• 제목/요약/키워드: bacterial hemoglobin

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Introduction of a Bacterial Hemoglobin Gene for Improving Bacterial Growth under Hypoxic Condition

  • Chung, Chung-Nam;Yoon, Suk-Ran;Jun, Woo-Jin;Shim, Sang-In;Park, In-Ho;Chung, Jin-Woong
    • 농업생명과학연구
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    • 제43권6호
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    • pp.77-84
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    • 2009
  • Using recombinant DNA technology, the vector system containing minimal fragment of a bacterial hemoglobin gene (vgb) was constructed. When this vector was inserted into Escherichia coli, the growth of the host was significantly improved in both viable cell counts and absorbance measurement, compared to that of the wild type strain. In addition, by minimizing the size of bacterial hemoglobin in the vector, the ability of vgb in growth improvement was augmented, due to the reduction of metabolic burden from the maintenance and replication of the plasmid. By using this system, it is expected that the growth of microorganisms can be improved even in the hypoxic condition.

Enhanced Production of ${\varepsilon}$-Caprolactone by Coexpression of Bacterial Hemoglobin Gene in Recombinant Escherichia coli Expressing Cyclohexanone Monooxygenase Gene

  • Lee, Won-Heong;Park, Eun-Hee;Kim, Myoung-Dong
    • Journal of Microbiology and Biotechnology
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    • 제24권12호
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    • pp.1685-1689
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    • 2014
  • Baeyer-Villiger (BV) oxidation of cyclohexanone to ${\varepsilon}$-caprolactone in a microbial system expressing cyclohexanone monooxygenase (CHMO) can be influenced by not only the efficient regeneration of NADPH but also a sufficient supply of oxygen. In this study, the bacterial hemoglobin gene from Vitreoscilla stercoraria (vhb) was introduced into the recombinant Escherichia coli expressing CHMO to investigate the effects of an oxygen-carrying protein on microbial BV oxidation of cyclohexanone. Coexpression of Vhb allowed the recombinant E. coli strain to produce a maximum ${\varepsilon}$-caprolactone concentration of 15.7 g/l in a fed-batch BV oxidation of cyclohexanone, which corresponded to a 43% improvement compared with the control strain expressing CHMO only under the same conditions.

Enhanced Biodegradation of Environmental Allergen by a vgb-containing Burkholderia cepacia

  • Kim, Mi-Sun;Yoon, Suk-Ran;Jun, Woo-Jin;Park, So-Young;Yang, Young;Shim, Sang-In;Hwang, Kwang-Woo;Chung, Jin-Woong
    • Molecular & Cellular Toxicology
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    • 제1권4호
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    • pp.262-267
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    • 2005
  • Using genetic engineering, the Vitreoscilla (bacterial) hemoglobin gene (vgb) was integrated stably into the chromosomes of and Burkholderia cepacia. Similar to previous results, the wild type VHb improved growth for Burkholderia cepacia and degradation of benzoic acid under both normal and low aeration conditions. The stable expression of VHb enhanced these parameters. The results demonstrate the possibility that the positive effects provided by VHb may be augmented by protein engineering.

In Vivo Wound Healing Activity of Crocodile (Crocodylus siamensis) Hemoglobin and Evaluation of Antibacterial and Antioxidant Properties of Hemoglobin and Hemoglobin Hydrolysate

