• Title/Summary/Keyword: Protein engineering

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Enhanced Anaerobic Degradation of Food Waste by Employing Rumen Microorganisms (Rumen 미생물을 이용한 주방폐기물 혐기성소화의 효율증진 방안)

  • Shin, Hang-Sik;Song, Young-Chae;Son, Sung-Sub;Bae, Byung-Uk
    • Journal of the Korea Organic Resources Recycling Association
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    • v.1 no.1
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    • pp.103-113
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    • 1993
  • Every year, over $3.37{\times}10^7$ ton of municipal solid waste is generated in Korea, of which about 28% is organic food waste from restaurant, dining halls and households etc. Methane conversion of the food waste by anaerobic digestion could be a viable approach for energy recovery as well as safe disposal of the waste. However, as food waste is composed of highmolecular complex polymers such as cellulose, lignin and protein, anaerobic digestion of food waste has not been efficient in terms of volumetric loading rate, solid retention time and extent of anaerobic degradation. In this research, the improved anaerobic degradation of food waste was attemped by applying rumen microorganisms to anaerobic digestion. Acidification efficiency of food waste by rumen microorganisms was compared with that of conventional acidogenesis. And optimum acidification conditions by rumen microorganisms were also determined. For the experiments, anaerobic batch reactors of 600 mL was fed with the processed (dried and milled) food waste obtained from a restaurant. Ultimate volatile fatty acid (VFA) yield produced by rumen microorganisms was about 8.4 meq VFA/g volatile solid (VS) that is 95% of the theoretical value. This yield was not much different from that of conventional acidogenesis, but hydrolysis rate was about twice faster. Cumulative VFA concentration increased from 66 meq/L to 480 meq/L, when the initial TS was increased from 1% to 15%. But VFA yield at 15% TS was half of that at 1% TS. This inhibition on the acidification might be caused by the rapid drop of pH and higher concentration of nonionized VFA. Optimal pH and temperature range for the acidification were about 6.0~7.5 and $35{\sim}45^{\circ}C$, respectively.

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Development of medium for Pleurotus eryngii cultivation using Kojongsi persimmon peels (감과피를 이용한 큰느타리버섯 배지 개발)

  • Kim, Chul Hwan;Kim, Hye Soo;Kim, Hong Chul;Kwon, Hyun Sook;Cheong, Jong-Chun;Kong, Won-Sik;Cho, Soo Jeong
    • Journal of Mushroom
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    • v.13 no.4
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    • pp.314-318
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    • 2015
  • This study was carried out to investigated the avaiability of dried Koojongsi persimmon peels (KPP) as a useful mushroom medium using Pleurotus eryngii ASI 2312. Mushroom cultivation medium used in this study was mixed with medium mixture, corn cob and sawdust (220:65:15, v/v). Dried KPP was replaced mushroom cultivation medium (control) with 5, 10, 15, 20, 30, 40, 50% dried KPP. The T-N content of dried KPP treatments decreased to increase replaced ratio of the dried KPP and C/N ratio was increased to increase replaced ratio of the dried KPP. But T-C content of dried KPP treatments was similar to untreated control. The average cultivating periods of mycelium on dried KPP treatments was delayed to increase replaced ratio of the dried KPP and cultivating periods was delayed over 30% dried KPP treatments. The length of stipe of dried KPP treatments was longer than that of the untreated control to increase replaced ratio of the dried KPP and thickness of stipes was tend to be thinner than that of the untreated control to increase replaced ratio of the dried KPP. The moisture, carbohydrate, crude protein and crude ash content of mycelial were similar to untreated control, but crude fatty acid was increased to increase replaced ratio of the dried KPP. The ${\beta}-glucan$ content of 10% and 15% treatments were higher than untreatment control. The results based on cultivation yield and ${\beta}-glucan$ content indicated that optimal mixture ratio dried KPP was 15%.

