• Title/Summary/Keyword: Intermittent Feeding

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The Production of Sodium Gluconate by Aspergillus niger (Aspergillus niger를 이용한 글루콘산 나트륨의 생산)

  • 이현철;정봉우
    • KSBB Journal
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    • v.11 no.1
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    • pp.65-70
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    • 1996
  • Sodium gluconate was produced by neutralization of gluconic acid formed during the submerged culture fermentation of glucose with Aspergillus niger ACM 7. The fermentation characteristics of Aspergillus niger ACM 7 were investigated quantitatively according to the change of the initial glucose concentrations and the initial pHs of fermentation broth. The maximum specific growth rate was estimated to be $0.20hr^{-1}$ at 95g/$\ell$ of initial glucose concentration. The maximum fermentability of sodium gluconate was 95% at the initial glucose concentration of 26g/$\ell$. However, the maximum sodium gluconate productivity was 1.18g/$\ell$/hr when the initial glucose concentration was 110g/$\ell$. The optimum pH was found to be 5.5 for both the cell growth and the sodium gluconate production. With optimized culture conditions, the productivity of sodium gluconate in a fed-batch culture(production fermentor, 16,000$\ell$) increased up to 7.1g/$\ell$/hr.

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Characterization of the ${\beta}-Cyclodextrin$ Glucanotransferase Bacillus firmus var. alkalophilus and Its Expression in E. coli

  • Park, Tae-Hyung;Shin, Hyun-Dong;Lee, Yong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.811-819
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    • 1999
  • The ${\beta}-CGTase$ gene of alkalophilic Bacillus firmus var. alkalophilus was cloned into E. coli using $pZErO^{TM}-2$ as a vector. The cloned gene encoded a total of 710 amino acid residues consisting of 674 amino acids of the matured protein and 36 amino acids of the signal peptide, including 20 amino acids from the lacZ gene in the vector. Although the cloned ${\beta}-CGTase$ gene did not contain the promoter and start codons, it was expressed by the lac promoter and lacZ start codon in the $pZErO^{TM}$ vector. A comparison was made with the amino acid sequence and ten other CGTases from Bacillus sp. Also, ten highly conserved regions, which are important amino acid residues in catalysis of CGTase, were identified. The lac promoter used for expression of the ${\beta}-CGTase$ gene was induced constitutively in recombinant E. coli even without IPTG possibly because of a lack of the lacI gene in both host and vector, repressing the lacZ gene in the lac operon. Its expression was catabolically repressed by glucose, however, its repression was reduced by soluble starch, mainly because of the extremely high increase of the cAMP level. ${\beta}-CGTase$ can be overproduced in the recombinant E. coli by maintaining intracellular cAMP levels mostly through the intermittent feeding of glucose during cultivation.

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Poly-$\beta$-Hydroxybutyrate Produced by Pink-Pigmented Facultative Methylotrophic Bacterium from Methanol (분홍색 통성 메탄올 자화세균이 생산하는 Poly-$\beta$-Hydroxybutyrate)

  • 송미연;이재호;이용현
    • Microbiology and Biotechnology Letters
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    • v.18 no.3
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    • pp.273-279
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    • 1990
  • For poly- $\beta$ -hydroxybutyrate (PHB) production, a pink-pigmented facultative methylotrophic bacterium (PPFM) P-10 was newly isolated from soils through methanol-enrichment culture. The optimal medium composition for cell growth was 1.0% (vlv) of methanol as carbon source and l.Og/l of ,TEX>$NH_4Cl$, equivalent to C/N ratio of 13.2 at pH 7.0 and $30^{\circ}C$. To investigate the optimal condition for YHB accumulation, two-stage culture technique was adopted; first stage for cell growth and second stage for accumulation of PHB providing unbalanced growth conditions. The optimal PHB accumulation was 1.0% (vIv) of methanol and 0.26gll of $NH_4Cl$, C/N of 50.8 at pH 6.0. To overcome methanol inhibition on cell growth, intermittent feeding fed-batch culture technique was employed, and the cell concentration as high as 14gll with 40% of PHB was achieved. The purified PHB was identified using IR and $1^H NMR$ as homopolymer of 8hydroxybutyric acid. The absorption spectrum of extracted pink colored microbial pigment was alsa investigated.

