of a New Obligate Methanol-Oxidizing Bacterium (새로운 절대 메탄올 산화세균의 분리 및 특성)

  • Kim, Si-Wook;Park, Yong-Ha
    • Korean Journal of Microbiology
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    • v.31 no.4
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    • pp.261-266
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    • 1993
  • A new methyltrophic bacterium which utilizes methanol as a sole source of carbon and energy was isolated from soil. It was Gram-negative, nonmotile, nonspore-forming rod, and strictly aerobic bacterium. Catalase and oxidase activities were present. Nitrate was reduced to nitrite. Vitamins and other growth factors were not required. Generation time was 1.6 hr under the optimal condition. The isolate assimilated methanol via the ribulose mono-phosphate pathway (Enter-Doudoroff varient) and did not have .alpha.-ketoglutarate dehydrogenase. It assimilated ammonia through glutamate dehydrogenase. The guanine plus cytosine content of the DNA was 61.0 mol%. The celular fatty acid composition was primarily straight-chain saturated $C^{16 : 0}$ acids (palmitic acids) and unsaturated $C_{16 :1}$ acid (palmitoleic acids), and the isolate also contained two unidentified $C_{17}$ branched fatty acids. The major ubiquinone was Q-8, and Q-6 and Q-7 were present as minor components. Phosphatidylethanolamine and phosphatidylglycerol were predominantly present, and diphosphatidyglycerol was also detected. Based on the physiological and biochemical properties, the isolate was assigned to a novel species of the genus Methylobacillus, Methylobacillus methanolovorus sp. nov.

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Isolation and Identification of ${\gamma}$-Cyclodextrin Glycosyltransferase-Producing Bacteria, and Its Production Conditions (${\gamma}-Cyclodextrin$ Glycosyltransferase 생산균주의 분리, 동정 및 효소 생산조건)

  • Kim, Myung-Hee;Lim, Young-Hee;Bae, Kyung-Sook;Oh, Tae-Kwang;Sohn, Cheon-Bae
    • Applied Biological Chemistry
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    • v.40 no.6
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    • pp.459-464
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    • 1997
  • A cyclodextrin glycosyltransferase-producing bacterium was newly isolated from soil using alkaline pH medium containing 1% $Na_2CO_3$. The isolated strain was identified as Bacillus brevis by morphological and biochemical characteristics, and fatty acid composition and designated Bacillus brevis CD162. The strain showed the best enzyme production of 0.9 unit/ml after 96 hrs of culture at $30^{\circ}C$ in a medium of 2.0% soluble starch, 0.75% yeast extract, 0.5% bacto peptone, 0.2% $K_2HPO_4$ 0.05% $MgSO_4{\cdot}7H_2O$, and 1.5% $Na_2CO_3$ at initial pH 10.2.

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Carrot Juice Administration Decreases Liver Stearoyl-CoA Desaturase 1 and Improves Docosahexaenoic Acid Levels, but Not Steatosis in High Fructose Diet-Fed Weanling Wistar Rats

  • Mahesh, Malleswarapu;Bharathi, Munugala;Reddy, Mooli Raja Gopal;Kumar, Manchiryala Sravan;Putcha, Uday Kumar;Vajreswari, Ayyalasomayajula;Jeyakumar, Shanmugam M.
    • Preventive Nutrition and Food Science
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    • v.21 no.3
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    • pp.171-180
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    • 2016
  • Non-alcoholic fatty liver disease (NAFLD) is one of the most prevalent liver diseases associated with an altered lifestyle, besides genetic factors. The control and management of NAFLD mostly depend on lifestyle modifications, due to the lack of a specific therapeutic approach. In this context, we assessed the effect of carrot juice on the development of high fructose-induced hepatic steatosis. For this purpose, male weanling Wistar rats were divided into 4 groups, fed either a control (Con) or high fructose (HFr) diet of AIN93G composition, with or without carrot juice (CJ) for 8 weeks. At the end of the experimental period, plasma biochemical markers, such as triglycerides, alanine aminotransferase, and ${\beta}$-hydroxy butyrate levels were comparable among the 4 groups. Although, the liver injury marker, aspartate aminotransferase, levels in plasma showed a reduction, hepatic triglycerides levels were not significantly reduced by carrot juice ingestion in the HFr diet-fed rats (HFr-CJ). On the other hand, the key triglyceride synthesis pathway enzyme, hepatic stearoyl-CoA desaturase 1 (SCD1), expression at mRNA level was augmented by carrot juice ingestion, while their protein levels showed a significant reduction, which corroborated with decreased monounsaturated fatty acids (MUFA), particularly palmitoleic (C16:1) and oleic (C18:1) acids. Notably, it also improved the long chain n-3 polyunsaturated fatty acid, docosahexaenoic acid (DHA; C22:6) content of the liver in HFr-CJ. In conclusion, carrot juice ingestion decreased the SCD1-mediated production of MUFA and improved DHA levels in liver, under high fructose diet-fed conditions. However, these changes did not significantly lower the hepatic triglyceride levels.

