• Title/Summary/Keyword: Synthetic biology

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The Relationship between Virginiae Butanolide C(VB-C) and Receptor in Virginiamycin Production (Virginiamycin 생산유도에 관여하는 Virginiae Butanolide C(VB-C) 및 Receptor의 상관관계)

  • Kim, Hyun-Soo;Hyun, Ji-Sook;Yu, Tae-Shick
    • Microbiology and Biotechnology Letters
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    • v.24 no.1
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    • pp.59-66
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    • 1996
  • Virginiae butanolide C(VB-C) is one of the butyrolactone autoregulators, which triggers the productin of virginiamycin in Streptomyces virginiae. To further understand the mechanism of virginiamycin induction, we isolated three mutants from S. virginiae by N-methyl-N'-nitrosoguanidine (NTG) treatment. The characteristics of the three mutants were confirmed as follows: the mutant No. 1 delayed the production of the VB-C, receptor and antibiotics; the mutant No.3 hyperproduced receptor; the mutant No.4 failed to produce the VB-C. The addition of synthetic VB-C couldn't induce the production of antibiotics in the mutant No.1 due to delayed production of receptor, could provoke the production of larger amount of antibiotics than parental wild type strain in the mutant No.3 due to the presence of large amount of receptor, and could induce production of very small amount of antibiotics in the mutant No.4 due to the absence of VB-C. Antimicrobial spectrum and HPLC analysis of the mutant No.1 and No.3 suggested that the VB-C might have a specific ability to induce the production of virginiamycin M and S. These results imply that the VB-C has an ability to trigger the production of virginiamycin under receptor existence in S. virginiae.

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A Novel All-trans Retinoid Acid Derivative Induces Apoptosis in MDA-MB-231 Breast Cancer Cells

  • Wang, Bei;Yan, Yun-Wen;Zhou, Qing;Gui, Shu-Yu;Chen, Fei-Hu;Wang, Yuan
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.24
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    • pp.10819-10824
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    • 2015
  • Aims: To explore the effect and probable mechanism of a synthetic retinoid 4-amino-2-tri-fluoromethylphenyl ester (ATPR) on apoptosis of MDA-MB-231 breast cancer cells. Materials and Methods: MTT assays were performed to measure the proliferation of MDA-MB-231 cells treated with different concentrations of all-trans retinoic acid (ATRA) and ATPR. Morphologic changes were observed by microscopy. The apoptosis rates and cell cycling of MDA-MB-231 cells treated with ATRA or ATPR were assessed using flow cytometry analysis. Expression of retinoic acid receptor and phosphorylation of ERK, JNK, p38 proteins were detected by Western blotting. Results: Treatment of the cells with the addition of $15{\mu}mol/L$ ATPR for 48 h clearly demonstrated reduced cell numbers and deformed cells, whereas no changes in the number and morphology were observed after treatment with ATRA. The apoptosis rate was 33.2% after breast cancer MDA-MB-231 cells were treated by ATPR ($15{\mu}mol/L$) whereas ATRA ($15{\mu}mol/L$) had no apoptotic effect. ATPR inhibited the phosphorylation of ERK, JNK, and p38 while ATRA had no significant effect. ATPR inhibited the expression of BiP and increased the expression of Chop at the protein level compared with control groups, ATRA and ATPR both decreased the protein expression of $RXR{\alpha}$, ATPR reduced the protein expression of $RAR{\beta}$ and $RXR{\beta}$ while ATRA did not decrease $RAR{\beta}$ or $RXR{\beta}$. Conclusions: ATPR could induce apoptosis of breast cancer MDA-MB-231 cells, possible mechanisms being binding to $RAR{\beta}/RXR{\beta}$ heterodimers, then activation of ER stress involving the MAPK pathway.

Protoplast Formation and Regeneration from Mycelia of Phytophthora capsici (Phytophthora capsici의 균사체(菌絲體)로부터 원형질체(原形質體) 형성(形成)과 재생(再生))

  • Yi, Seung-Youn;Kim, Young-Jin;Hwang, Byung-Kook
    • The Korean Journal of Mycology
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    • v.21 no.1
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    • pp.1-8
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    • 1993
  • ABSTRACT: Factors responsible for protoplast formation and regeneration of Phytophthora capsici were examined. Protoplasts were successfully liberated from the mycelial culture by digestion for 6-9 hrs with Novozym 234 in 0.35 M $CaCl_2$, (pH 5.7) as osmotic stabilizer. Young rapidly-growing mycelium (24 hrs old) showed highest protoplast yields. High concentrations of Novozym 234 were effective in releasing protoplasts from the mycelium. The combination of 0.4 M mannitol and 0.1 M $CaCl_2$ was optimal osmotic stabilizers for protoplast regeneration. The synthetic Henninger media containing all nutritional elements gave the best regeneration rate. The protoplast regeneration was greatly inhibited in the media which were not supplement with amino acids or ${\beta}-sitosterol$. Certain amino acids such as L-aspartic acid and L-glutamic acid remarkably enhanced protoplast regeneration. However, the addition of microelements did not affect protoplast regeneration.

