• Journal of Microbiology and Biotechnology
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• 제3권1호
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• pp.39-45
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• 1993

The effect of concentrations of phosphate buffer and acrylonitrile, pH, and various salts on the operational stability of the immobilized cells of Brevibacterium CH2 in a packed bed reactor were investigated. The effects of salts and carriers on the swelling of the immobilized beads during hydrolysis in a columnreactor were also investigated. Immobilization of the cells in Ba-alginate was more desirable than those in polyacrylamide and Ca-alinate for the swelling of the immobilized beads and the desired quality of the acrylamide produced. High quality acrylamide was produced using the Ba-alginate beads in a recycle fed-batch reactor without using an isotonic substrate. The conversion yield was nearly 100％, including a trace amount of acrylic acid produced as a by-product.

• Journal of Microbiology and Biotechnology
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• 제9권4호
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• pp.482-487
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• 1999

The Pseudomonas sp. strain NGK1 (NCIM 5120) was immobilized in calcium alginate, agar, and polyacrylamide gel matrices. The salicylic acid-producing capacity of freely suspended cells was compared with immobilized cells in batches with a shake culture and continuous culture system in a packed bed reactor. Freely suspended cells ($4\times10^{10}cfu/ml$) produced 12 mM of salicylic acid, whereas cells immobilized in calcium alginate ($1.8\times10^{11}$cfu/g beads), agar ($1.8\times10^{11}$cfu/g beads), and polyacrylamide ($1.6\times10^{11}$cfu/g beads) produced 15, 11, and 16mM of salicylic acid, respectively, from naphthalene at an initial concentration of 25 mM. The continuous production of salicylic acid from naphthalene was investigated in a continuous packed bed reactor with two different cell populations. The longevity of the salicylic acid-producing activity of the immobilized cells from naphthalene was also studied in semi continuous fermentations. The immobilized cells could be reused 18, 13, and more than 20 times without losing salicylic acid-producing activity in calcium alginate-,agar-, and polyacrylamide-entrapped cells, respectively. The study reveals a more efficient utilization of naphthalene and salicylic acid production by the immobilized Pseudomonas sp. strain NGK1 as compared to the free cells.

• Journal of Microbiology and Biotechnology
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• 제7권4호
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• pp.250-257
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• 1997

A new sensing system based on the immobilization of luminescent bacteria, photobacterium phosphoreum, was proposed for continuous real-time monitoring of pollutants. The response curves demonstrate that Photobacterium phosphoreum immobilized on the strontium alginate were very sensitive to seven reference chemicals used. The significant inhibitory concentrations for bioluminescence emission were 5 ppm for Pb$(NO_3)_2$), $NiCl_2$, $CdCl_2$, 50 ppm for $NaASO_2$, 0.1 ppm for $HgCl_2$, 0.5 ppm for pentachlorophenol and less than 5 ppm for SDS, respectively. The alginate mixed-cells (AMC) retained their luminescence during experimental period (29 days) under storage condition of $-80^{\circ}C$. The variables affecting performance of continuous flow through monitoring (CFTM) was optimized in order to ensure stability and efficiency. The flow through cell with strontium-alginate immobilized luminescent bacteria was tested with salicylate and 4-nitrophenol. A rapid response of luminescence was recorded by time drive mode in bioluminescence spectrometer after exposure to both toxicants.

• Journal of Microbiology and Biotechnology
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• 제7권1호
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• pp.62-67
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• 1997