Clinical Effect of Korean Red Ginseng on Osteoporosis (홍삼의 골다공증에 대한 임상적 효능 연구)

  • 김남현;이환모
    • Journal of Ginseng Research
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    • v.22 no.2
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    • pp.114-121
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    • 1998
  • Osteoporosis is defined as a pathologic condition in which there occurs no change in the chemical composition of the bone, while bone resorption is abnormally increased. This compares with osteogenesis, which leads to a decrease in the amount of bone. Though many varieties of therapies have been tried, no fully effective method has been found. Korean red ginseng is an important variety of Korean ginsengs and many studies have been performed to investigate its effectiveness. But there has been no report on red ginseng's effects on osteoporosis. The purpose of this study was to determine the clinical effects of Korean red ginseng on postmenopausal osteoporosis patients. The study was designed as a double-blind study. Group I consisted of 30 postmenopausal osteoporosis patients who were administered red ginseng extracts and 15 patients who were administered maltose capsules, were included in group II. The following results were obtained after one-year administration to both groups. 1. There were no significant differences in the degree of wedging, number of compression fractures in the vertebral body, trabeculations, and Cobb's angles between the experimental and control groups. 2. Even though bone mineral density increased in the lumbar spine, and decreased in the femur, there was no significant statistical difference. 3. The serum levels of parathyroid hormone (PTH), calcium, phosphate, and alkaline phosphates showed no difference between the two groups regardless of treatment. 4. Urine Deoxy-pyridinoline (DPYD) decreased in the red ginseng group while it increased in the maltose group, but statistically there was no significant difference between the two groups. 5. The clinical presentations showed no significant differences using Visual Analog Scale. 6. The specific complications related with the long-term use of red ginseng did not occur in all patients. In conclusion, there were no statistically significant differences in clinical presentations, biochemical, and radiological studies between the red gins eng and maltose groups. No definite effectiveness of red ginseng on patients with osteoporosis was found.

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Comparative Biochemical Properties of Proteinases from the Hepatopancreas of Shrimp. -I. Purification of Protease from the Hepatopancreas of Penaeus japonicus-

  • Choi Sung-Mi;Oh Eun-Sil;Kim Doo-Sang;Pyeun Jae-Hyeung;Cho Deuk-Moon;Ahn Chang-Bum;Kim Hyeung-Rak
    • Fisheries and Aquatic Sciences
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    • v.1 no.2
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    • pp.201-208
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    • 1998
  • A protease, which had no tryptic and chymotryptic activity, was purified from the hepatopancreas of shrimp, P. japonicus, through ammonium sulfate fractionation, Q­Sepharose ionic exchange, benzamidine Sepharose 6B affinity, and Sephacryl S-100 gel chromatography. Molecular weight (M.W.) of the protease was estimated to be 24 kDa by gel filtration and showed a single peptide band by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). The protease had a low ratio of acidic to basic amino acids, which is different with pro teases from marine animals. The enzyme was partially inhibited by benzamidine, tosyl-L-lysine chioromethyl ketone (TLCK), phenylmethylsulfonyl fluoride (PMSF), soybean trypsin inhibitor (SBTI), and pepstatin. The enzyme did not have any activity against benzoyl-D,L-arginine p-nitroanilide (BAPNA) or benzoyl-L-tyrosine ethyl ester (BTEE) which is a specific substrate of trypsin and chymotrypsin, respectively. However, the enzyme showed activity forward N-CBZ-L-tyrosine p-nitrophenyl ester (CBZ-Tyr-pNE), N­CBZ-L-tryptophan p-nitrophenyl ester (CBZ-Trp-pNE), and N-CBZ-L-proline p-nitrophenyl ester (CBZ-Pro-pNE). The protease did not showed tryptic and chymotryptic activity, which was not reported in shrimp hepatopancreas.

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Molecular Characterization of the Levansucrase Gene from Pseudomonas aurantiaca S-4380 and Its Expression in Escherichia coli

  • Jang, Eun-Kyung;Jang, Ki-Hyo;Isaac Koh;Kim, In-Hwan;Kim, Seung-Hwan;Kang, Soon-Ah;Kim, Chul-Ho;Ha, Sang-Do;Rhee, Sang-Ki
    • Journal of Microbiology and Biotechnology
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    • v.12 no.4
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    • pp.603-609
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    • 2002
  • DFA IV is di-D-fructose-2,6':6,2'-dianhydride, consisting of two fructose residues. It can be enzymatically synthesized from levan by levan fructotransferase, and can be used for mineral absorption. Understanding of the structure and composition of levan is important to obtain high-level production of DFA IV. A bacterial strain, Pseudomonas aurantiaca 5-4380, was identified to produce low-branched levan, and the levansucrase gene (lsch) from this bacterium was found to be composed of 1,275 Up coding for a protein of 424 amino acids, with an estimated molecular weight of 47 kDa. The bacterial levansucrase gene was expressed in Escherichia coli DH5${\alpha}$ by its own promoter and lac promoter. The recombinant levansucrase was produced in soluble form with 170U of levansucrase activity from 1-ml E. coii culture broth. The expressed enzyme from the clone showed similar biochemical properties, such as size of active levansucrase, degree of branching, and optimum temperature, with P.aurantiaca 5-4380 levansucrase.