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Investigation on Cultural Characteristics of Mycelial Growth by Cordyceps scarabaeicola (풍뎅이동충하초(Cordyceps scarabaeicola)의 균사 생장 조건 규명)

  • Lee, Jae-Keun;Choi, Young-Sang;Sung, Jae-Mo
    • The Korean Journal of Mycology
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    • v.28 no.2
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    • pp.81-87
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    • 2000
  • The optimal synthetic medium, temperature and pH range for the mycelial growth of Cordyceps scarabaeicola were PDA, $25^{\circ}C$, and pH $8.0{\sim}8.5$, respectively. The effects of carbon sources, nitrogen sources and mineral salts on mycelial growth were superior to lactose, ammonium phosphate and $CaCl_2$, respectively. Liquid spawn culture was selected out BM(brown sugar 3%, soybean flour 0.3%, potassium phosphate 0.05%, magnesium sulfate 0.05%) medium.

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Efficient Elicitation of Ginsenoside Biosynthesis in Cell Cultures of Panax notoginseng by Using Self-chemically-synthesized Jasmonates

  • Wang Wei;Zhao Zhen-Jiang;Xu Yufang;Qian Xu hong;Zhong Jian-Jiang
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.2
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    • pp.162-165
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    • 2005
  • A series of fluorine and hydroxyl containing jasmonate derivatives, which were chemically synthesized in our institute, were investigated for their effects on the biosynthesis and heterogeneity of ginsenosides in suspension cultures of Panax notoginseng cells. Com-pared to the control (without addition of elicitors), $100{\mu}M$ of each of the jasmonate was added on day 4 to the suspension cultures of P. notoginseng cells. It was observed that, jasmonates greatly enhanced the ginsenoside content and the ratio of Rb group to Rg group (i.e. $(Rb_1\;+\;Rd)/(Rg_1\;+\;Re)$ in the P. notoginseng cells. Some of the synthetic jasmonates, such as pentafluoropropyl jasmonate (PFPJA), 2-hydroxyethyl jasmonate (HEJA) and 2-hydroxye-thoxyethyl jasmonate (HEEJA), could promote the ginsenoside content to $2.55\;\pm\;0.11,\;3.65\;\pm\;0.13\;and\;2.94\;\pm\;0.06$mg/100 mg DW, respectively, compared to that of $0.64\;\pm\;0.06$mg/100 mg DW for the control and $2.17\;\pm\;0.04$ mg/100 mg DW by the commercially available methyl jasmonate (MJA); and they could change the respective Rb:Rg ratio to $1.60\;\pm\;0.04,\;1.87\;\pm\;0.01\;and\;1.56\;\pm\;0.05$, compared to that of $0.47\;\pm\;0.01$ for the control and $1.42\;\pm\;0.06$ by MJA. The results suggest that suitable esterification of MJA with fluorine or hydroxyl group could in-crease the elicitation activity to induce plant secondary metabolism. The information obtained from this study is useful for hyper-production of heterogeneous products by plant cell cultures.

KMS99220 Exerts Anti-Inflammatory Effects, Activates the Nrf2 Signaling and Interferes with IKK, JNK and p38 MAPK via HO-1

  • Lee, Ji Ae;Kim, Dong Jin;Hwang, Onyou
    • Molecules and Cells
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    • v.42 no.10
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    • pp.702-710
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    • 2019
  • Neuroinflammation is an important contributor to the pathogenesis of neurodegenerative disorders including Parkinson's disease (PD). We previously reported that our novel synthetic compound KMS99220 has a good pharmacokinetic profile, enters the brain, exerts neuroprotective effect, and inhibits $NF{\kappa}B$ activation. To further assess the utility of KMS99220 as a potential therapeutic agent for PD, we tested whether KMS99220 exerts an anti-inflammatory effect in vivo and examined the molecular mechanism mediating this phenomenon. In 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-treated mice, oral administration of KMS99220 attenuated microglial activation and decreased the levels of inducible nitric oxide synthase and interleukin 1 beta ($IL-1{\beta}$) in the nigrostriatal system. In lipopolysaccharide (LPS)-challenged BV-2 microglial cells, KMS99220 suppressed the production and expression of $IL-1{\beta}$. In the activated microglia, KMS99220 reduced the phosphorylation of $I{\kappa}B$ kinase, c-Jun N-terminal kinase, and p38 MAP kinase; this effect was mediated by heme oxygenase-1 (HO-1), as both gene silencing and pharmacological inhibition of HO-1 abolished the effect of KMS99220. KMS99220 induced nuclear translocation of the transcription factor Nrf2 and expression of the Nrf2 target genes including HO-1. Together with our earlier findings, our current results show that KMS99220 may be a potential therapeutic agent for neuroinflammation-related neurodegenerative diseases such as PD.

