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Rhodobacter sphaeroides에 의한 ${\delta}-aminolevulinic{\;}acid$생산에 있어서 glutamic acid 및 감마 유도체의 영향
최경민,임왕진,황세영,Choi, Kyung-Min,Lim, Wang-Jin,Hwang, Se-Young 한국응용생명화학회 1993 Applied Biological Chemistry (Appl Biol Chem) Vol.36 No.3
The effect of ${\delta}-aminolevulinic\;acid$ (ALA) biosynthetic precursors and related compounds on the ALA productivity from a strain of Rhodobacter sphaeroides has been examined in vivo and in vitro systems. The relative ratios of ALA productivities by $C_{4}$- pathway to that by $C_{5}$-pathway in vivo and in vitro systems were 0.78 and 1.37, respectively. Although the expression rates of $C_{4}-$ and $C_{5}-pathways$ in cell-free systems prepared after precursors supplemented cultivations were increased 1.35 and 1.52 folds, respectively, the rate increase of $C_{4}-pathway$ was accompanied by the rate decrease of the $C_{5}-pathways$, and vice versa, as that the rates of both $C_{4}-$ and $C_{5}-pathways$ were lowered to be 0.91, 0.83, respectively. The order of cellular uptake rates of ${\gamma}-glutamyl$ derivatives relative to that found with L-glutamic acid were shown to be D-glutamic acid, 0.55: D-glutamine, 0.5: L-glutamine, 0.4: ${\gamma}-L-glutamyl$ ethylester, 0.3: GSH and Glu-pNA, 0. L and D configurations of glutamine were indicated as better substrates in vivo for ALA yields than those of glutamic acid, respectively. Rhodobacter sphaeroides 균주가 생산하는 ${\delta}-aminolevulinic\;acid(ALA)$의 생산성에 관하여 in vivo, in vitro 상에서 기질 및 관련 화합물을 이용하여 검토하였다. $C_5\;ALA$ 생합성계에 의한 ALA yield 대비의 $C_4$ 생합성계에 의한 비율은 in vive 상에서 0.78인 반면, in vitro 상에서의 비율은 1.37이었다.$C_4\;C_5$ 각 계의 기질 첨가 배양에 의한 cell-free system의 $C_4,\;C_5$ 계의 발현도는 미첨가 배양에 의한 system과 비교하여 각각 1.35, 1.52로 증가하였으나, 증가한 계에 대한 상대계의 발현도가 억제되어, $C_4$ 와 $C_5$ 계가 각각 0.91, 0.83으로 나타났다. ${\gamma}-Glutamyl\;derivatives$의 세포내 uptake rates는 L-glutamic acid를 기준으로 비교해서 D-glutamic acid, 0.55: D-glutamine, 0.5: L-glutamine, 0.4: ${\gamma}-L-glutamyl\;ethylester$, 0.3: GSH 및 Glu-pNA, 0의 순서를 보였다. Uptake rate와 관계없이 in vivo 상에서 L-과 D-glutamine이 L-, D-glutamic acid보다 균체 외 ALA의 생산에 있어서 각각 높은 yield의 효과를 보였다.
Rhodocyclus gelatinosus KUP-74의 휴지균체를 이용한 ${\delta}-Aminolevulinate$의 생산
이소희,안영철,임왕진,황세영,Lee, So-Hee,An, Young-Chul,Lim, Wang-Jin,Hwang, Se-Young 한국응용생명화학회 1994 Applied Biological Chemistry (Appl Biol Chem) Vol.37 No.6
광합성세균인 Rhodocyclus gelatinosus KUP-74의 휴지균체에 의한 균체외 ${\delta}-aminolevulinate$(ALA) 생산 효율성에 관하여 검토하였다. 균체외 ALA의 최대 생산치를 위하여 1 ml ALA 생산계는 1.05 mg의 휴지균체로 6시간의 배양을 필요로 하였다. 또한, 이 조건에서 ALA 생산의 유도물질인 levulinate나 L-glutamate 등은 균체외 ALA 생산에 있어서 비교적 낮은 효과를 나타내었다. 반면에, ALA 생산계의 부피와 이에 적합한 균체밀도가 효과적인 ALA 생산을 위하여 중요한 요인으로 나타났다. 휴지균체의 안정한 ALA 생산유지를 위한 수명은 휴지균체를 고정화 시키므로써 크게 연장되었다. The effectiveness of the resting cells of a photosynthetic bacterium, Rhodocyclus gelatinosus KUP-74, was investigated on the production of extracellular ${\delta}-aminolevulinate$ (ALA). The ALA generating system with 1.05 mg cells per milliliter required 6 hr-incubation to obtain maximal yield of extracellular ALA. Under this condition ALA inducers, i.e., levulinate or L-glutamate showed relatively low effects to increase extracellular ALA production. Instead, the volume of the system and proper cell density appeared to be important factors for effective production of the ALA. The life-span of the resting cells was remarkably extended to maintain stable production of the ALA by immobilization of cells.
