• Title/Summary/Keyword: extracellular production

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Influence of Extracellular Products from Haematococcus pluvialis on Growth and Bacteriocin Production by Three Species of Lactobacillus

  • Kim Jeong-Dong;Lee Choul-Gyun
    • Journal of Microbiology and Biotechnology
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    • v.16 no.6
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    • pp.849-854
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    • 2006
  • The effects of Haematococcus pluvialis extracellular products on microbial growth and bacteriocin production were investigated to improve bacteriocin synthesis during the growth cycle of Lactobacilli. Lactobacillus pentosus KJ-108, L. plantarum KJ-10311, and L. sakei KJ-2008 were cultured in MRS and enriched medium (ERM) with or without supplement of the extracellular products obtained from a late exponential phase culture of Haematococcus pluvialis in modified Bold's basal medium (MBBM). In both MRS and ERM, the extracellular products strongly enhanced the growth as well as the bacteriocin production of all the lactic acid bacteria tested. The enhancing effect was observed in ERM with pH adjusted at 5 and 6. In addition, some difference in growth effects with the extracellular products of H. pluvialis was observed between pH 5 and 6 in ERM, but no effect was observed in the minimal medium. The final biomass and the final concentration of bacteriocin activity were associated with the cell growth that was promoted by the extracellular products of H. pluvialis, and the enhanced cell growth of the three lactic acid bacterial strains induced the increase of the specific bacteriocin production. Therefore, bacteriocin production and activity were influenced by the addition of the extracellular products of H. pluvialis in the culture medium.

Production of Extracellular Water Insoluble ${\beta}-1,3-Glucan$ (Curdlan) from Bacillus sp. SNC07

  • Gummadi, Sathyanarayana N.;Kumar, Kislay
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.6
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    • pp.546-551
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    • 2005
  • [ ${\beta}-1,3-Glucan$ ] (curdlan) is a water-insoluble polysaccharide composed exclusively of ${\beta}-1,3\;linked$ glucose residues. Extracellular curdlan was mostly synthesized by Agrobacterium species and Alcaligenes faecalis under nitrogen-limiting conditions. In this study, we screened the microorganisms capable of producing extracellular curdlan from soil samples. For the first time, we reported Gram-positive bacterium Bacillus sp. SNC 107 capable of producing extracellular curdlan in appreciable amounts. The effect of different carbon sources on curdlan production was studied and found that the yield of curdlan was more when glucose was used as carbon source. It was also found that maximum production was achieved when the initial concentration of ammonium and phosphate in the medium was 0.5 and 1.9 g/L respectively. In this study the curdlan production was increased from 3 to 7g/L in shake flask cultures.

Effect of Extracellular Calcium on Vitellogenin Production in the Culture of Hepatocytes in the Rainbow Trout, Oncorhynchus mykiss

  • Yeo In-Kyu;Mugiya Yasuo;Chang Young Jin;Hur Sung Bum;Yoo Sung Kyu
    • Fisheries and Aquatic Sciences
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    • v.1 no.1
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    • pp.24-29
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    • 1998
  • Effect of extracellular calcium in vitellogenin (VTG) production in response to estradiol-17 $\beta$ $(E_2,\;2\times10^{-6}M)$ was examined in primary hepatocyte culture of rainbow trout, Onchorhynchus mykiss. Total calcium in estrogenized sera significantly increased, compared with the control, while diffusible calcium was insignificant. However, diffusible calcium in the incubation medium with $E_2$ was significantly reduced, compared with the control. The uptake of extracellular calcium by cultured hepatocytes signifIcantly increased 90 min after $E_2$ addition. Moreover, the accumulation of intracellular calcium increased in the cultures with $E_2$, regardless of the calcium concentrations in the incubation media. In addition, $E_2-primed $ VTG production was significantly decreased by withdrawal of E_2$ from the incubation medium. Moreover, VTG production by $E_2-primed$ hepatocytes was reduced by removing calcium from the incubation medium with or without $E_2$. These results suggest that the entry of extracellular calcium into the cytoplasm is an important step for VTG production in primary hepatocyte cultures in rainbow trout.

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The Influence of NaCl and Carbonylcyanide-m-Chlorophenylhydrazone on the Production of Extracellular Proteases in a Marine Vibrio Strain

  • Kim, Young-Jae
    • Journal of Microbiology
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    • v.42 no.2
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    • pp.156-159
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    • 2004
  • In general, the salinity of the ocean is close to 3.5% and marine vibrios possess the respiratory chain-linked Na$\^$+/ pump. The influence of sodium chloride and the proton conductor carbonylcyanide m-chlo-rophenylhydrazone (CCCP) on the production of extracellular proteases in a marine Vibrio strain was examined. At the concentration of 0.5 M, sodium chloride minimally inhibited the activity of extra-cellular proteases by approximately 16%, whereas at the same concentration, the producton of extra-cellular proteases was severely inhibited. On the other hand, the production of extracellular proteases was completely inhibited by the addition of 2 ${\mu}$M CCCP at pH 8.5, where the respiratory chain-linked Na$\^$+/ pump functions.

