• Journal of Microbiology and Biotechnology
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• 제16권6호
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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.

• Biotechnology and Bioprocess Engineering:BBE
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• 제10권6호
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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.

• Fisheries and Aquatic Sciences
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• 제1권1호
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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.

• Journal of Microbiology
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• 제42권2호
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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.

• Journal of Microbiology and Biotechnology
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• 제10권1호
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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.

• 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).

• 생명과학회지
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• 제20권6호
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• pp.902-906
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• 2010

고온성 균주로 알려진 Geobacillus 속의 다양한 균주로부터 내열성 extracellular lipase를 생산하는 G. kaustophilus DSM 7263를 선별하였다. 우리는 본 균주로부터 lipase를 대량생산하기 위한 최적 조건을 조사하였다. 배양 배지에 다양한 천연오일을 첨가한 결과, lipase의 최적 생장을 위한 탄소원으로는 0.5% 올리브 오일이 최적 조건으로 확인되었다. 본 균주의 생장을 위한 최적온도와 pH는 각각 $55^{\circ}C$와 8.0인 반면, lipase 생산을 위한 최적 온도와 pH는 각각 $50^{\circ}C$와 6.0을 나타내어 최적생육조건과는 다른 양상을 나타내었다. 금속이온에 대한 영향에 대해서는 배지에 $Mg^{2+}$과 $Mn^{2+}$을 첨가한 경우 각각 247%와 157%의 효소 생산이 증가한 반면, $Co^{2+}$, $Fe^{2+}$, $Ni^{2+}$, $Cu^{2+}$는 효소 생산을 저해 하였다. 또한 0.1% (v/v) triton X-100을 첨가하면 대조구에 비해 효소생산과 균의 생장이 모두 증가하는 것으로 나타났다.

• 환경위생공학
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• 제14권3호
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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.

• 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.

• Applied Biological Chemistry
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• 제39권6호
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• pp.449-454
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• 1996

곤충 병원성 곰팡이 Beauveria bassiana(ATCC7159)로부터 곤충 cuticle 분해 효소인 extracellular protease의 최적 생산 조건을 gelatin, vine serum albumin(BSA), casein 및 polypeptone 등을 첨가하여 조사하였다. 이 효소의 최적 생산 배양 조건은 미량원소를 함유한 0.5% Polypeptone, 50mM potassium phosphate pH 6.0이었다. 이 조건에서 pretense의 생산은 배양 1일 후부터 급격히 상승하여 배양 3일 이후 최대로 생산되었으며 배양액의 pH가 7.0 이상이 되면 효소 생산효과가 거의 없었다. 이 pretenses는 phenyl methyl sulfonyl fluoride (PMSF)에 저해받는다. 효소 활성은 pH 8.5와 11.5에서 높게 나타났다. 또한 비변성 isoeletricfocusing gel 전기영동 후 각 gel 절편의 활성을 측정한 결과 protease활성이 세부분에서 나타났다. 따라서 탄소원 및 질소원으로 polypeptone을 이용하여 생산되는 pretense는 세종류 이상인 것으로 판단된다.