Anti-inflammatory Effect of Lactuca sativa L. Extract in Human Umbilical Vein Endothelial Cells and Improvement of Lipid Levels in Mice Fed a High-fat Diet (상추 추출물(Lactuca sativa L.)의 혈관내피세포에서 항염증 작용과 고지방 식이 생쥐에서 혈중 지질농도 개선에 미치는 영향)

  • Hwang-Bo, Jeon;Jang, Kyung Ok;Chung, Hayoung;Park, Jong-Hwa;Lee, Tae Hoon;Kim, Jiyoung;Chung, In Sik
    • The Korean Journal of Food And Nutrition
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    • v.29 no.6
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    • pp.998-1007
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    • 2016
  • The objective of this study was to investigate the effects of a lettuce (Lactuca sativa L.) extract on the inflammation of human umbilical vein endothelial cell (HUVEC) and blood lipid improvement in hypercholesterolemic mice fed a high cholesterol diet. The lettuce extract (100% ethanol extract) inhibited the expression of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 in HUVEC treated with tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$). The lettuce extract suppressed the adhesion of THP-1 to TNF-${\alpha}$-treated HUVEC. The lettuce extract decreased the TNF-${\alpha}$-stimulated production of proinflammatory cytokine interleukin-6, interleukin-8 and chemokine monocyte chemotactic protein 1. In hypercholesterolemic mice, the lettuce extract reduced serum total cholesterol, triglyceride, and low-density lipoprotein-cholesterol level, while the lettuce extract elevated high-density lipoprotein-cholesterol level, resulting in the decrease of atherogenic index and cardiac risk factor level. These results suggested that lettuce extract can be an useful resource to show an anti-inflammatory effect and improve lipid metabolism.

Expression and Localization of 20α-Hydroxysteroid Dehydrogenase in Immature Pig Testis (미니돼지 정소에서 20α-HSD의 발현 및 특성화)

  • Kim, Jeong-Soo;Seong, Hun-Ki;Byambaragchaa, Munkhzaya;Sim, Bo-Woong;Her, Chang-Gi;Kang, Myung-Hwa;Min, Kwan-Sik
    • Journal of Life Science
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    • v.27 no.7
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    • pp.739-745
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    • 2017
  • In all mammalian species, progesterone is essential in the preparation for and maintenance of pregnancy. $20{\alpha}$-hydroxysteroid dehydrogenase ($20{\alpha}$-HSD) predominantly converts progesterone into its biologically inactive form $20{\alpha}$-hydroxyprogesterone ($20{\alpha}$-OHP), and plays a crucial role in the termination of pregnancy and initiation of parturition. In this study, we characterized the expression and localization of $20{\alpha}$-HSDinthe testis of MediKinetics $Micropigs^{(R)}$. The testes were collected at days 6, 9, 12, 18, and 21 after birth. The $20{\alpha}$-HSD mRNA was found to be expressed in the testis at day 6 after birth by RT-PCR. The highest level of mRNA expression in the testis was detected on day 21 after birth. However, the mRNA was not detected in the placenta after parturition. Western blot for $20{\alpha}$-HSD reveal that the specific 37-kDa band was detected in immature pig testis. However, this band was not detected in testis tissue at day 6 after birth. In the immunohistochemical analysis of the testis, $20{\alpha}$-HSD was detected in the Sertoli cells and Leydig cells. Taken together, our study shows for the first time that the $20{\alpha}$-HSD mRNA and protein are expressed in pig testis after birth. Further investigation is required to elucidate the functional mechanisms of $20{\alpha}$-HSD in pig testis after birth.