Rhodocyclus gelatinosus KUP-74의 고정화균체로부터 ${\delta}-Aminolevulinate$의 연속생산
이소희,김현호,윤순규,임왕진,황세영,Lee, So-Hee,Kim, Hyun-Ho,Yun, Sun-Kyu,Lim, Wang-Jin,Hwang, Se-Young 한국응용생명화학회 1995 Applied Biological Chemistry (Appl Biol Chem) Vol.38 No.1
Rhodocyclus gelatinosus KUP-74의 고농도 휴지균체로부터 ${\delta}-aminolevulinate$(ALA)를 연속적으로 생산하기 위한 최적조건을 검토하였다. 균체외 ALA양은 20 mg cells/ml의 균체농도까지는 농도 증가에 따른 rectangular hyperbola의 증가양식을 보였으나, 그 이상의 균체농도는 ALA 생산에 전혀 증가효과를 보이지 않았다. 20 mg cells/ml의 반응계에서, 균체외 ALA의 효과적 생산을 위한 첨가물질의 최저농도는 levulinate 4 mM과 L-glutamate 5 mM로 나타났으며, 배양 3시간후 ALA 양의 최대치를 보였다. 한편, Ca-alginate를 이용한 포괄법으로 고정화시킨 균체로부터 효과적으로 ALA를 생산하기 위한 첨가물질의 최적농도는 levulinate 6 mM과 L-glutamate 10 mM로 나타났으며, 배양 6시간째에 균체외 ALA 양의 최대치를 보였다. 또한, 이 조건 하에서 고정화균체와 고농도의 휴지균체에 의한 ALA의 연속생산성을 검토한 결과, 이들 균체에 의한 50% ALA 생산성은 각각 185시간과 100시간에서 나타났으며, 생산된 균체외 ALA의 회수를 위해서는 균체를 고정화시키는 방법이 효과적이었다. An optimal condition for the continuous production of ${\delta}-aminolevulinate$(ALA) was investigated using high concentrated resting cells of Rhodocyclus gelatinosus KUP-74. The increase of the amount of extracellular ALA versus the concentration of resting cells showed rectangular hyperbolic pattern until 20 mg cells/ml, but no further increase in the ALA amount by increasing its concentration was occurred. The highest yield of the extracellular ALA was observed after 3 hr of incubation of 1 ml reaction system containing 20 mg cells, 4 mM levulinate and 5 mM L-glutamate. On the other hand, the immobilized cells prepared by Ca-alginate inclusive method needed to incubate for 6 hr with 6 mM levulinate and 10 mM L-glutamate to give maximal yield of the extracellular ALA. In addition, under these conditions the resulted continuous productivities of the ALA by immobilized cells and highly concentrated resting cells were appeared 50 percent decreases after incubations for 185 hr and 100 hr, respectively, and the method of the cells to be immobilized was more efficient to recover the extracellular ALA produced.
유기폐수처리를 위한 Rhodospirillum rubrum P17 의 종균생산
조경덕(Kyung Dug Cho),강성옥(Seong Og Kang),임왕진(Wang Jin Lim),조홍연(Hong Yon Cho),양한철(Han Chul Yang) 한국응용생명화학회 1993 Applied Biological Chemistry (Appl Biol Chem) Vol.36 No.6
A photosynthetic bacterium strain P17 having high growth rate and assimilating ability of organic acids was isolated from several soil samples, which was identified as Rhodospirillum rubrum. Cultural conditions of the strain P17 were examined for the production of starter culture used in the treatment of organic waste water. The addition of organic acids mixture as carbon source containing 0.2% Na-acetate, 0.1% Na-propionate and 0.2%r Na-lactate and 0.1% of yeast extract as growth factor stimulated the cell growth. The maximal cell production was obtained at 30℃, pH 7.0, 2,500 lux of illumination and 50∼100 rpm of agitation. Under the optimal conditions of batch and fed-batch culture systems in a Jar fermentor, 5.17 g/ℓ and 7.93 g/ℓ of cells were obtained after 5 days of cultivation, respectively. In continuous culture system, the cell productivity was 0.206 g/ℓ/h at a dilution rate of 0.21 h^(-1). When R. rubrum P17 was cultivated in a soybean curd waste water, initial COD level(3,240 ㎎/ℓ) of the waste water was reduced to 250 ㎎/ℓ after 4 days of cultivation.