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Optimization for Lacticin SA72 Production by Lactococcus lactis SA72 Isolated from Jeot-gal. (젓갈에서 분리한 Lactococcus lactis SA72에 의한 Lacticin SA72의 생산 최적화)

  • 백현동;구경모;김진곤;이나경
    • Microbiology and Biotechnology Letters
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    • v.31 no.1
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    • pp.46-50
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    • 2003
  • Lactococcus lactis SA72 from Jeot-gal (Korean traditional fermented fish foods) produces lacticin SA72. The influence of several parameters on the fermentative production of lacticin SA72 by Lactococcus lactis SA72 was studied. MRS medium among several media was selected for enhanced bacteriocin production. The mean growth rate and bacteriocin productivity of L. lactis SA72 increased as the initial pH of the media increases. The highest lacticin SA72 activity was detected 3,200 AU/ml at pH 6.0, $32^{\circ}C$, and 1% (inoculum size, v/v) in the jar fermenter. Enhanced production of lacticin SA72 was investigated by a fed-batch cultivation with the intermittent feeding of the concentrated glucose solution. Under the optimized conditions, lacticin SA72 activity finally reached to 6,400 AU/ml.

Field evaluation of Enterotoxigenic Escherichia coli-specific bacteriophage (ΦCJ19) as a feed additive (Enterotoxigenic Escherichia coli 특이 박테리오파지 (ΦCJ19)의 사료 첨가제로서의 사양 평가)

  • Yoo, Anna;Cha, Seung Bin;Shin, Min Kyoung;Park, Hong Tae;Seo, Hyo Sil;Kim, Jae Won;Yoo, Han Sang
    • Korean Journal of Veterinary Research
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    • v.53 no.2
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    • pp.83-88
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    • 2013
  • Field efficacy of enterotoxigenic Escherichia coli-specific phage (${\Phi}CJ19$) as a feed additive was evaluated in weaning piglets. Fifty-four piglets at 3-4 weeks old were allocated in three different groups and two of them were fed with bacteriophage at different concentrations ($10^6PFU/kg$ feed and $10^8PFU/kg$ feed, respectively) for 30 days. Body weight and feed intake were measured at 10 days interval and body condition and fecal score were inspected every day. Based on the measurement, feed conversion rate (FCR) and average daily gain (ADG) of each group during 30 days were analyzed. The analysis suggests that the bacteriophage may help the improvement of FCR and ADG at $10^8PFU/kg$ of bacteriophage feeding group in 30 days. A result from analysis of fecal score indicates that the bacteriophage also may help to relieve the intermittent diarrhea in post-weaning stage. Those results suggest that bacteriophage might help the growth of piglets in post-weaning stage.

MANDIBULAR DISTRACTION OSTEOGENESIS IN AN INFANT WITH PIERRE ROBIN SEQUENCE: REPORT OF A CASE (Pierre Robin sequence 환아에서 하악골신장술)

  • Ryu, Sun-Youl;Kwon, Jun-Kyong;Kim, Sun-Kook
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.34 no.4
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    • pp.460-467
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    • 2008
  • Pierre Robin sequence as a symptom triad of micrognathia, glossoptosis, and cleft palate results in upper airway obstruction and feeding problems. If mild, it is often managed in the prone position. When positional treatment fails, however, surgical intervention such as tongue-lip adhesion, tracheostomy, and mandibular distraction osteogenesis is mandatory to relieve airway obstruction. There has been growing interest in the application of distraction osteogenesis for the management of craniofacial abnormalities. The mandibular distraction osteogenesis to newborns may prevent the airway obstruction, decrease the potential tracheostomy, and reduce the likehood of orthognathic surgery after growth. We experienced an infant with Pierre Robin sequence who showed mandibular hypoplasia, glossoptosis, incomplete cleft palate, intermittent cyanos is, depression of the chest, and respiratory difficulty associated with airway obstruction. We treated the airway obstruction by tongue-lip adhesion at 2 weeks of age, and treated the mandibular retrognathism and depression of the chest byusing internal mandibular distraction osteogenesis at 7 month of age. The mandible moved forwardly, the upper airway space was enlarged, and the antero-posterior distance of the mandible was elongated after the mandibular distraction. Mandibular distraction osteogenesis may be a promising technique to avoid the need of tracheostomy and orthognathic surgery, and to correct airway obstruction in infants with congenital craniofacial malformation.

