• Title/Summary/Keyword: and biotin

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Gene Cloning, Expression, and Characterization of a New Carboxylesterase from Serratia sp. SES-01: Comparison with Escherichia coli BioHe Enzyme

  • Kwon, Min-A;Kim, Hyun-Suk;Oh, Joon-Young;Song, Bong-Keun;Song, Jae-Kwang
    • Journal of Microbiology and Biotechnology
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    • v.19 no.2
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    • pp.147-154
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    • 2009
  • The carboxylesterase-encoding gene(bioHs) of a newly isolated strain, Serratia sp. SES-01, was cloned from the genomic DNA library by detecting formation of transparent halo around the colony on LB-tributyrin agar plates. The amino acid sequence of BioHs was highly similar to the members of the BioH enzyme family involved in the biotin biosynthetic pathway; it showed the highest similarity(91%) with that of Serratia proteamaculans. To compare BioHs with other BioH enzymes, the relatively well-known bioHe gene of E. coli was cloned with PCR. After we achieved high-level expression of soluble BioHs and BioHe through the exploration of different culture conditions, the purified BioHs and BioHe enzymes were characterized in terms of specificity, activity, and stability. BioHe was generally more robust to a change in temperature and pH and an addition of organic solvents than BioHs. The two enzymes exhibited a strong preference for carboxylesterase rather than for thioesterase and were optimal at relatively low temperatures($20-40^{\circ}C$) and alkaline pHs(7.5-9.0). The results in this study strongly suggested that both the BioHs and BioHe enzymes would be potential candidates for use as a carboxylesterase in many industrial applications.

Identification of the Antibiotic-Producing Chstridium sp. KH-431 and Purification of the Antibiotics (항생물질을 생산하는 Clostridium sp. KH-431의 동정 및 항생물질의 정제)

  • 홍수형;김미정;박용복;이재근;하지홍
    • Microbiology and Biotechnology Letters
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    • v.21 no.1
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    • pp.41-46
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    • 1993
  • A strain showing antibiotic activities against various bacteria and fungi was selected from approximately 2,000 microorganisms obtained from soil samples. This strain, designated as KH-431, was identified as a Clostridium sp. by its morphological, physiological and biochemical characteristics. The highest production of the antibiotics was achieved in a fermentation medium containing sorbitol, yeast extract, d-biotin and $CaCl_2$ The antibiotics were isolated from the culture broth by solvent extraction using ethyl acetate, silica gel column chromatography and recrystallization. Two kinds of antibiotics, KG-431A and KG-431B were obtained after the purification procedure, and only KG-431B was successful to recrystallize.

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Studies on the Fermentative Utilization of Cellulosic Wastes (Part 8) Mixed Culture of Cellulose Assimilating Bacteria (폐섬유자원의 발효공학적 이용에 관한 연구 (제8보) 섬유소자화세균의 혼합배양)

  • 윤한대;성낙계
    • Microbiology and Biotechnology Letters
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    • v.6 no.2
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    • pp.51-57
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    • 1978
  • The study was made of the cultural condition and physiological characteristics of the symbiotic pair of microorganisms, Cellulomonas flavigena and the second organism. It also contains the results of a taxonomical study of the second organism. The results obtained wers summarized as follows : 1) Cell yield of the mixed culture, Cellulomonas and the second organism, was higher than that of each pure culture in CM-Cellulose medium. 2) The taxonomical characteristics of the second organism revealed that it probably belonged to the genus Sporocytophaga because it had a gliding motility and microcyst. 3) Optimum pH of the mixed culture was found to be in the vicinity of 7.2, and optimum temperature of the cell growth in the mixed culture was observed to be in the vicinity of 30$^{\circ}C$. 4) It was found that the majority of the population during growth in the mixed culture consisted of Cellulomonas flavigena. 5) Cellulomonas flavigena required thiamine and biotin as growth factors but Sporocytophaga sp. had no requirement of vitamins. 6) Gulucose was not found in detectable amounts in the medium of Cellulomonas flavigena but it was traced in the mixture by thin layer chromatography. 7) Sixteen amino acids were analyzed from the cell protein of Cellulomonas flavigena by amino acid autoanalyzer. The amount of the leucine, valine and arginine was very high.

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Studies on the Penicillinase Produced by a Streptomyces sp. (Part I). Optimal Conditions for the Penicillinase Production by Streptomyces sp. YS-40. (Streptomyces sp.가 생산하는 Penicillinase 에 관한 연구 (제1보) Streptomyces sp. YS-40에 의한 Penicillinase의 생산조건)

