• Title/Summary/Keyword: Shake culture

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Marked effect of Cuscuta on puerarin accumulation in cell cultures of Pueraria tuberosa grown in shake flasks and a bioreactor

  • Goyal, Shaily;Sharma, Varsha;Ramawat, Kishan G.
    • Plant Biotechnology Reports
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    • v.5 no.2
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    • pp.121-126
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    • 2011
  • Isoflavonoid production in cell cultures of Pueraria tuberosa as influenced by an angiospermic parasite, Cuscuta reflexa, was studied. During the time course, maximum isoflavonoid content was recorded when Cuscuta elicitor was added on day 15 of culture. Among various concentrations of elicitor tried, $1g\;l^{-1}$ of Cuscuta elicitor was found to be the most effective. The optimized elicitation conditions were used in vessels of varying capacity where maximum yield of ${\sim}91mg\;l^{-1}$ of isoflavonoid was recorded in a 2-l bioreactor which was about 19% higher than the control cultures. In this case, puerarin content increased up to $11mg\;l^{-1}$ which was 580% higher that the value recorded in the control cultures. In the bioreactor, 8 days of elicitation was optimal for the high accumulation of isoflavonoid, giving productivity of ${\sim}4mg\;l^{-1}\;day^{-1}$. The study showed persistent high isoflavonoid yield even during scale-up. Use of a preparation of Cuscuta reflexa as an elicitor is reported for the first time. The increase in isoflavonoid content was elicitor dose-dependent and can be explored to trigger high yields of isoflavonoid/secondary metabolites in production.

Secretory Expression of Human Growth Hormone in Saccharomyces cerevisiae Using Three Different Leader Sequences

  • Hahm, Moon-Sun;Chung, Bong-Hyun
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.4
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    • pp.306-308
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    • 2001
  • A recombinant human growth hormone(hGH) was expressed as a secretory product in the yeast Saccharomyuces cerevisiae. There different leader sequences derived from the mating fac-tor $\alpha$1(MF$\alpha$1) inulinase and invertase were used to direct the secretion of hGH into the extracel-lular medium. Among three leader sequences tested, the inulinase leader sequence was found to be the most efficient in the secretory expression of hGH. In contrast, no hGH was detected in the ex-tracellular medium with the invertase leader sequence. After 48 h shake-flask culture, the yields of hGH secreted into th emedium by the invertase. MF$\alpha$1 inulinase and invertase leader sequences were approximately 0, 0.3 and 0.9 mg/L, respectively. The secretion efficiencies were also found to be 0, 3.8 and 13% for the invertase , MG$\alpha$1 and inulinase leader sequences, respectively.

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Optimization of Medium Composition and Cultivation Parameters for Fructosyltransferase Production by Penicillium aurantiogriseum AUMC 5605

  • Farid, Mohamed Abdel-Fattah Mohamed;Kamel, Zinat;Elsayed, Elsayed Ahmed;El-Deen, Azza Mohamed Noor
    • Journal of Applied Biological Chemistry
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    • v.58 no.3
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    • pp.209-218
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    • 2015
  • Fructooligosaccharides have been mainly produced by microbial fructosyltransferases (FTase) enzymes. The present work focuses on the optimization of medium composition and cultivation parameters affecting FTase produced by Penicillium aurantiogriseum AUMC 5605 in shake flask cultivation. FTase production was optimized in two steps using DeMeo's fractional factorial design. A 1.46-fold increase in FTase production (105.4 U/mL) was achieved using the optimized culture medium consisting of (g/L): sucrose, 600; yeast extract, 10; $K_2HPO_4$, 5; $MgSO_4{\cdot}7H_2O$, 0.5; $(NH_4)_2SO_4$, 1.0 and KCl, 0.5. The obtained results showed that the maximum FTase enzyme activity was produced at initial cultivation pH values ranging from 6.0-6.5, at agitation speed of 200 rpm and using vegetative fungal cells as inoculum. Moreover, results showed that optimization of medium composition and some cultivation parameters resulted in an increase of about 93.7% in the enzyme activity than the nonoptimized cultivation conditions after 96 h of cultivation. Additionally, maximum production and specific production rates recorded 2340 U/L/h and 102 U/L/h/g cells, respectively.

