• Title/Summary/Keyword: overproduction.

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Maximization of Poly-$\beta$-Hydroxybutyrate Accumulation by Potassium Limitation in Methylobacterium organophilum and Its Related Metabolic Analysis

  • Kim, Seon-Won;Kim, Pil;Kim, Jung-Hoe
    • Journal of Microbiology and Biotechnology
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    • v.9 no.2
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    • pp.140-146
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    • 1999
  • When methanol was the sole carbon source, Methylobacterium organophilum NCIB 11278, a facultative methylotroph, accumulated Poly-$\beta$-hydroxybutyrate (PHB) as 59% (w/w) of dry cell weight under potassium limitation, 37% under sulfate limitation, and 33% under nitrogen limitation. Based on a stoichiometric analysis of PHB synthesis from methanol, it was suspected that PHB synthesis is accompanied by the overproduction of energy, either 6-10 ATP and 1 $FADH_2$ or 6 ATP and 3 NADPH to balance the NADH requirement, per PHB monomer. This was confirmed by observation of increased intracellular ATP levels during PHB accumulation. The intracellular ATP with limited potassium, sulfate, and ammonium increased to 0.185, 0.452, and 0.390 $\mu$moles ATP/g Xr (residual cell mass) during PHB accumulation, respectively. The intracellular ATP level under potassium limitation was similar to that when there was no nutrient limitation and no PHB accumulation, 0.152- 0.186 $\mu$moles ATP/g Xr. We propose that the maximum PHB accumulation observed when potassium was limited is a result of the energy balance during PHB accumulation. Microorganisms have high energy requirements under potassium limitation. Enhanced PHB accumulation, in ammonium and sulfate limited conditions with the addition of 2,4-dinitrophenol, which dissipates surplus energy, proves this assumption. With the addition of 1 mM of 2,4-dinitrophenol, the PHB content increased from 32.4% to 58.5% of dry cell weight when nitrogen limited and from 15.1 % to 31.0% of dry cell weight when sulfate limited.

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Melanin Bleaching and Melanogenesis Inhibition Effects of Pediococcus acidilactici PMC48 Isolated from Korean Perilla Leaf Kimchi

  • Kim, Sukyung;Seo, Hoonhee;Mahmud, Hafij Al;Islam, Md Imtiazul;Sultana, Omme Fatema;Lee, Youngkyoung;Kim, Minhee;Song, Ho-Yeon
    • Journal of Microbiology and Biotechnology
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    • v.30 no.7
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    • pp.1051-1059
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    • 2020
  • Overproduction and accumulation of melanin in the skin will darken the skin and cause skin disorders. So far, components that can inhibit tyrosinase, a melanin synthase of melanocytes, have been developed and used as ingredients of cosmetics or pharmaceutical products. However, most of existing substances can only inhibit the biosynthesis of melanin while melanin that is already synthesized and deposited is not directly decomposed. Thus, their effects in decreasing melanin concentration in the skin are weak. To overcome the limitation of existing therapeutic agents, we started to develop a substance that could directly biodegrade melanin. We screened traditional fermented food microorganisms for their abilities to direct biodegrade melanin. As a result, we found that a kimchi-derived Pediococcus acidilactici PMC48 had a direct melanin-degrading effect. This PMC48 strain is a new strain, different from P. acidilactici strains reported so far. It not only directly degrades melanin, but also has tyrosinase-inhibiting effect. It has a direct melanin-decomposition effect. It exceeds existing melanin synthesis-inhibiting technology. It is expected to be of high value as a raw material for melanin degradation drugs and cosmetics.