  • 도재호;김상달;이동의
    • Microbiology and Biotechnology Letters
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    • v.10 no.3
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    • pp.177-184
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    • 1982
  • Studies were carried out to investigate the optimal culture conditions for the production of penicillinase using a strain of Streptomyces sp. isolated from soil, YS-40. Among the carbon and nitrogen sources, glucose and L-asparagine increased the peniciilinase production. The addition of M $n^{++}$, $Ca^{++}$ and L $i^{+}$ increased the enzyme production, but depressed by F $e^{+++}$, F $e^{++}$, $Mg^{++}$, Z $n^{++}$, A $g^{++}$, $Ba^{++}$ and S $n^{++}$. L-Leucine slightly increased the enzyme production but L-histidine, L-methionine depressed. Among the vitamins riboflavine, i-inositol, hesperidine, niacin-amide, biotin, folic acid, DL-$\alpha$-lipoic acid increased the enzyme formation. The addition of cephradine, cephalexin, ampicillin, cloxacillin more increased the enzyme formation than that of other$\beta$-lactam antibiotics and antibiotics. Optimal pH and temperature on the enzyme formation was pH 7.0 and 28$^{\circ}C$ respectively Amount of the enzyme production reached at maximum with incubation for 3 days on the optimal condition.

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Studies on the L-Glutamic acid Fermentation(Part I ) L-Glutamic acid Production from the Hydrolyzate of Sliced and Dried Sweet Potatoes (L-Gutamic acid희효생산에 관한 연구(제일보) 절간고구마 산분해액을 이용한 L-Glutamic acid 생산)

  • 양한철;최용진;양한우;성하진
    • Microbiology and Biotechnology Letters
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    • v.3 no.1
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    • pp.7-15
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    • 1975
  • The possibilities of utilizing acid-hydrolyzate of "Sliced and dried sweet potatoes" as a carbon source for the microbial production of L-Glutamic Acid(L-GA) with Micrococcus glutamicus were investigated and the results showed as follows: 1) The highest hydrolysis rate, 74.6% of the reducing sugar based on the weight of dry matter, was obtained when the sweet potatoes were hydrolyzed with 0.8% of HCI at 2.0kg/$cm^2$ for 30 minutes. The most favorable hydrolyzate for the growth of the cells, however, was found to be the one obtained by treating the sweet potatoes with 0.5% HCI at 2. 0kg/$cm^2$ for 10 minutes. Reducing sugar content of the hydrolyzate was 10% as glucose. 2) Biotin content of the hydrolyzate was 25$\mug$/1 and it was proved to be excess in amount for the L-GA production. 3) The effects of addition of antibiotics, alcohols and fatty acid esters on the L-GA production were tested in the biotin excess medium. The production of L-GA was most increased to 32.5g/l with the addition of 10 I. U. of penicillin per ml. to the culture medium at 4 hours after inoculation. But the addition of alcohols, especially fatty acid esters, showed no significant effects. 4) Among the organic nutrients tested. " Gluten acid hydrolyzate" greatly enhanced the production of L-GA adding it's concentration of 1.0% to the medium. 5) The maximum production of L-GA resulted in 35g/1 when the cells were grown for 48 hours in the hydrolyzate medium supplemented with 1.0% of "Gluten acid hydrolyfate" and with 10 I. U. of penicillin per ml added at 4 hours after cultivation.

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Production Conditions of Two Polysaccharides from Marine Bacterium Zoogloea sp. (해양세균 Zoogloea sp.로부터 두 종류의 polysaccharide 생산조건)

  • 장명웅;강양순
    • KSBB Journal
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    • v.10 no.5
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    • pp.518-524
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    • 1995
  • Marine bacterium, as a microbial source producing polysaccharides, was newly isolated from the eastern and western sea of Korea and was identified as Zoogloea sp. (KCCM 10036). It produced two different types of polysaccharides, especially: WSP (water-soluble polysaccharide) and CBP (cell-bound polysaccharide). The former was isolated from the supernatant of centrifuged broth by acetone precipitation, and the latter was isolated from the pellet by acetone and CPC (cetylpyrldinium chloride) precipitation. The productivity of polysaccharides were increased with the addition of promoting agents such as biotin, ampicillin and surfactant. After batch fermenting, the productivity of WSP and CBP were reached to maximum values of $9.Og/\ell$, $2.5g/\ell$ in the culture medium containing 1% of glucose as a carbon source.

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Identification of surface antigen of Trichomonas vaginalis (질편모충의 표면항원 분석)

  • 민득영;임미혜
    • Parasites, Hosts and Diseases
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    • v.32 no.4
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    • pp.243-248
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    • 1994
  • Plasma membrane proteins of a Korean isolate of Trichomonus vofinalis HY-1 were fractionated for antigen analysis. Homogenates of T. vaginalis were fractionated by the differential centrifugation using sucrose step-gradient method. The interface layer from the 25%/45% sucrose was collected as a plasma membrane fraction and its purity was examined by transmission electron microscopy. The antigenicity of plasma membrane fraction was analysed by enzyme-linked immunoelectrotransfer blot technique with immune rabbit serum and compared with surface antigen labelled with N. hydroxysuccinimide-biotin. The fluffy fraction of 25%/45% sucrose interface was homogeneous and membrane particles were present as extended sheet and concentric vesicles showing typical trilamellar appearance under transmission electron microscope. Seven fractions at 40, 50, 60, 110, 130, 140 and 150 kDa were identified as the antigenic membrane proteins in EITB with anti HY-1 rabbit serum. The common band at 60 kDa was detected both in antigenic fractions of plasma membrane and surface protein labelled with NHS-biotin. This result indicates that this protein is considered as a major surface antigen of T. vaginalis. The role of this surface antigen at 60 kDa should be studied further.