The Decreased Expression of Fbxw7 E3 Ligase Mediated by Cancer Upregulated Gene 2 Confers Cancer Stem Cell-like Phenotypes (CUG2 유전자에 의하여 감소된 FBXW7 E3 ligase 발현이 유사-종양줄기세포 표현형을 유도)

  • Yawut, Natpaphan;Kim, Namuk;Budluang, Phatcharaporn;Cho, Il-Rae;Kaowinn, Sirichat;Koh, Sang Seok;Kang, Ho Young;Chung, Young-Hwa
    • Journal of Life Science
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    • v.32 no.4
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    • pp.271-278
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    • 2022
  • The detailed mechanism by which cancer upregulated gene 2 (CUG2) overexpression induces cancer stem cell-like phenotypes is not fully understood. The downregulation of FBXW7 E3 ligase, a tumor suppressor known for its proteolytic regulation of oncogenic proteins such as cyclin E, c-Myc, Notch, and Yap1, has been frequently reported in several types of tumor tissues, including those in the large intestine, cervix, and stomach. Therefore, we investigated whether FBXW7 is involved in CUG2-induced oncogenesis. In this study, the decreased expression of FBXW7 was examined in human lung adenocarcinoma A549 (A549-CUG2) and human bronchial BEAS-2B cells (BEAS-CUG2) overexpressing CUG2 and compared with control cells stably expressing an empty vector (A549-Vec or BEAS-Vec). Treatment with MG132 (a proteosome inhibitor) prevented the degradation of FBXW7 and Yap1 proteins, which are substrates of the FBXW7 E3 ligase. To address the role of Fbxw7 in the development of cancer stem cell (CSC) phenotypes, we suppressed Fbxw7 protein levels using its siRNA. We observed that decreased levels of FBXW7 enhanced cell migration, invasion, and spheroid size and number in A549-Vec and BEAS-Vec cells. The enforced expression of FBXW7 produced the opposite results in A549-CUG2 and BEAS-CUG2 cells. Furthermore, the downregulation of FBXW7 elevated the activities of EGFR, Akt, and ERK1/2 and upregulated β-catenin, Yap1, and NEK2, while the enforced expression of FBXW7 generated the opposite results. We thus propose that FBXW7 downregulation induced by CUG2 confers CSC-like phenotypes through the upregulation of both the EGFR-ERK1/2 and β-catenin-Yap1-NEK2 signaling pathways.

Protective Effects of Korean Panax Ginseng Extracts against TCDD-induced Toxicities in Rat (랫드에서 TCDD 투여에 의해 유도된 생체독성의 고려홍삼 추출물에 의한 억제 효과)

  • Choi, Soo-Jin;Sohn, Hyung-Ok;Shin, Han-Jae;Hyun, Hak-Cheol;Lee, Dong-Wook;Song, Yong-Bum;Lee, Soo-Hyun;Gang, Dong-Ho;Lim, Hak-Seob;Lee, Cheol-Won;Moon, Ja-Young
    • Journal of Ginseng Research
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    • v.32 no.4
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    • pp.382-389
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    • 2008
  • To achieve a better understanding of protective effects of water extracts of Panax ginseng against TCDD-induced toxicities, we monitored physiological and clinical changes in rat for 4 weeks after administrations of each Panax Ginseng extract or TCDD, and co-administration of the two materials. For this study, 120 male Sprague-Dawley (SD) rats weighing 190-210 g each (8 weeks old) were divided into four groups: TCDD-administered, co-administered group with TCDD and ginseng extract, ginseng extract-administered, and control group. The TCDD-administered group received single dose of TCDD in a corn oil vehicle ($25\;{\mu}g/kg$ body weight) by intraperitoneal administration on Day 1. The Panax ginseng extracts-administered group received intraperitoneally 100 mg/kg body weight every other day for one month. For the co-administered group with TCDD and ginseng extracts, Panax ginseng extracts were intraperitoneally administered to rats at 100 mg/kg body weight every other day for one month after a single intraperitoneal dose of $25\;{\mu}g$ of TCDD/kg body weight on Day 1. Panax ginseng extracts attenuated the mortality induced by TCDD administration. The extracts also slightly attenuated the TCDD-induced body weight loss. Administration of TCDD alone increased liver weight at 2, 5, and 16 days after administration of TCDD. Administration of Panax ginseng extracts rather decreased liver weight through whole the experimental period, but which was statistically insignificant. Administration of TCDD alone at $25\;{\mu}g/kg$ body weight increased both serum enzyme activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) at 32 days, indicating that liver damage occurred maximally at that time. Ginseng extract administration caused insignificant changes in serum ALT, but gradually decreased in AST as the exposure time increased. Coadministration of TCDD and ginseng extracts caused serum AST activity to significant recovery to normal value at 16 days and 32 days after exposure to TCDD. The extracts also significantly decreased the TCDD-induced ALT activity after 16 days of TCDD administration. These results suggest that Panax ginseng extracts may possess a protective effect against TCDD-induced toxicities including hepatotoxicity in rats.

