• 제목/요약/키워드: culture broth

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하이브리도마의 고농도 배양과 포도당 농도가 MAb 생산성에 미치는 영향 (High Density Culture of KA112 Hybridoma and Effect of Glucose Concentration on MAb Productivity)

  • 박상재;최차용
    • KSBB Journal
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    • 제8권5호
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    • pp.478-482
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    • 1993
  • LSM을 이용하여 KA112 균주의 고농도 배양을 시도하였다. Separator로는 hollow fiber를 사용하였고 reactor로는 Celligen을 이용하였다. Wroking volume 1리터로 10일간 배양하여 최고 세포농도가 회분식 배양에 비하여 10배 이상 증가한 $2.1\times10^7$ cells/ml이었고, 항체의 농도는 4.5배 정도 높았다. 최고 feed rate에서 항체생산속도는 회분식 배양보 다 9배 높았으며 배양 중 glucose농도가 Ig/e 이상일 때 specific productivity가 증가하였고, 1 g/6 이하얼 때 세포성장은 영향을 받지 않으냐 spe­c cific prodictivity는 감소하였다.

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제빵에서 밀가루 Brew의 조성이 젖산균의 생육에 미치는 영향 (The Effect of Composition of Flour Brew on Growth and Activity of Lactic Acid Bacteria)

  • 조남지
    • 한국식품영양학회지
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    • 제11권6호
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    • pp.683-688
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    • 1998
  • This study was conducted to develop optimal composition of flour brew in order to economically utilize flour brew inoculated by lactic acid bacteria as a starter(mother sponge) in bread-making. Two flour brews were prepared ; one with flour and water, the other with flour, water and NaCl. Various nutrients were added to both flour brews and Lactobacilli deMan Rogosa and Sharpe (MRS) broth to investigate the effect of them on growth and activities of Streptococcus thermophilus, Lactobacillus brevis and their mixed culture in flour brews to be tested with incubation at 37$^{\circ}C$. The growth of Streptococcus thermophilus, Lactobacillus brevis and their mixed culture was stimulated by addition of NaCl with 0.85% concentratin and more by mixed culture than by single lactic acid bacteria, resulting in 3 hrs reduction in cultivation and more by mixed culture than by single lactic acid bacteria, resulting in 3 hrs reduction in cultivation time. the addition of 3% glucose to flour brew with NaCl was observed to enhance acid productioni by mixed culture. Yeast extract greatly affected growth and activities of mixed culture of lactic acid bacteria in flour brew with NaCl and its optimum level of this additive in flour brew with NaCl was approximately 1.0%. The optimal composition of flour brew for mixed culture of lactic acid bacteria was suggested as follows; flour 100g, water 300g, NaCl 3.46g, glucose 12.48g, yeast extract 3.46g.

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귤피 및 녹차추출물에서 배양한 소나무잔나비버섯(Fomitopsis pinicola) 균사체의 유리아미노산 및 다당류 함량 (Free Amino Acid and Polysaccharide Content of Submerged Mycelial Culture of Fomitopsis pinicola)

  • 장경호;신경옥;김순동
    • 한국식품저장유통학회지
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    • 제12권4호
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    • pp.379-386
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    • 2005
  • 천연배지에서 배양한 소나무잔나비버섯(Fomitopsis pinicola)균사체와 균체외 유리아미노산 및 다당류함량을 조사하였다. 배지로는 YM(yeast-malt broth), CP(citrus peel-broth) 및 GT 배지(green tea-broth)를 사용하였으며 $30^{\circ}C$, 150rpm에서 10일간 배양하였다. 균사체의 수율은 GT, CP 및 YM에서 각각 42.3, 34.2 및 $9.89\%$였으며, 균사체를 제거한 배양액의 탁도는 GT 0.14, CP 0.16, YM 0.22이었다. 균사체의 총 유리아미노산 함량은 YM 928.19 $mg\%$, CP 1060.53 $mg\%$, GT 764.83 $mg\%$였으며, 주 아미노산은 YM은 lysine, CP와 GT는 glutamic acid였다. 균체외 총 유리아미노산 함량은 YM 659.75 $mg\%$, CP 954.55 $mg\%$, GT 838.69 $mg\%$였으며, 주 아미노산은 YM, CP 및 GT 모두 glutamic acid였다. 균사체와 균체외 total 아미노산 유도체의 함량은 CP>GT>YM 순이었다. 균사체의 주 아미노산 유도체는 YM은 cystathionine, CP와 GT는 hydroxy proline이였다. YM, CP 및 GT의 균체외 주 아미노산 유도체는 hydroxy proline이였다. 균사체와 균체외 AIS 함량은 CP>GT>YM순이었다. 산가용성 다당류 함량은 GT($0.69\%$)>YM($0.39\%$)>CP($0.18\%$)순, 균체외 산가용성 다당류 함량은 GT($0.87\%$)>CP($0.69\%$)>YM($0.09\%$)순이었다. 균사체의 알칼리 가용성 다당류 함량은 CP($5.21\%$)>GT($5.18\%$)>YM($4.56\%$) 순, 균체외는 GT($6.79\%$)>YM($3.57\%$)>CP($3.01\%$)순으로, 균사체 함유 다당류는 산가용성($0.18{\sim}0.69\%$)에 비하여 알칼리가용성($4.56{\sim}5.21\%$)의 함량이 현저하게 높았다. CP 배지에서 배양한 균사체의 알칼리가용성 다당류를 겔 여과한 결과 hexose와 uronic acid로 구성된 분자량 500,000 dalton의 다당류와 분자량 10,000 dalton이하의 단백질이 결합한 단백 다당체로 추정되었다.