  • Pakdeesuwan, Anawat;Araki, Tomohiro;Daduang, Sakda;Payoungkiattikun, Wisarut;Jangpromma, Nisachon;Klaynongsruang, Sompong
    • Journal of Microbiology and Biotechnology
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    • 제27권1호
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    • pp.26-35
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    • 2017
  • The hydrolysis of proteins constitutes an invaluable tool, granting access to a variety of peptide fragments with potentially interesting biological properties. Therefore, a hemoglobin (Hb) hydrolysate of Crocodylus siamensis was generated by digestion under acidic conditions. The antibacterial and antioxidant activities of the Hb hydrolysate were assessed in comparison with intact Hb. A disc diffusion assay revealed that the Hb hydrolysate exhibited antibacterial activity against eight strains of gram-positive bacteria and showed a higher efficacy than intact Hb. Moreover, the antioxidant activity of intact Hb and its hydrolysate was evaluated using ABTS and DPPH radical scavenging assays. The Hb hydrolysate exhibited free radical scavenging rates of 6-32%, whereas intact Hb showed a slightly higher activity. In addition, non-toxicity to human erythrocytes was observed after treatment with quantities of Hb hydrolysate up to $10{\mu}g$. Moreover, active fragmented Hb (P3) was obtained after purifying the Hb hydrolysate by reversed-phase HPLC. Scanning electron microscopy demonstrated the induction of bacterial cell membrane abnormalities after exposure to P3. Antibacterial and antioxidant activities play crucial roles for supporting the wound healing activity. Consequently, an in vivo mice excisional skin wound healing assay was carried out to investigate the effects of intact Hb treatment on wound healing in more detail. The results clearly demonstrate that intact Hb is capable of promoting 75% wound closure within 6 days. These findings imply that intact Hb of C. siamensis and its acid hydrolysate may serve as valuable precursors for food supplementary products benefitting human health.

Comparative Production of Green Fluorescent Protein Under Co-expression of Bacterial Hemoglobin in Escherichia coli W3110 Using Different Culture Scales

  • Bassapa Johnvesly;Kang, Dong-Gyun;Park, Suk-Soon;Kim, Ji-Hyun;Cha, Hyung-Joon
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제9권4호
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    • pp.274-277
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    • 2004
  • Production of green fluorescent protein (GFP) as a model foreign protein using different culture scales under co-expression of Vitreoscilla hemoglobin (VHb) in the industrial Escherichia coli strain W3110 (a K12 derivative), was examined. It was found that the VHb co-expressing W3110, exhibited an exceptional and sustained production ability during cell cultures using different scales, while the VHb non-expressing strain showed variable production levels. This high and sustained production ability indicates that the VHb co-expressing E. coli W3110, could be successfully employed for practical large-scale production cultures without the need for serious consideration of scale-up problems.

Cell growth and GFP expression in E. coli BL21 and W3110 under coexpression of Vitreoscilla hemogobin

  • 강동균;김연규;차형준
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2001년도 추계학술발표대회
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    • pp.754-757
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    • 2001
  • Expression of the vhb gene encoding bacterial hemoglobin (VHb) from Vitreoscilla has been used to improve recombinant cell growth and enhance product formation under microaerobic conditions because of its ability to enhance oxygen use. We coexpressed GFP and VHb in Escherichia coli BL21 and W3110, and compared with GFP control which was not expressed VHb. We used nar oxygen-dependent inducible promoter for VHb expression. The GFP amounts in E. coli expressed VHb was about five fold higher than in the control Fluorescence intensity was increased about two fold.

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천련자가 만성 비세균성 전립선염 Rat 모델에서 혈액 및 세포조직의 변화에 미치는 영향 (The Effects of Toosendan fructus(TOF) treatment on Hematological and Cyto-pathological Alterations in Non-Bacterial Prostatitis Rat Model)

  • 조충식;이구현;장선규;최정식;김철중
    • 대한본초학회지
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    • 제22권4호
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    • pp.145-153
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    • 2007
  • Objective : Although chronic non-bacterial prostatitis is a common disease, it is very difficult to treat effectively. Toosendan fructus(TOF) has been traditionally used in treatment of abdominal pain and parasite. In this study, we investigated the therapeutic effects and action mechanism of Toosendan fructus (TOF) in the rat model of non-bacterial prostatitis induced by castration and testosterone treatment. Method : Eight-month-old rats were treated with $17{\beta}$-estradiol after castration for induction of experimental non-bacterial prostatitis, which is similar to human chronic prostatitis in histophatological profiles. Toosendan fructus(TOF) and testosterone were administered as an experimental specimen and a positive control, respectively. The prostates were evaluated by histopahological parameters including the epithelial score and epithelio-stromal ratio for glandular damage. Also, the prostates were observed by hematological alterations of WBC, RBC, hemoglobin, hematocrit and platelet. Results : While prostates of control rats revealed severe acinar gland atrophy and stromal proliferation, the rats treated with Toosendan fructus(TOF) showed a diminished range of the tissue damage. Epithelial score was improved in Toosendan fructus(TOF) than that of the control. The epithelio-stromal ratio was lower in Toosendan fructus(TOF) when compared to that of the control. Also, the examination of bloods were not observed hematological change. Conclusions : These findings suggest that Toosendan fructus(TOF) may protect the glandular epithelial cells and may take hematological safety. We concluded that Toosendan fructus(TOF) may could be a useful remedy agents for treating the chronic non-bacterial prostatitis.