Substituting Bakery Waste for Barley Grains in Fattening Diets for Awassi Lambs

  • Hindiyeh, M.Y.;Haddad, S.G.;Haddad, S.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.11
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    • pp.1547-1551
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    • 2011
  • Bakery waste (BW) is much cheaper than barley (20 to 40% the price of barley). Bakery waste and barley grain have similar chemical composition; they contain 99 and 97% organic matter (OM), 1.1 and 1.8% fat, 18 and 15% neutral detergent fibre (NDF) and 14.0 and 14.5% crude protein (CP), respectively (DM basis). The objectives of this study were to determine the effect of substituting BW for barley grain in high concentrate fattening diets for lambs on nutrient intake, growth and carcass characteristics. Forty Awassi lambs (21.75${\pm}$1 kg) weaned at the age of 65 days were assigned randomly to four experimental fattening diets differing in BW ratio in a completely randomized design. The control diet (CON) contained 20, 60, 11, 7, and 2% (DM basis) wheat straw, barley grain, soybean meal, corn grain, and minerals and vitamin mix, respectively. Bakery waste substituted barley grain by 10, 20 and 30% of the diet DM in the LBW, MBW and HBW diets, respectively. The experiment lasted for 56 days. Dry matter intake (DMI) decreased (p<0.05) in LBW diet compared to the CON diet by approximately 10%. No further reduction in DMI was observed with the higher substitution levels. Metabolizable energy intake for the CON diet (3.6 Mcal/d) was also reduced (p<0.05) compared with LBW, MBW and HBW diets (3.4, 3.4 and 3.3 Mcal/d, respectively). Final body weight for lambs fed the CON diet (34.8 kg) was higher (p<0.05) compared with lambs fed the LBW, MBW and HBW diets (30.6, 32.0 and 31.1 kg, respectively). Growth rate for lambs fed the CON diet (232 g/d) was also higher (p<0.05) compared to lambs fed the LBW, MBW and HBW diets (170, 189, and 167 g/d, respectively). Feed to gain ratio was higher (p<0.05) for lambs fed the LBW, MBW and HBW diets (7.2, 6.6 and 7.3, respectively) compared with lambs that consumed the CON diet (5.7). Body weight gain cost was reduced by approximately 8% by the MBW and HBW diets as compared with the CON diet. Dressing percentage, full gut weight, empty gut weight and liver weights were all unaffected by the BW addition to the diets and averaged 48.9%, 6.8 kg, 2.8 kg and 0.444 kg, respectively. However, fat tail weight was increased (p<0.05) with the higher levels of the BW inclusion. In conclusion, substituting BW for barley grain reduced DMI and growth performance. However, when BW substituted barley grain at the 20 and 30% of the diet DM, body weight gain cost was reduced by approximately 8%.