Effect of a Bacterial Grass Culture on the Plant Growth and Disease Control in Tomato

  • Lee, Yong Seong;Naing, Kyaw Wai;Kim, Kil Yong
    • 식물병연구
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    • 제23권4호
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    • pp.295-305
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    • 2017
  • This study aimed to investigate the plant growth-promoting and biocontrol potential of a grass culture with Paenibacillus ehimensis KWN8 on tomato. For this experiment, treatments of a chemical fertilizer (F), a bacterial grass culture (G), a 1/3 volume of G plus 2/3 F (GF), and F plus a synthetic fungicide (FSf) were applied to tomato leaves and roots. The result showed that the severity of Alternaria solani and Botrytis cinerea symptoms were significantly reduced after the application of the bacterial grass culture (G and GF) and FSf. In addition, root mortality in G and GF was lower compared to F. Tomato plants treated with G or GF had better vegetative growth and yield compared to F. Application of G affected the fungal and bacterial populations in the soil. In conclusion, treatment with a bacterial grass culture decreased disease severity and increased tomato growth parameters. However, there were no statistically significant correlations between disease occurrence and tomato yields. This experiment presents the possibility to manage diseases of tomato in an environmentally friendly manner and to also increase the yield of tomato by using a grass culture broth containing P. ehimensis KWN38.

맥각균(麥角菌)의 배양(培養)에 관한 연구(硏究)(IV) -한국산(韓國産) 맥각균(麥角菌)의 배양(培養)- (Studies on the Culture of Ergot Fungus(IV) -Culture of Korean Ergot Fungus-)

  • 김병각;심미자;최응칠;박영인
    • 생약학회지
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    • 제6권1호
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    • pp.9-13
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    • 1975
  • To isolate ergot strains which are capable of growing and producing alkaloids in submerged culture, strains were isolated from the sclerotia parasitizing the Graminae plants in Korea and the experiments of submerged culture of these strains yielded the following results: 1) The mycelia which were respectively isolated from the sclerotia parasitizing Agropyron semicostatum $N_{EES}$, Arundinella hirta $T_{ANAKA}$ var. ciliata $K_{OIDSUMI}$, Ischaemum anthephoroides $M_{IQ}$. var. eriostachyum $H_{ONDA}$, and Phleum pretense L. grew in Medium D by submerged culture. 2) When the strain of the ergot of Agropyron semicostatum was inoculated into six different nutrient solutions for submerged culture, its mycelium grew well in Media C, D and F, but produced alkaloids only in Medium C, indicating that Medium C is relatively suitable for the strain. 3) The extraction of the alkaloids from the culture broth by ether and T.L.C. analysis of the extract showed that it contained at least two types of alkaloids.

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혼합 젖산균을 이용한 밀가루 용액의 반복 유가식 발효 (Repeated Fed-Batch Fermentation of Wheat Flour Solution by Mixed Lactic Acid Bacteria)