Isolation and Nitrogen Removal Characteristics of Heterotrophic Nitrification-Aerobic Denitrifying Bacteria, Stenotrophomonas sp. CW-4Y (종속영양 질산화- 호기적탈질 세균 Stenotrophomonas sp. CW-4Y의 분리와 질소제거 특성)

  • Lee, Eun Young;Lee, Chang Won
    • KSBB Journal
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    • v.29 no.1
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    • pp.72-80
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    • 2014
  • CW-4Y was identified as Stenotrophomonas sp. by morphological and physiological characteristics, and phylogenetic analysis of its 16S rDNA gene sequence. Nitrogen removal by CW-4Y was analyzed in relation to the ammonium concentration, presence of organic carbon, carbon source, and carbon-to-nitrogen ratio (C/N). Stenotrophomonas CW-4Y has heterotrophic nitrification and aerobic denitrification abilities. Stenotrophomonas CW-4Y utilized only glucose as carbon sources, and heterotrophic nitrification and aerobic denitrification were observed regardless of the type of nitrogen source. The maximum ammonium removal rate of CW-4Y was 80 $mg-N{\cdot}L^{-1}{\cdot}d^{-1}$ and its denitrification rate of 192 $mg-N{\cdot}L^{-1}{\cdot}d^{-1}$ at $NO_3{^-}-N$ (about 280 ppm) in shake culture experiments at a C/N ratio of about 15 was about 30 times higher than those of other bacteria with the same ability.

Evolutionary Operation (EVOP) to Optimize Whey-Independent Serratiopeptidase Production from Serratia marcescens NRRL B-23112

  • Pansuriya, Ruchir C.;Singhal, Rekha S.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.5
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    • pp.950-957
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    • 2010
  • Serratiopeptidase (SRP), a 50 kDa metalloprotease produced from Serratia marcescens species, is a drug with potent anti-inflammatory property. In this study, a powerful statistical design, evolutionary operation (EVOP), was applied to optimize the media composition for SRP production in shake-flask culture of Serratia marcescens NRRL B-23112. Initially, factors such as inoculum size, initial pH, carbon source, and organic nitrogen source were optimized using one factor at a time. The most significant medium components affecting the production of SRP were identified as maltose, soybean meal, and $K_2HPO_4$. The SRP so produced was not found to be dependent on whey protein, but rather was notably induced by most of the organic nitrogen sources used in the study and free from other concomitant protease contaminant, as revealed by protease inhibition study. In addition, experiments were performed using different sets of EVOP design with each factor varied at three levels. The experimental data were analyzed with a standard set of statistical formula. The EVOP-optimized medium, with maltose 4.5%, soybean meal 6.5%, $K_2HPO_4$ 0.8%, and NaCl 0.5% (w/v), gave a SRP production of 7,333 EU/ml, which was 17-fold higher than the unoptimized media. The application of EVOP resulted in significant enhancement of SRP production.

Production of Cellulase from Lignocellulosic Waste. (리그노셀룰로스계 폐기물을 이용한 Cellulase의 생산)

  • 강성우;이진석;김승욱
    • Microbiology and Biotechnology Letters
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    • v.30 no.1
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    • pp.98-102
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    • 2002
  • Lignocellulosic wastes available in abundance can be excellent substrates for the production of cellulase. Different types of substrates and various pretreatments were used to improve the production of cellulase. The steam-exploded wood chip gave the highest activities of FPase (0.84 IU/mL) and CMCase (6.5 IU/mL) in the shake-flask culture. In 30 L bioreactor the steam-exploded wood chip and residue after saccharification gave the FPase activity (0.72 IU/mL) and the CMCase activity (6.3 IU/mL), respectively, similar those obtained in lactose.

Production of Mannitol by Lactobacillus sp. KY-107 (Lactobacillus sp. KY-107에 의한 Mannitol의 생산)

  • 윤종원;강선철류병호송승구
    • KSBB Journal
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    • v.11 no.3
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    • pp.374-379
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    • 1996
  • The production of extracellular mannitot by an efficient mannitol-producing bacterium, Lactobacillus sp. KY-107 was studied in shake flask culture using the modified MRS medium. Maximum mannitol production was obtained with fructose as the sole carbon source. Within 95 hours of incubation, a final concentration of 70g/L of mannitol from 100g/L fructose was obtained with an indicated yield of 86% based on fructose consumed. However, higher concentrations of fructose could not effectively be transformed to mannitol due to a lack of osmotolerance. The strain produced no other polyols such as glycerol and sorbitol as by-products. Yeast extract was best nitrogen source and high levels of inorganic phosphate up to 10g/L did not show any detrimental effect for mannitol formation. Manganese ion played important role in both cell growth and mannitol production. The optimum culture temperature and initial pH were $35^{\circ}C$ and 6-8, respectively.