Wolbachia-mediated Reproductive Alterations in Arthropod Hosts and its use for Biocontrol Program (볼바키아 세균에 의한 절지동물 기주의 생식적 변화와 생물적방제 프로그램에 이용 방안)

  • Rostami, Elahe;Madadi, Hossein;Abbasipour, Habib;Sivaramakrishnan, Shiva
    • Korean journal of applied entomology
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    • v.55 no.2
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    • pp.177-188
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    • 2016
  • The alpha-proteobacterium Wolbachia is one of the most important intracellular symbionts of arthropods. This Gram-negative bacterium is involved in many biological processes and is currently considered as a potential tool for biological control. Wolbachia is a cytoplasmic bacterium, maternally transferred through generations, and to facilitate its success, it has evolved several strategies that manipulate its host reproductive system to increase the number of infected individuals in the host population. The variety of Wolbachia was first recognized using genes wsp, 16S rRNA, ftsZ, gltA and groEL as molecular markers while strain genotypes of Wolbachia are determined of Multilocus sequence typing (MLST) and sequence of amino acid in region, hyper variable regions (HVRs) in protein WSP. Possible uses of the bacteria and their predominant phenotypes in control programs for agricultural pests and human disease vectors have been considered. Phenotypes are known to induce cytoplasmic incompatibility (CI), parthenogenesis induction (PI), feminization (F) and male killing (MK). Finally, applications of the bacterium in control programs of agricultural and medical insect pests have been discussed.

Comparison of Metabolic Pathways of Less Orthologous Prokaryotes than Mycoplasma genitalium (Mycoplasma genitalium 보다 보존적 유전자 수가 작은 원핵생물들의 대사경로 비교)

  • Lee, Dong-Geun
    • Journal of Life Science
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    • v.28 no.3
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    • pp.369-375
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    • 2018
  • Mycoplasma genitalium has 367 conserved genes and the smallest genome among mono-culturable prokaryotes. Conservative metabolic pathways were examined among M. genitalium and 14 prokaryotes, one hyperthermophilic exosymbiotic archaeon Nanoarchaeum equitans and 13 intracellular eubacteria of plants or insects, with fewer conserved genes than M. genitalium. They have 11 to 71 metabolic pathways, however complete metabolic pathways ranged from 1 to 24. Totally, metabolic pathway hole is very high due to the lack of 45.8% of the enzymes required for the whole metabolic pathways and it could be suggested that the shared metabolic pathway with the host's enzyme would work or the essential substances are host dependent. The number of genes necessary for mass transfer through the cell membrane is also very low, and it may be considered that the simple diffusion or the protein of the host will function in the cell membrane of these prokaryotes. Although the tRNA charging pathway was distributed in all 15 prokaryotes, each has 5-20 tRNA charging genes. This study would give clues to the understanding of the metabolic pathways of intracellular parasitic bacteria of plant and endosymbiotic bacteria of insects, and could provide basic data for prevention of crop damage, development of insect pests and human medicines.

Anti-inflammation Effect of Gynura Procumbens extract (명월초 추출물의 항염 효과에 대한 연구)

  • Jeon, Hyeong-Ju;Kwon, Hye-Jin
    • Journal of Digital Convergence
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    • v.14 no.10
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    • pp.515-520
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    • 2016
  • This study conducted the research about the positive effects of Gynura Procumbens on preventing dermatitis and infectious diseases the contemporary people have. To do it, this study measured the concentration of NF-kB, the extract of Gynura Procumbens. In this regard, cytosol, the extract of Gynura Procumbens, and nucleus went through the separation process. The concentration of major NF-kB among various factors to control inflammation was measured with the use of HDF cell. Regarding RAW264.7 cell, the amounts of NO to play an important role in reacting the skin immune system as the media of neural transmission were analyzed. The outcome about the extracts of Gynura Procumbens injected show that it can be expected that as the NF-kB protein and mRNA band were reduced, Gynura Procumbens would have anti-inflammatory effects that could contribute to preventing dermatitis and diseases. In addition, the extracts of Gynura Procumbens have significantly reduced NO with their concentration increasing. In other words, Gynura Procumbens are considered to regulate dependently the production of NO in the concentration of extracts. Thus there is an expectation that the more intensive research would be conducted to heal dermatitis. And it is deemed that Gynura Procumbens would be used as materials for cosmetics as well as foods that the contemporary people can widely consume if a more careful research about anti-inflammatory effects would be sustained on a human bodies' clinical level.