Rhodocyclus gelationous KUP - 74 의 휴지균체를 이용한 δ - Aminolevulinate 의 생산
이소희(So Hee Lee),안영철(Young Chul An),임왕진(Wang Jin Lim),황세영(Se Young Hwang) 한국응용생명화학회 1994 Applied Biological Chemistry (Appl Biol Chem) Vol.37 No.6
The effectiveness of the resting cells of a photosynthetic bacterium, Rhodocyclus gelatinosus KUP-74, was investigated on the production of extracellular δ-aminolevulinate (ALA). The ALA generating system with 1.05 ㎎ cells per milliliter required 6 hr-incubation to obtain maximal yield of extracellular ALA. Under this condition ALA inducers, i.e., levulinate or L-glutamate showed relatively low effects to increase extracellular ALA production. Instead, the volume of the system and proper cell density appeared to be important factors for effective production of the ALA. The life-span of the resting cells was remarkably extended to maintain stable production of the ALA by immobilization of cells.
Rhodocyclus gelatinosus KUP - 74 의 고정화균체로부터 δ - Aminolevulinate 의 연속생산
이소희(So He Lee),김현호(Hyun Ho Kim),윤순규(Sun Kyu Yun),임왕진(Wang Jin Lim),황세영(Se Young Hwang) 한국응용생명화학회 1995 Applied Biological Chemistry (Appl Biol Chem) Vol.38 No.1
An optimal condition for the continuous production of δ-aminolevulinate(ALA) was investigated using high concentrated resting cells of Rhodocyclus gelatinosus KUP-74. The increase of the amount of extracellular ALA versus the concentration of resting cells showed rectangular hyperbolic pattern until 20 ㎎ cells/㎖, but no further increase in the ALA amount by increasing its concentration was occurred. The highest yield of the extracellular ALA was observed after 3 hr of incubation of 1 ㎖ reaction system containing 20 ㎎ cells, 4 mM lewlinate and 5 mM L-glutamate. On the other hand, the immobilized cells prepared by Ca-alginate inclusive method needed to incubate for 6 hr with 6 mM levulinate and 10 mM L-glutamate to give maximal yield of the extracellular ALA. In addition, under these conditions the resulted continuous productivities of the ALA by immobilized cells and highly concentrated resting cells were appeared 50 percent decreases after incubations for 185 hr and 100 hr, respectively, and the method of the cells to be immobilized was more efficient to recover the extracellular ALA produced.
팽이버섯 균사체 생장에 미치는 Natural Oil의 영향
안장혁,임왕진,송치현,양한철 한국산업미생물학회 1993 한국미생물·생명공학회지 Vol.21 No.4
합성배지에 첨가된 모든 식물성유들이 균사생장을 촉진하여 2%(v/v)의 식물성유들에 의해 균체량이 최저 2.7배에서 최고 3.3배까지 증가하였으며, 3%의 미강유(ricebran oil)농도에서 건조균체량은 무첨가시보다 3.5배가 증가되었다. 또한 균사체의 lipid 이용은 Ca^(2+)에 의해 촉진되어 1.0%(w/v)의 CaCl_2 농도에서 균체량은 18.2 ㎎/㎖로 최대치를 나타내었다. Flask에서 17일 배양하여 얻어진 액체종균과 고체종균을 접종하여 생산된 자실체의 양은 각각 118.0 g, 115.0 g으로 큰 차이를 나타내지 않았다. Vegetable oils supplemented to the basal medium stimulated mycelial growth of Flammulina velutipes. The mycelial yield was increased 3.5 folds by addition f 3% (v/v) ricebran oil. Maximum mycelial yield (18.2㎎/㎖) was obtained by addition of 3.0% ricebran oil with 1.0% CaCl_2 to the basal medium. There was no significant difference between the liquid and solid spawn in the yield of sporophores.