  • 김상용;노봉수;오덕근
    • 한국식품과학회지
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    • 제29권2호
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    • pp.343-347
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    • 1997
  • Lactobacillus brevis, L. fermentum과 L. plantarum의 혼합젖산균을 사용하여 배양조건이 밀가루 용액의 발효에 미치는 영향을 살펴보았다. 온도변화에 따른 pH와 총적정산도의 변화를 살펴본 결과 pH의 감소와 총적정산도의 증가가 가장 크게 나타난 $35^{\circ}C$를 밀가루 발효의 최적온도로 선정하였다. 5L발효조에서 질소가스를 1.0 vvm으로 첨가한 혐기적 조건보다 공기를 첨가한 호기적 조건에서 총적정산도의 증가와 pH의 감소가 더 크게 나타났다. 최적 산소공급 조건을 찾기 위하여 통기량을 1.0 vvm으로 고정하고 교반속도를 달리하여 밀가루 용액의 발효한 결과 총적정산도가 가장 크게 나타난 교반속도 250 rpm에 해당되는 산소 전달속도상수 $60\;hr^{-1}$에서 최적이었다. 선정된 최적 배양조건에서 밀가루 발효액의 생산성을 높이기 위하여 반복비율을 변화시키면서 pH-stat를 이용한 반복 유가식 배양을 수행하였다. 반복비율이 증가할수록 반복 간격시간은 증가하였으나 밀가루 발효의 최대 작동시간은 감소하였다. 최적 반복비율을 결정하기 위하여 단위시간 당 배양부피 당 생산된 밀가루 발효액의 부피와 최대가동시간 동안 배양부피 당 생산된 총 밀가루 발효액의 부피를 살펴본 결과 20%의 반복비율에서 최대값을 나타내었다. 혼합젖산균을 이용한 밀가루용액의 유가식 배양에서 최적조건은 배양온도 $35^{\circ}C$, 통기량 1.0 vvm, 산소전달속도 $60\;hr^{-1}$, 반복비율 20%로 나타났다.

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곤충병원세균(Photorhabdus temperata ssp. temperata) 유래 곤충 면역 억제물질 생성 비교 연구를 통한 저렴한 세균 배지 선발 (Comparative Analysis of Immunosuppressive Metabolites Synthesized by an Entomopathogenic Bacterium, Photorhabdus temperata ssp. temperata, to Select Economic Bacterial Culture Media)

  • 서삼열;장호진;김건우;김용균
    • 한국응용곤충학회지
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    • 제49권4호
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    • pp.409-416
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    • 2010
  • 곤충병원세균인 Photorhabdus temperata ssp. temperata(Ptt)는 곤충의 면역반응을 억제시켜 피기생 곤충 체내에서 공생하는 기주 선충의 발육을 도모하게 된다. 또한 Ptt의 변역억제 활성은 Bacillus thuringiensis(Bt)의 병원성을 증가시킨다. 본 연구는 이러한 유용 곤충병원세균의 대량 생산을 위한 저렴한 배지를 선발하기 위해 수행되었으며, 두 연구용 배지(LB, TSB)와 저렴한 산업용 두 배지(MY, M2)를 상호 비교하였다 모든 배양액에 동일한 밀도의 Ptt를 접종하고 배양하였을 때 48 시간 이후 정지상이 나타났다. 그러나 연구용 배양액인 LB와 TSB에서 두 가지 산업용 배양액보다 정지상에서 높은 세균 밀도를 보였다. 네 가지 배지에서 증식된 Ptt 배양액은 모두 배추좀나방(Plutella xylostella) 3령충에 대한 Bt 병원성을 현격하게 제고시켰고, 이들 배지 종류에 따라 치아가 없었다. 네 가지 배양액에서 세균의 증식에 의해 생산되는 대시물질의 양과 배지별 생산되는 대사물질의 동일성을 확인하기 위해 헥산과 에틸아세테이트의 유기용매로 추출했다. 시간별 배양액의 유기용매 추출물질은 세균의 증식과 비슷하게 대사물질의 생산량에서도 증가하는 것을 알 수 있었다. 역상 HPLC를 이용하여 네 가지 세균 배양액 각각에서 대사물질을 분리하였고, 정량적으로 네 가지 대사물질이 서로 다른 배지에서 통계적으로 차이 없이 검출되었다. 본 연구는 비교적 저렴한 두 가지 산업용 배지가 유용 대사물질의 생성에 변화 없이 Ptt 세균을 저렴하게 배양할 수 있다고 제시하고 있다.