Simultaneous Overpexpression of Genes Encoding Cellulose- and Xylan-Degrading Enzymes through High Density Culture of a Recombinant Yeast Cell (재조합 효모 세포의 고농도배양을 통한 섬유소와 자일란 분해효소 유전자의 동시 과발현)

  • Kim, Yeon-Hee;Heo, Sun-Yeon;Kim, Gun-Do;Nam, Soo-Wan
    • Microbiology and Biotechnology Letters
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    • v.46 no.4
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    • pp.390-394
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    • 2018
  • For the coexpression of endoxylanase and endoglucanase genes in yeast Saccharomyces cerevisiae, the genes were separately inserted downstream of the yeast ADH1 promoters, resulting the plasmid pAGX3 (9.83 kb). In the batch culture on YPD medium of the yeast transformant, S. cerevisiae SEY2102/pAGX3, the total activities of the enzymes reached about 7.91 units/ml for endoxylanase and 0.43 units/ml for endoglucanase. In the fed-batch culture with intermittent feeding of yeast extract and glucose, the total activities of 24.9 units/ml for endoxylanase and 0.84 units/ml for endoglucanase were produced which were about 3.1-fold and 2.0-fold increased levels, respectively, compared to those of the batch culture. Most of endoxylanase and endoglucanase activities were found in the extracellular media. This recombinant yeast could be useful for the development of simultaneous saccharification bioprocess of the cellulose and xylan mixture.

Sewage Treatment Using a Double Media Reed Constructed Wetland (복층여재 갈대 인공습지에 의한 생활하수 처리)

  • Seo, Jeoung-Yoon
    • Journal of Wetlands Research
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    • v.16 no.4
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    • pp.379-388
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    • 2014
  • A sewage was treated using a serially combined vertical(VFCW) and horizontal flow double media (sand and zeolite for VFCW and sand and waste oyster-shell for HFCW) reed constructed wetland(HFCW) with intermittent feeding (see Fig. 1). The sewage was fed into the reed constructed wetland for 10 minutes every 6 hours at the hydraulic load of $314L/m^2{\cdot}day$. The summarized results were as follows: pH values in the effluent depended very heavily on oyster-shell height filled in the HFCW. They were maintained at less than pH 6.24 when the height of the oyster-shell layer was 200 mm. Influent DO(oxygen demand) values(average 0.19 mg/L) were increased in the VFCW(average 7.65 mg/L) and decreased again in the HFCW(average 6.49 mg/L). They were higher in the winter than in the summer. The OTR(oxygen transfer rate) was $57.15g\;O_2/m^2{\cdot}day$ in the VFCW and $5.65g\;O_2/m^2{\cdot}day$ in the HFCW. The removal efficiency of $NH_4{^+}$-N was 80.17%(6.01 $NH_4{^+}$-N mg/L in the effluent). It was lower than that in the case where only zeolite was filled in the reed constructed wetland. But it was expected that treated sewage effluent using a double media reed constructed wetland with 300 mm zeolite layer could stably meet the Korean treated sewage effluent standard(20 mg T-N/L). Average removal efficiencies were SS 88.09%, BOD 88.12%, $COD_{Cr}$ 83.11%, $COD_{Mn}$ 85.58%, T-N 57.21%, $NH_4{^+}$-N 80.17%, T-P 86.73%. Nearly, The concentration of $NO_3{^-}$-N in the effluent of the VFCW was decreased in that of the HFCW. More than half of T-N in the effluent was $NO_3{^-}$-N(7.92 mg/L) but the concentration of $NO_2{^-}$-N in the effluent was average 0.90 mg/L. The removal efficiencies of T-P were 93.24%, 86.30% and 55.44% at the height of the oyster-shell-filled constructed wetland of 800 mm, 500 mm and 200 mm, respectively and therefore, they were proportional to oyster-shell height filled in the HFCW.