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Effects of Vitamins on the Differentiation of Preadipocytes from Hanwoo Cattle Adipose Tissues

  • Lee, H.J.;Lee, S.C.;Oh, Y.K.;Han, In K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.4
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    • pp.446-450
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    • 2000
  • The experiment was conducted to study the effects of water soluble vitamins and retinoic acid on the differentiation of preadipocyte from omental, subcutaneous, intermuscular and intramuscular adipose tissue of Hanwoo. Differentiation was assessed by the change in enzyme activity, glycerol-3 phosphate dehydrogenase in serum free cell culture system. Preadipocytes treated with biotin ($10{\mu}M$) and pantothenic acid ($100{\mu}M$) were significantly (p<0.05) less differentiated than those from the control in all adipose tissue depots except intramuscular tissue. Although there was no significance, vitamin C was shown to stimulate the adipocyte conversion in omental and subcutaneous, but not in intermuscular and intramuscular adipose tissues. Lower values of GPDH activity in intermuscular preadipocyte were interpreted to be caused by relatively higher amounts of protein. In this experiment vitamin C did not stimulate fat deposition in intramuscular adipose tissue but further experiments are needed on the role of vitamin C in preadipocyte differentiation. When treated with different levels of retinoic acids, differentiation of preadipocytes was significantly (p<0.05) reduced from the level of $0.5{\mu}g/ml$ in omental and intermuscular, from $50{\mu}g/ml$ in subcutaneous, and in intramuscular at $500{\mu}g/ml$, thus showing that intramuscular preadipocytes were least responsive to retinoic acid in differentiation. All-trans retinoic acid appeared to inhibit the differentiation in a dose dependent manner, regardless of adipose tissues type.

Studies on a methanol-assimilating yeast for the production of Single Cell Proein (미생물(微生物) 단백질(蛋白質)을 생산(生産)하기 위(爲)한 메탄올 자화효모(자화효모)에 관(關)한 연구(硏究))

  • Chung, Hee-Jong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.15 no.4
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    • pp.24-31
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    • 1986
  • A methanol-assimilating yeast for the production of Single Cell Protein was isolated from the soil and identified. Methanol as a sole carbon source was used in this study. The strain was identified as Candida boidinii which grew best at the initial concentration of methanol at 2% , with the addition of 0.5% methanol at every 12 hours, The Single Cell Protein production was maximal after 72 hours of incubation at pH 5.0, $30^{\circ}C$. Thiamin and biotin were stimulated the growth of this yeast at the levels of $1000{\mu}g/{\ell}$ and $10{\mu}g/{\ell}$respectively.

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Specific Binding of Streptavidin onto the Nonbiofouling Titanium/Titanium Oxide Surface through Surface-Initiated, Atom Transfer Radical Polymerization and Bioconjugation of Biotin

  • Kang, Sung-Min;Lee, Bong-Soo;Kim, Wan-Joong;Choi, In-Sung S.;Kil, Mun-Jae;Jung, Hyuk-Jun;Oh, Eu-Gene
    • Macromolecular Research
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    • v.17 no.3
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    • pp.174-180
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    • 2009
  • Chemical modification of titanium/titanium oxide (Ti/$TiO_2$) substrates has recently gained a great deal of attention because of the applications of Ti/$TiO_2$-based materials to biomedical areas. The reported modification methods generally involve passive coating of Ti/$TiO_2$ substrates with protein-resistant materials, and poly(ethylene glycol) (PEG) has proven advantageous for bestowing a nonbiofouling property on the surface of Ti/$TiO_2$. However, the wider applications of Ti/$TiO_2$ based materials to biomedical areas will require the introduction of biologically active moieties onto Ti/$TiO_2$, in addition to nonbiofouling property. In this work, we therefore utilized surface-initiated polymerization to coat the Ti/$TiO_2$ substrates with polymers presenting the nonbiofouling PEG moiety and subsequently conjugated biologically active compounds to the PEG-presenting, polymeric films. Specifically, a Ti/$TiO_2$ surface was chemically modified to present an initiator for atom transfer radical polymerization, and poly(ethylene glycol) methacrylate (pEGMA) was polymerized from the surface. After activation of hydroxyl groups of poly(pEGMA) (pPEGMA) with N,N'-disuccinimidyl carbonate, biotin, a model compound, was conjugated to the pPEGMA films. The reactions were confirmed by infrared spectroscopy, X-ray photoelectron spectroscopy, contact angle goniometry, and ellipsometry. The biospecific binding of target proteins was also utilized to generate micropatterns of proteins on the Ti/$TiO_2$ surface.