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인동소염방(忍冬消炎方)이 만성 비세균성 전립선염 Rat 모델에서 혈액 및 세포조직의 변화에 미치는 영향 (The Effects of IDS(Indongsoyeom-bang) Treatment on the Hematological and Cytopathological Alterations in Non-Bacterial Prostatitis Rat Model)

  • 이종빈;황석연;조충식
    • 혜화의학회지
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    • 제20권1호
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    • pp.91-104
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    • 2011
  • Background : Although chronic non-bacterial prostatitis is increasing, it is hard to treat effectively. In western medicine, antimicrobials drug, ${\alpha}$-adreno-ceptor antagonists, anti-inflammatory drugs, tricyclic antidepressants and anticholinergic agents are used commonly, but chronic prostatitis/chronic pelvic pain syndromes is confusing and frustrating for urologist. IDS(Indongsoyeom-bang) is used in treatment of chronic prostatitis/chronic pelvic pain syndromes. And it is reported that GLS(Gleditsiae spina) and TOF(Toosendan fructus) components of IDS have significant effect on protection of the glandular epithelial cells. Objective : In this study was conducted to investigate the therapeutic effects and action machanism of IDS in the rat model of non-bacterial prostatitis induced by castration and testosterone treatment. Methods : We observed six experimental objects of normal group, control group, testosterone group, and IDS 50 mg/kg, 200mg/kg, 400mg/kg group. Rats were treated with 17 ${\beta}$-estradiol after castration for induction of experimental non-bacterial prostatitis, which is similar to human chronic prostatitis in histophatological profiles. IDS and testosterone were administered as an experimental specimen and a positive control, respectively. The prostates were evaluated by histological parameters including the epithelial score and epithelio-stromal ratio for glandular damage. Also, the prostates were observed by Hematological alterations of WBC, RBC, hemoglobin and platelet. Results : While prostates of control rats revealed severe acinar gland atrophy and stromal proliferation, the rats treated with IDS-50 showed a diminished range of the tissue damage. Epithelial score was improved in IDS than that of the control. The epithelio-stromal ratio was lower in IDS when compared to that of the control. Also, the examination of bloods were not observed hematological change. Conclusion : These finding suggests that IDS may protects the glandular epithelial cells. We concluded that IDS could be a useful remedy agent for treating chronic non-bacterial prostatitis.

High-Efficiency Generation of Monoclonal Antibody for Vitreoscilla Hemoglobin Protein

  • Kim, Eun-Mi;Kim, Myung-Hee;Kim, Min-Gon;Kim, Sang-Woo;Ro, Hyeon-Su
    • Journal of Microbiology and Biotechnology
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    • 제22권2호
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    • pp.226-229
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    • 2012
  • Bacterial hemoglobin from Vitreoscilla (VHb) is recognized as a good fusion protein for the soluble expression of foreign protein. In this study, we generated a monoclonal antibody (MAb) against VHb for its detection. For the rapid screening of MAb, a protein chip technology based on the Alexa-488 (A488) dye labeling method was introduced. In order to fabricate the chip, the VHb protein was chemically coupled to the chip surface and then the culture supernatants of 84 hybridoma cell lines were spotted onto the VHb chip. The bound MAbs were measured by A488-modified anti-mouse IgG. A single spot (MAb A10) exhibited significantly high signal intensity. The immunoblot analysis evidenced that the MAb A10 can detect VHb-fused proteins with high specificity.