To enhance the hyperproductive and low energy-consuming ethanol fermentation rate, the thermotolerant yeast S. cerevisiae RA-74-2 cells were immobilized. An efficient immobilization condition was proved to be $1.5{\%}$ (w/v) alginate solution, neutral pH and 20 h activation of beads. The fermentation characteristics and stability at various temperatures were examined as compared with free S. cerevisiae RA-74-2 cells. The immobilized cells had excellent fermentation rate at the range of pH 3-7 at 30-$42^{\circ}C$ in 15-$20{\%}$ glucose media. When the seed volume was adjusted to 0.12 (v/v) (6ml bead/50 ml medium), $11{\%}$ (w/v) ethanol was produced during the first 34 hand $12.15{\%}$ (w/v) ethanol [$95{\%}$ (w/v) of theoretical yield] during the first 60 h in $25{\%}$ glucose medium. In repetitive fermentation using a 2 litre fermentor, 5.79-$7.27{\%}$ (w/v) ethanol [76-$95{\%}$ (w/v) of theoretical yield] was produced during the 40-55 h in $15{\%}$ glucose media. These data suggested the fact that alginate beads of thermotolerant S. cerevisiae RA-74-2 cells would contribute to economic and hyperproductive ethanol fermentation at high temperature.

• KSBB Journal
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• 제11권1호
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• pp.77-85
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• 1996

에탄올 생산 효모균주인 Saccharomyces K35의 에탄올 생산을 위한 최적의 초기 pH는 5.0으로 나 타났으며, 기질로 200g/$\ell$ 의 glucose를 이 용했을때 약 80%의 수율을 나타내어 내당성도 좋은 것으로 나타났다. 첨가제와 가교제로서 Celite R-634 1.67 %(v/v) 와 glutaraldehyde 1.67% (v/v) 를 함께 첨가했을때(ACG bead) Ca-alginate bead에 비해 안정성, 에탄올 생산성 빛 세포생존률이 좋게 나타 났다. 또한 배지밖에서 자란 효모 농도도 감소하여 A A CG bead가 Ca-alginate bead보다 phosphate이 온에 더 안정한 것으로 보인다. SEM (Scanning E Electron Microscopy)을 통해 ACG gel bead의 구조를 관찰하였는데 , ACG bead가 Ca~alginate bead에 비해 훨씬 온전한 모습을 유지하며, 효모의 농도가 상당히 밀집되어 있음을 보여준다. 반복회분 식 배양을 시도했을때 Ca-alginate bead의 경우는 7회분(약 40일)까지는 에탄올 놓도와 효모농도가 각각 138g/$\ell$-gel와 29~30g/$\ell$-gel을 유지하다가 급격히 감소하는 경향을 보였으나, ACG bead의 경 우에는 130~150g/$\ell$-gel와 32~35g/$\ell$-gel의 수준을 유지하였으며, 세포생존률도 약 70% 이상을 유지하였다.

• KSBB Journal
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• 제13권3호
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• pp.238-243
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• 1998

양어장 순환수 속의 암모니아성 질소의 제거를 위한 Ba-alginate 와 Ca-alginate 반응기는 0.6시간의 수력학적 체류시간에서 51.0, 52.6 g $NH_3-N/m^3/day$의 거의 유사한 암모니아성 집소의 제거속도릎 나타내었다. Ca-alginate를 이용한 합성양어장수 속의 암모니아성 짖소의 제거 실험에서 본 반응기의 암모니아성 짐소의 제거속도는 수력학적 체류시간이 0.3시간인 지점에서 82.0 g $NH_3-N/m^3/day$로 서 최고의 재거속도릎 나타냈으며 이때 암모니아 제거율은 48% 이었다. 수중의 암모니아의 농도가 2mg/L 정도의 범위에서는 반응기 내에 주입되는 공가량을 0.1 vvm 으로 공급하더라도 용존산소 농도를 7.0 - 5.6 g/m3로 유지 할 수 있었으므로 질화에 필요한 용존산소량을 충분히 유지할 수가 있는 것으로 나타났으며 pH 는 7.2 - 7.9의 범위플 유지할 수가 있어서 pH 변화에 따른 위 해 요소는 없는 것으로 나타났다.