$Mycoplasma$ $pneumoniae$ pneumonia in children

  • Youn, You-Sook;Lee, Kyung-Yil
    • Clinical and Experimental Pediatrics
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    • v.55 no.2
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    • pp.42-47
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    • 2012
  • $Mycoplasma$ $pneumoniae$ (MP), the smallest self-replicating biological system, is a common cause of upper and lower respiratory tract infections, leading to a wide range of pulmonary and extra-pulmonary manifestations. MP pneumonia has been reported in 10 to 40% of cases of community-acquired pneumonia and shows an even higher proportion during epidemics. MP infection is endemic in larger communities of the world with cyclic epidemics every 3 to 7 years. In Korea, 3 to 4-year cycles have been observed from the mid-1980s to present. Although a variety of serologic assays and polymerase chain reaction (PCR) techniques are available for the diagnosis of MP infections, early diagnosis of MP pneumonia is limited by the lack of immunoglobulin (Ig) M antibodies and variable PCR results in the early stages of the infection. Thus, short-term paired IgM serologic tests may be mandatory for an early and definitive diagnosis. MP infection is usually a mild and self-limiting disease without specific treatment, and if needed, macrolides are generally used as a first-choice drug for children. Recently, macrolide-resistant MP strains have been reported worldwide. However, there are few reports of apparent treatment failure, such as progression of pneumonia to acute respiratory distress syndrome despite macrolide treatment. The immunopathogenesis of MP pneumonia is believed to be a hyperimmune reaction of the host to the insults from MP infection, including cytokine overproduction and immune cell activation (T cells). In this context, immunomodulatory treatment (corticosteroids or/and intravenous Ig), in addition to antibiotic treatment, might be considered for patients with severe infection.

3,4,5-Trihydroxycinnamic Acid Inhibits LPS-Induced iNOS Expression by Suppressing NF-${\kappa}B$ Activation in BV2 Microglial Cells

  • Lee, Jae-Won;Bae, Chang-Jun;Choi, Yong-Jun;Kim, Song-In;Kim, Nam-Ho;Lee, Hee-Jae;Kim, Sung-Soo;Kwon, Yong-Soo;Chun, Wan-Joo
    • The Korean Journal of Physiology and Pharmacology
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    • v.16 no.2
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    • pp.107-112
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    • 2012
  • Although various derivatives of caffeic acid have been reported to possess a wide variety of biological activities such as neuronal protection against excitotoxicity and anti-inflammatory property, the biological activity of 3,4,5-trihydroxycinnamic acid (THC), a derivative of hydroxycinnamic acids, has not been clearly examined. The objective of the present study is to evaluate the anti-inflammatory effects of THC on lipopolysaccharide (LPS)-stimulated BV2 microglial cells. THC significantly suppressed LPS-induced excessive production of nitric oxide (NO) and expression of iNOS, which is responsible for the production of iNOS. THC also suppressed LPS-induced overproduction of pro-inflammatory cytokines such as IL-$1{\beta}$and TNF-${\alpha}$ in BV2 microgilal cells. Furthermore, THC significantly suppressed LPS-induced degradation of $I{\kappa}B$, which retains NF-${\kappa}B$ in the cytoplasm. Therefore, THC attenuated nuclear translocation of NF-${\kappa}B$, a major pro-inflammatory transcription factor. Taken together, the present study for the first time demonstrates that THC exhibits antiinflammatory activity through the suppression of NF-${\kappa}B$ transcriptional activation in LPS-stimulated BV2 microglial cells.

The anti-inflammatory effect of Indonesian Areca catechu leaf extract in vitro and in vivo

  • Lee, Kang Pa;Sudjarwo, Giftania Wardani;Kim, Ji-Su;Dirgantara, Septrianto;Maeng, Won Jai;Hong, Heeok
    • Nutrition Research and Practice
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    • v.8 no.3
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    • pp.267-271
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    • 2014
  • BACKGROUND/OBJECTIVES: Overproduction of nitric oxide (NO) by the inducible nitric oxide synthase (iNOS) enzyme can cause inflammation. Cyclooxygenase-2 (COX-2) is also involved in the inflammatory response through regulation of nuclear factor-kappa B $NF-{\kappa}B$(). Areca catechu is one of the known fruit plants of the Palmaceae family. It has been used for a long time as a source of herbal medicine in Indonesia. In this study, we explored the effect of Indonesian Areca catechu leaf ethanol extract (ACE) in lipopolysaccharide (LPS)-induced inflammation and carrageenan-induced paw edema models. Recently, this natural extract has been in the spotlight because of its efficacy and limited or no toxic side effects. However, the mechanism underlying its anti-inflammatory effect remains to be elucidated. MATERIALS/METHODS: We measured NO production by using the Griess reagent, and determined the expression levels of inflammation-related proteins, such as iNOS, COX2, and $NF-{\kappa}B$, by western blot. To confirm the effect of ACE in vivo, we used the carrageenan-induced paw edema model. RESULTS: Compared to untreated cells, LPS-stimulated RAW 264.7 cells treated with ACE showed reduced NO generation and reduced iNOS and COX-2 expression. We found that the acute inflammatory response was significantly reduced by ACE in the carrageenan-induced paw edema model. CONCLUSION: Taken together, these results suggest that ACE can inhibit inflammation and modulate NO generation via downregulation of iNOS levels and $NF-{\kappa}B$ signaling in vitro and in vivo. ACE may have a potential medical benefit as an anti-inflammation agent.