Immersion in sea cucumber's steroid extract to increase male production of juvenile freshwater crayfish

  • Gregorius Nugroho Susanto;Endang Linirin Widiastuti;Tri Rustanti;Sutopo Hadi
    • Fisheries and Aquatic Sciences
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    • v.26 no.1
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    • pp.48-57
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    • 2023
  • One of the ways to increase the production for aquaculture is through the cultivation of monosexuals by ensuring genital reversal from which energy for reproduction is diverted towards growth. Masculinization has been identified as one of the most prominent techniques, where sex development was directed from female to male. This approach only altered the phenotype and not the genotype. The red claw crayfish (Cherax quadricarinatus) was a relatively new commercial commodity, and the males were known to grow faster than females. Hence, it was proposed to use monocultures comprising an all-male population to increase yield using steroid hormone, synthetic 17α-methyltestosterone. However, this technique generated residues that detrimentally affect human health, the environment, and cultivated organisms. Therefore, finding new safe natural steroid sources was essential, and one of which is exploring of natural hormones extracted from the viscera of sea cucumbers (Holothuria scabra Jaeger). This study focused on the determination of male formation and testosterone levels among juvenile crayfish, after immersing in sea cucumber steroid extract (SCSE). A completely random design with factorial was used with two variables, encompassing the varied doses (0, 2, 4 mg/L, 2 mg/L 17α-methyl testosterone as control group) and immersion times of 18 and 30 h. The result showed the dose-dependent ability of SCSE increase the male genital formation and promote the testosterone level of juvenile crayfish. In addition, the testosterone was influenced by dose and immersion duration time, with the highest level of testosterone observed in treatments of 4 mg/L SCSE with 30 h immersion was 0.248 ng/mL, while the male percentage was 77%. In conclusion, the combination of dose and immersion time significantly affected growth and testosterone levels.

Immunomodulatory Response Induced by Ginseng

  • Kumar, Ashok
    • Journal of Ginseng Research
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    • v.27 no.3
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    • pp.115-119
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    • 2003
  • There has been continuing interest in the development of synthetic and natural compounds that modify the immune response particularly for the treatment of AIDS and cancer. During the past fifty years, numerous scientific studies have been published on ginseng. Modem human studies have investigated preventive effect of ginseng on several kinds of cancer, its long term immunological effect on HIV patients, its effect on cell mediated immune functions in healthy volunteers. Similarly non clinical studies on animal model system have studied the chemopreventive action of ginseng on cancer and immunological properties of ginseng. The precise mechanism of action of ginseng, however, not clearly understood. Considering its wide-ranging therapeutic effects, this study is being undertaken to elucidate the general mode of action of ginseng, especially to test our hypothesis that its biological action may be mediated by the immune system.

Sequence Verification of Synthetic Oligonucleotides by Exonuclease Digestion and Matrix Assisted Laser Desorption Ionization Mass Spectrometry

  • Kim, Jin-Sung;Jang, Jung-Suk;Choi, Jong-Soon;Chang, Yoon-Seok
    • BMB Reports
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    • v.29 no.2
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    • pp.122-126
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    • 1996
  • A series of oligonucleotides were synthesized by automatic DNA synthesizer. The purity of crude products was checked and their molecular weights determined by matrix assisted laser desorption ionization mass spectrometry (MALDI-MS) with an accuracy of better than 0.05% deviation even without using an internal standard. This mass determining technology in combination with partial digestion of oligonucleotides by 5'- and 3'-exonuclease provides a straightforward and simple method to obtain sequence information of oligonucleotides. The extension of this technology to the sequencing of modified oligonucleotides and genomic DNA and RNA might become possible.

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Prenatal Development of Tvrosine Hlrdroxvlase-Containing Neurons in the Rat Brain (흰쥐 태아 뇌에서 TH 신경세포의 초기발생과정)

  • 이영기;장가용
    • The Korean Journal of Zoology
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    • v.36 no.2
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    • pp.209-222
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    • 1993
  • The present study attempts to explore the first appearance 8nd subsequent development of %cosine hvdroxvlase (TH)-containing neurons in the rat brain from embryonic day (E) 10.5 to the neonate. To increase the senti노ivity and resolution power, a double bridge peroxidase-anti-peroxidase technique was employed for cellular localization of TH. In situ hybridization histochemistw with synthetic TH oligomer (30-mer) codinB TH was also used to detect TH mRNA. TH-containing neurons were first detected at E11.5 in the intermediate zone of prosencepha1on and mesencephalon. At this stage, TH-immunoreactive neurons were small, ovoid bee and emitted their fibres into their immediate surroundings. From this stage, TH-immunoreactive neurons increased in their number and underwent migration and cell differentiation. At E15.5, the distribution pattern of the maior groups of TH neurons was similar to that of adult catecholaminergic groups, and at E19.5 the external laver of median eminence showed TH-immunoreactive processes.

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