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Production of Fungal Lipid (Part IV) Effect of Cultural Conditions on the Growth and Lipid Accumulation of Mucor plumbeus (곰팡이 유지 생산에 관한 연구 (제 4 보) 배양조건이 Mucor Plumbeus의 유지 생산에 미치는 영향에 대하여)

  • 유진영;이형춘;신동화;서기봉
    • Microbiology and Biotechnology Letters
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    • v.10 no.2
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    • pp.87-93
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    • 1982
  • The cultural conditions of Mucor plumbeus FRI 0007 were investigated for the maximum production of felt and lipid. It was found that the lower the pH and the higher the incubation temperature, the higher accumulation of the felt and lipid. Shake culture rendered higher lipid accumulation and lower felt accumulation than static culture. Maximum production of felt and lipid content were 47.8 g/$\ell$ and 50.73%, respectively, when the organisms were static-cultured at a temperature of 37$^{\circ}C$ and pH of 3.5 for 25 days latroscan thinchrographic analysis showed that the higher amount of triglyceride was obtained when static-cultured at a low pH. Fatty acid composition of the microbial lipid was affected by the incubation temperature, types of nitrogen source and speed of agitation: lower degree of saturation was observed as the incubation temperature decreased and the speed of agitation increased. Fatty acids of monoglyceride and diglyceride were mainly palmitic and oleic acids and those of triglycerides were mainly palmitic, oleic acids.

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Relationship Between Morphology and Itaconic Acid Production by Aspergillus terreus

  • Gao, Qian;Liu, Jie;Liu, Liming
    • Journal of Microbiology and Biotechnology
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    • v.24 no.2
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    • pp.168-176
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    • 2014
  • The morphology of filamentous fungi closely correlates with the productivity in submerged culture. Using itaconic acid (IA) production by Aspergillus terreus as a research model, the quantitative relationship between the growth form of A. terreus and IA production was investigated. IA fermentation was scaled up from shake flasks to a 7 L stirred tank bioreactor based on the quantitative relationship. Our results demonstrated the following: (1) Three morphologies of A. terreus were formed by changing the inoculum level and shape of the flask. (2) Investigation of the effects of the three morphologies on broth rheology and IA production revealed the higher yield of IA on dry cell weight (DCW, IA/DCW) and yield of glucose on DCW (consumed glucose/DCW) were achieved during clump growth of A. terreus. (3) By varying the $KH_2PO_4$ concentration and culture temperature, the relationships between clump diameter and IA production were established, demonstrating that the yield of IA on DCW ($R^2$ = 0.9809) and yield of glucose on DCW ($R^2$ = 0.9421) were closely correlated with clump diameter. The optimum clump diameter range for higher IA production was 0.40-0.50 mm. (4) When the clump diameter was controlled at 0.45 mm by manipulating the mechanical stress in a 7 L fermentor, the yield of IA on DCW and yield of glucose on DCW were increased by 25.1% and 16.3%, respectively. The results presented in this study provide a potential approach for further enhancement of metabolite production by filamentous fungi.

Screening of ${\gamma}-Linolenic$ Acid-Producing Fungi (감마-리놀렌산 생산 곰팡이의 탐색)

  • Shin, Yong-Chul;Shin, Hyun-Kyung
    • Korean Journal of Food Science and Technology
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    • v.20 no.5
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    • pp.724-731
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    • 1988
  • For a study on the production of ${\gamma}-linolenic$ acid(GLA) by microorganisms, fifteen strains of Mucorales obtained from culture stocks and ten isolate strains were compared in their cell growth, lipid content, fatty acid composition and pellet size formed in shake flask culture. Among the fungi examined, the isolated fungus, designated as FB-354, was found to be the most suitable one for the production of GLA mainly due to its high contents of lipid, 29.9% of dry cell weight and GLA, 16.8% of the total fatty acids. The strain FB-354 was tentatively identified as Mucor sp. on the basis of morphological characteristics. Fungal oil produced by Mucor sp. FB-354 was fractionated into 81.1% of neutral lipid, 7.2% of glycolipid, and 11.8% of phospholipid. Although the GLA content in the phospholipid fraction was as high as 21.4%, most of the GLA was found in the neutral lipid fraction.

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