The Effects of Ischemic Postconditioning on Myocardial Function and Nitric Oxide Metabolites Following Ischemia-Reperfusion in Hyperthyroid Rats

  • Zaman, Jalal;Jeddi, Sajjad;Ghasemi, Asghar
    • The Korean Journal of Physiology and Pharmacology
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    • v.18 no.6
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    • pp.481-487
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    • 2014
  • Ischemic postconditioning (IPost) could decrease ischemia-reperfusion (IR) injury. It has not yet reported whether IPost is useful when ischemic heart disease is accompanied with co-morbidities like hyperthyroidism. The aim of this study was to examine the effect of IPost on myocardial IR injury in hyperthyroid male rats. Hyperthyroidism was induced with administration of thyroxine in drinking water (12 mg/L) over a period of 21 days. After thoracotomy, the hearts of control and hyperthyroid rats were perfused in the Langendorff apparatus and subjected to 30 minutes global ischemia, followed by 120 minutes reperfusion; IPost, intermittent early reperfusion, was induced instantly following ischemia. In control rats, IPost significantly improved the left ventricular developed pressure (LVDP) and ${\pm}dp/dt$ during reperfusion (p<0.05); however it had no effect in hyperthyroid rats. In addition, hyperthyroidism significantly increased basal $NO_x$ (nitrate+nitrite) content in serum ($125.5{\pm}5.4{\mu}mol/L$ vs. $102.8{\pm}3.7{\mu}mol/L$; p<0.05) and heart ($34.9{\pm}4.1{\mu}mol/L$ vs. $19.9{\pm}1.94{\mu}mol/L$; p<0.05). In hyperthyroid groups, heart $NO_x$ concentration significantly increased after IR and IPost, whereas in the control groups, heart $NO_x$ were significantly higher after IR and lower after IPost (p<0.05). IPost reduced infarct size (p<0.05) only in control groups. In hyperthyroid group subjected to IPost, aminoguanidine, an inducible nitric oxide (NO) inhibitor, significantly reduced both the infarct size and heart $NO_x$ concentrations. In conclusion, unlike normal rats, IPost cycles following reperfusion does not provide cardioprotection against IR injury in hyperthyroid rats; an effect that may be due to NO overproduction because it is restored by iNOS inhibition.

Effect of KH-305 on the Nitric Oxide Synthase Activity and Erectile Dysfunction in Young Rats (KH-305 투여가 흰쥐 음경조직의 Nitric Oxide Synthase활성 및 Erectile Dysfunction에 미치는 영향)

  • Lee, Eun-Jeong;Kim, Hee-Seok;Kim, Byoung-Chul;Hwang, Sung-Wan;Hwang, Sung-Yeoun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.3
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    • pp.305-310
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    • 2007
  • This study was designed to investigate the effects of KH-305 on erectile dysfunction in young rats, via nitric oxide (NO)-cGMP pathways. After oral administration of the KH-305 mixture (50, 100, 200, 300 mg/kg) to young rats for 10 days, NOS and SOD protein expressions in penile tissue and testosterone in plasma were measured. cGMP degradation was also investigated using bovine vascular smooth muscle cells pretreated with an NO donor, S-nitroso-N-Acetylpenicillamine (SNAP). The penile expression levels of nNOS and eNOS-dependent NOS activities as well as SOD preventing oxidative stress by overproduction of NO were increased significantly. Also, the concentration of testosterone in the plasma was increased. In vitro, cGMP concen-trations were decreased dose dependently in the KH-305. These results suggest that KH-305 may be useful in erectile dysfunction.