Influence of Temperature, Oxygen, m-Chlorophenylhydrazone Cerulenin, and Quinacrine on the Production of Extracellular Proteases in Bacillus cereus

  • Kim, Sam-Sun;Park, Yong-Ha;Rhee, In-Koo;Kim, Young-Jae
    • Journal of Microbiology and Biotechnology
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    • v.10 no.1
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    • pp.103-106
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    • 2000
  • Bacillus cereus KCTC 3674 excretes at least two kinds of extracellular proteases into the growth medium. Two major bands of the protease activity with molecular weights of approximately 100 and 38 kDa were obtained after gelatin-SDS-PAGE. The protease with a molecular weight of 38kDa was identified as an extracellular neutral (metallo-) protease. The neutral protease was quite thermostabile but labile to alkaline pH. On the contrary, the 100-kDa protease was thermolabile but stable to alkaline pH. The production of 38-kDa neutral protease was strongly affected by temperature, oxygen, carbonylcyanied m-chlorophenylhydrazone(CCCP) that was defined as a protonophofre, and cerulenin which inhibited lipid synthesis and caused changes in the membrane composition. On the other hand, the production of the 100-kDa protease was strongly affected by only temperature and cerulenin. Quinacrine (0.2 mM), which inhibits the penicillinase-releasing proteases of Bacillus licheniformis, had no effect, whatsoever, on the production of extracellular proteases in B.cereus KCTC 3674.

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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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    • v.18 no.6
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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).

Culture Conditions of Geobacillus kaustophilus DSM 7263 for Production of Thermophilic Extracellular Lipase (내열성 extracellular lipase 생산을 위한 Geobacillus kaustophilus DSM 7263의 배양조건)

  • Jeon, Sung-Jong;Kang, Hyun-Woo
    • Journal of Life Science
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    • v.20 no.6
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    • pp.902-906
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    • 2010
  • A producer of thermophilic extracellular lipase, Geobacillus kaustophilus DSM 7263, was selected from various microorganisms of the Geobacillus genus. We investigated optimum conditions for mass production of G. kaustophilus lipase. Among the different natural oil media, olive oil was optimal for enzyme production. The maximum amount of enzyme production was obtained when G. kaustophilus was grown in a medium containing 0.5% olive oil as a carbon source. The pH and temperature for optimal growth were pH 8.0 and $55^{\circ}C$, respectively, while the optimum pH and temperature for lipase production were pH 6.0 and $50^{\circ}C$, respectively. In the presence of $Mg^{2+}$ and $Mn^{2+}$, lipase production was dramatically enhanced by 247% and 157%, respectively, whereas enzyme production was inhibited by $Zn^{2+}$, $Cu^{2+}$, and $Cd^{2+}$. The addition of 0.1% (v/v) triton X-100 increased lipase production and cell growth when compared to the negative control.

Study of metabolite production conditions by using the resting cells of Rhodospirillum rubrum N-1 (Rhodospirillum rubrum N-1의 휴지균체를 이용한 균체 대사산물의 생산 조건 연구)

  • 최경민;양재경
    • Journal of environmental and Sanitary engineering
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    • v.14 no.3
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    • pp.107-115
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    • 1999
  • The effectiveness of resting cells of a photosynthetic bacterium, Rhodospirillum rubrum N-1, was investigated on the production of extracellular ${\delta}-aminolevulinic$ acid(ALA). The ALA generating system required 3hr-incubation in the presence of 10mg of resting cells per ml to obtain the maximal yield of extracellular ALA. and also, under this condition the effect of ALA inducers, i.e., 30mM levulinic acid (LA) and L-glutamic acid($C_5$ pathway precursor) was relatively higher than that of produced extracellular ALA($83{\mu}M$). The volume of system and proper cell density appeared to be important factors for the effective production of extracellular ALA.

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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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    • v.30 no.8
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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.

Optimal Production of Protease from Entomopathogenic Fungus Beauveria bassiana (곤충 병원성 곰팡이 Beauveria bassiana로부터 Protease의 최적 생산)

  • Ko, Hwi-Jin;Kim, Hyun-Kyu;Kang, Sun-Chul;Kwon, Suk-Tae
    • Applied Biological Chemistry
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    • v.39 no.6
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    • pp.449-454
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    • 1996
  • We investigated the optimal condition for the production of extracellular pretense(a cuticle-degrading pretense) from entomopathogenic fungus Beauveria bassiana(ATCC7159) in liquid medium by adding of gelatin, bovine serum albumin(BSA), casein and polypeptone. The optimal induction medium for production of extracellular pretenses is composed of 0.5% polypeptone, trace elements and 50 mM potassium phosphate(pH 6.0). In this condition, the production of extracellular pretenses increased rapidly after the 24hrs, peaking at the third day and there was little inductive effect in culture broth more than pH 7.0. The pretenses were inhibited by phenyl methyl sulfonyl fluoride(PMSF). High activity of pretense was showed both range of pH 8.5 and 11.5 and also detected by three different portions of slice gel derived from non-denaturing isoelectricfocusing gel. At least three different extracellular pretenses are produced in optimal production medium when polypeptone is used as the sole carbon and nitrogen source.

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