Selection of Constitutive Promoter for Exoinulinase Production in Fed-Batch Culture of Recombinant Yeast (재조합 효모의 유가배양에서 Exoinulinase생산을 위한 Promoter의 선별)

  • 김이경;고지현;김연희;김성구;남수완
    • Microbiology and Biotechnology Letters
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    • v.29 no.4
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    • pp.206-211
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    • 2001
  • In order to overexpress constitutively the Kluyveromyces marxianus exoinulinase gene (INUI) in Saccharomyces cerevisiae, four episomal expression systems employing GAPDH, ADHI, PGK and ENOI promoters were constructed as p YIGP aADHI -INU, pPGK-INU, and pENOI- INU plasmids respectively, When S cereviais transformants harboring each plasmid were batchwisely cultivated in the fermentor containing 5% glucose medium no significant differences in the cell growth are observed How- ever the experession level of exoinulinase and plasmid stability showed a strong dependency on the promoter employed. The expression levels of exoinulinase were about 1.70 unit/ml for GAPDH promoter 1.67 unit/ml for PGK promoter, 1.29 unit /ml for ADH1 promoter, and 0.80 unit/ml for ENOl promoter. The plasmid stabilites were maintaines above 80% in all experession systems. except the GAPDH promoter system of 55%, Based on the plas- mid stability and expression level of exoinulinase the ADHl and PGK promoter system were selected for the fed - batch culture to overproduce exoinulinase By the intermittent feeding of yeast extract and glucose, both promoter systems gave the cell concentration of about 30 g-dry cell weight/1 byt the maximal exoinulinase activity of 3.70 unit/ml and plasmid stability of 96% in the ADH1 promoter were higher than those (2.70 unit/ml, 80%) of PGK sys- tem Taking into account the plasmid stability and extended culture time the ADH1 promoter systems would be the most feasible expression systems for the constitutive overproduction of exoinulinase through high cell-density fed- batch cultures using non-selective rich medium.

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Accumulation of Poly-$\beta$-Hydroxybutyrie Acid by Alcaligenes sp. (Alcaligenes sp.에 의한 Poly-$\beta$-Hydroxybutyric Acid의 축적)

  • 임명순;손홍주;박수민;이종근;이상준
    • Microbiology and Biotechnology Letters
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    • v.20 no.4
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    • pp.363-370
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    • 1992
  • Microorganisms capable of accumulating poly-p-hydroxybutyric acid(PHB) were isolated from soil by enrichment culture technique. Among them, the strain designated as FL-027 had high PHB productivity and was identified as Alcaligenes. The optimal medium composition for cell growth was 8.0 $g/\ell$ of fructose and 3.0 $g/\ell$ of $(NH_4)_2S0_4$, equivalent to C/N ratio 5.04 at pH 7.0 and $30^{\circ}C$. To investigate the optimal conditions for the PHB accumulation, we divided the process into two stages; the first stage for the growth of the cell in nutrient-rich medium and the second stage for the PHB accumulation in nutrient-deficiency medium. The optimal conditions for PHB accumulation were 8.0 $g/\ell$ of fructose and 0.25 $g/\ell$ of $(NH_4)_2S0_4$, equivalent to C/N ratio 60 at pH 6.5 and $30^{\circ}C$. PHB accumulation was stimulated by deficiency of nutrients such as $NH_4^+$, $Ca^{2+}$, $SO_4^{2+}$ in medium. Among them. $NH_4^+$ deficiency was chosen because of its effectiveness. We found the inhibition of cell growth by fructose in batch culture. In order to keep the fructose concentration at an optimal level, intermittent feeding fed-batch culture was employed, and the cell concentration as high as 10.83 $g/\ell$ whose PHB content was responsible for 43% of the dry cell weight. The purified PHB was identified as homopolymer of 3-hydroxybutyric acid by using IR and $^1H-NMR$.

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