Effect of Nutrients and Applications for the Overproduction of Polyoxins by Streptomyces Speies (Streptomyces 균주의 Polyoxins 생합성 증대를 위한 영양분 효과)

  • 김상호;주현유영제박영훈
    • KSBB Journal
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    • v.10 no.2
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    • pp.196-203
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    • 1995
  • The effects of majors nutrients and environmental factors on polyoxins biosysnthesis were examined for the overprodution of antifungal agent polyoxins using Streptomyces sp. 809-11. The cell mass at the exponential growth phase was increased when soluble starch was used, while the polyoxins biosynthesis was increased with the formation of filamentous mycelium when glucose was used as a carbon source. It was, therefore, recommendable to use both soluble starch and glucose simultaneously as carbon sources for the cell growth and polyoxins production. The high concentration of ammonium sulfate (151.4 mM) resulied in the improvement of polyoxins production. The optimal concentration of $K_2HPO_4$for polyoxins production was found to be 0.5mM. By applying fed-batch culture, polyoxins production was improved to about 2-folds compared to the result from the batch culture.

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Development and Characterization of Sporulation Mutants for Overexpression of Recombinant Protein of Bacillus subtilis (재조합 단백질 과발현을 위한 Bacillus snbtilis 포자형성 변이주의 개발 및 특성 분석)

  • 오민규;박승환김병기
    • KSBB Journal
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    • v.9 no.1
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    • pp.16-25
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    • 1994
  • Sporulation mutants of Bacillus subtilis were developed for overproduction of heterologous proteins. The strains spoOJ spoIIG, and spoOJ spoIIG double mutant were constructed from two pretense-delfted mutant (DB104). The vector containing aprE gene was integrated in the chromosome of each strain, then the morphology of each strain was observed by TEM (trasmission electron microscopy). The morphology of spoOJ mutant and spoIIG mutant coincides with the description of the previous reports, respectively. The sporulating cells of spoOJ SpoIIG double mutation resemble spoIIG mutant more similarly, but with a little rougher cell wall membrane. The spoOJ mutation in B. subtilis gives negative effect on aprE activity with only a decreased sporulation frequency. On the contrary spoIIG mutation increases the aprE activity twice with an undetectable sporulation frequency. In the case of spoOJ and spolIG, i. e. double mutation, the effect of spoOJ on aprE activity seems to be relieved and the double mutant shows more or less the same aprE activity compared to spoIIG mutant.

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Nitric Oxide as a Pro-apoptotic as well as Anti-apoptotic Modulator

  • Choi, Byung-Min;Pae, Hyun-Ock;Jang, Seon-Il;Kim, Young-Myeong;Chung, Hun-Taeg
    • BMB Reports
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    • v.35 no.1
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    • pp.116-126
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    • 2002
  • Nitric oxide (NO), synthesized from L-arginine by NO synthases, is a small, lipophilic, diffusible, highly reactive molecule with dichotomous regulatory roles in many biological events under physiological and pathological conditions. NO can promote apoptosis (pro-apoptosis) in some cells, whereas it inhibits apoptosis (anti-apoptosis) in other cells. This complexity is a consequence of the rate of NO production and the interaction with biological molecules such as metal ion, thiol, protein tyrosine, and reactive oxygen species. Long-lasting overproduction of NO acts as a pro-apoptotic modulator, activating caspase family proteases through the release of mitochondrial cytochrome c into cytosol, up-regulation of the p53 expression, and alterations in the expression of apoptosis-associated proteins, including the Bcl-2 family. However, low or physiological concentrations of NO prevent cells from apoptosis that is induced by the trophic factor withdrawal, Fas, $TNF{\alpha}$/ActD, and LPS. The anti-apoptotic mechanism is understood on the basis of gene transcription of protective proteins. These include: heat shock protein, hemeoxygenase, or cyclooxygenase-2 and direct inhibition of the apoptotic executive effectors caspase family protease by S-nitrosylation of the cysteine thiol group in their catalytic site in a cell specific way. Our current understanding of the mechanisms by which NO exerts both pro- and anti-apototic action is discussed in this review article.