• 한국식품과학회지
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• 제22권7호
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• pp.812-816
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• 1990

쏠비톨 생산을 위해서 Zymomonas mobilis의 Cell wall 투과성을 높인 후 alginate에 고정화 하였다. 투과성을 높이는데 toluene이 가장 효과적인 것으로 나타났으나, 3회의 회분공정을 행한 바 glucose-fructose oxidoreductase의 유출, 손실로 인하며 쏠비톨 생산이 급격히 저하되었다. 효소의 활성도를 장시간 유지하기 위하여 투과성을 높인 cell을 상온에서 1시간 동안 0.25%(v/v)의 glutaraldehyde로 처리한 후 alginate에 고정화 하여 연속공정을 행하였을 때 210시간 동안 높은 전환효율(82%)을 보였으며, 희석비율 $0.18\;h^{-1}$에서 $7.2{\sim}7.5\;g/l-h$의 쏠비톨 productivity를 얻었다. 이는 glutaraldehyde로 처리하지 않고 alginate에 cell을 고정화하였을 때에 비해서 효소의 안정성이 아주 높은 것으로 나타났다.

• Journal of Microbiology and Biotechnology
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• 제7권5호
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• pp.341-344
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• 1997

Batch cultures of Anacystis nidulans BD-1 resistant to azaleucine and fluorotyrosine produced and liberated a wide range of amino acids, notably glutamic acid, alanine, phenylalanine, leucine, isoleucine, cysteine and methionine. Sustained liberation for prolonged periods was achieved after immobilization on calcium alginate and the net concentration in the medium was 0.18-0.2 g $I^{-1}$. While acetohydroxy acid synthase in azaleucine-resistant mutant lost leucine- and isoleucine-sensitivity, fluorotyrosine-resistant strain turned phenylalanine activating. The activities of nitrate assimilating enzymes were also higher in the mutants and were relaxed from ammonium-repression. The metabolic adjustments involved in amino acid overproduction are discussed.

• Journal of Microbiology and Biotechnology
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• 제1권1호
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• pp.54-62
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• 1991

Cyclodextrin glucanotransferase(CGTase) derived from Bacillus macerans was immobilized by (1) covalent linkage on chitosan and chitin with glutaraldehyde, (2) adsorption on DEAE-cellulose and Amberite IRA 900 after succinylation, and (3) entrapment on alginate and polyacrylamide by cross linking. Adsorption on Amberite IRA 900 and covalent linking on chitosan were identified to be the most suitable immobilization methods considering the yield of activity and stability of immobilized CGTase. The enzymatic properties of immobilized CGTase were investigated and compared with those of the soluble CGTase. Thermal stability of CGTase immobilized on chitosan was increased from 50 to $55^{\circ}C$, and the optimum temperature of CGTase immobilized on Amberite IRA 900 was shifted from 55 to $50^{\circ}C$. The effect of molecular size of soluble starch (substrate) on immobilized CGTase investigated using partially liquefied substrates with different dextrose equivalent(DE). Cyclodextrin(CD) conversion yield augmented according to the increase of DE level for immobilized CGTase on Amberite IRA 900. CD conversion yield of partially cyclized starch with soluble CGTase was higher compared with liquefied one with ${\alpha}-amylase$.

• 약학회지
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• 제33권6호
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• pp.365-371
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• 1989

Microbiological conversion of sterols to 17-ketosteroids has been recognized as a source for commercial preparation of steroidal drugs. In order to develop bacterial strains and process with Brevibacterium lipolyticum IAM 1398 capable of converting cholesterol to 1,4-Androstadiene-3,17-dione (ADD) at about 27% yield, we studied on strain improvement, fermentation condition and whole cell immobilization. By using UV and/or NTG as mutagens, a mutant to convert cholesterol to ADD with higher yield than 60% was selected. Better production of ADD was manifested in the case of maltose used as a supplemental carbon source, and yeast extract or soytone as a nitrogen source. Addition of tween 80 (0.05%) as a surfactant beneficial for increasing the productivity. The optimal initial pH of the medium was 6.5 and optimal culture temperature was $30^{\circ}C$. Whole cell immobilization by using carrageenan, agar, alginate and acrylamide was carried out and the activity of conversion was tested. In the case of carrageenan and agar, immobilized cells were active for at least two cycles of fermentation.