Extracellular 5-Aminolevulinic Acid Production by Escherichia coli Containing the Rhodopseudomonas palustris KUGB306 hemA Gene

  • Choi, Han-Pil;Lee, Young-Mi;Yun, Cheol-Won;Sung, Ha-Chin
    • Journal of Microbiology and Biotechnology
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    • 제18권6호
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    • pp.1136-1140
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    • 2008
  • The Rhodopseudomonas palustris KUGB306 hemA gene codes for 5-aminolevulinic acid (ALA) synthase. This enzyme catalyzes the condensation of glycine and succinyl-CoA to yield ALA in the presence of the cofactor pyridoxal 5'-phosphate. The R. palustris KUGB306 hemA gene in the pGEX-KG vector system was transformed into Escherichia coli BL21. The effects of physiological factors on the extracellular production of ALA by the recombinant E. coli were studied. Terrific Broth (TB) medium resulted in significantly higher cell growth and ALA production than did Luria-Bertani (LB) medium. ALA production was significantly enhanced by the addition of succinate together with glycine in the medium. Maximal ALA production (2.5 g/l) was observed upon the addition of D-glucose as an ALA dehydratase inhibitor in the late-log culture phase. Based on the results obtained from the shake-flask cultures, fermentation was carried out using the recombinant E. coli in TB medium, with the initial addition of 90 mM glycine and 120 mM succinate, and the addition of 45 mM D-glucose in the late-log phase. The extracellular production of ALA was also influenced by the pH of the culture broth. We maintained a pH of 6.5 in the fermenter throughout the culture process, achieving the maximal levels of extracellular ALA production (5.15 g/l, 39.3 mM).

Engineering of a Microbial Cell Factory for the Extracellular Production of Catalytically Active Phospholipase A2 of Streptomyces violaceoruber

  • Lee, Hyun-Jae;Cho, Ara;Hwang, Yeji;Park, Jin-Byung;Kim, Sun-Ki
    • Journal of Microbiology and Biotechnology
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    • 제30권8호
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    • pp.1244-1251
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    • 2020
  • Phospholipase A2 (PLA2) from Streptomyces violaceoruber is a lipolytic enzyme used in a wide range of industrial applications including production of lysolecithins and enzymatic degumming of edible oils. We have therefore investigated expression and secretion of PLA2 in two workhorse microbes, Pichia pastoris and Escherichia coli. The PLA2 was produced to an activity of 0.517 ± 0.012 U/ml in the culture broth of the recombinant P. pastoris. On the other hand, recombinant E. coli BL21 star (DE3), overexpressing the authentic PLA2 (P-PLA2), showed activity of 17.0 ± 1.3 U/ml in the intracellular fraction and 21.7 ± 0.7 U/ml in the culture broth. The extracellular PLA2 activity obtained with the recombinant E. coli system was 3.2-fold higher than the corresponding value reached in a previous study, which employed recombinant E. coli BL21 (DE3) overexpressing codon-optimized PLA2. Finally, we observed that the extracellular PLA2 from the recombinant E. coli P-PLA2 culture was able to hydrolyze 31.1 g/l of crude soybean lecithin, an industrial substrate, to a conversion yield of approximately 95%. The newly developed E. coli-based PLA2 expression system led to extracellular production of PLA2 to a productivity of 678 U/l·h, corresponding to 157-fold higher than that obtained with the P. pastoris-based system. This study will contribute to the extracellular production of a catalytically active PLA2.

Effect of Dissolved Oxygen Concentration and pH on the Mass Production of High Molecular Weight Pullulan by Aureobasidium pullulans

  • LEE, JI-HYUN;JEONG-HWA KIM;MI-RYUNG KIM;SUNG-MI LIM;SOO-WAN NAM;JIN-WOO LEE;SUNG-KOO KIM
    • Journal of Microbiology and Biotechnology
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    • 제12권1호
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    • pp.1-7
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    • 2002
  • The effects of DO and pH on the mass production of pullulan with high molecular weight and the morphology of A. pullulans ATCC 42023 were evaluated. A. pullulans showed a maximum production of pullulan (11.98 g/l) when the initial pH of the culture broth was 6.5 in a shake-flask culture. In a batch culture, the mixture of a yeast-like and mycelial cell forms was found at a pH of 4.5, and the maximum production of pullulan (13.31 g/l) was obtained. However, a high proportion of high molecular weight pullulan (M.W.>2,000,000) was produced at a pH of 6.5, with a yeast-like morphology. The maximum pullulan production yield ($51\%$) was obtained at a pH noncontrol (initial pH 6.5) and DO control (above $50\%$) condition. Pullulan degrading enzyme was activated when the pH of the broth was lower than 5.0 and the portion of low molecular weight pullulan was increased. The formation of a black pigment was observed at an initial stationary phase, at 40 h of fermentation. Therefore, the fermentation should be carried out in a pH noncontrol (initial pH of 6.5) and DO control (above $50\%$) condition, and should be harvested before reaching the stationary phase (around 40 h) for the production of high molecular weight pullulan.