• Title/Summary/Keyword: culture broth

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Anti-wrinkle Activity of $\beta$-carotene Extracted & Purified from Recombinant Escherichia coli (재조합 대장균으로부터 추출.정제된 베타-카로틴의 주름개선 활성)

  • Jo, Ji-Song;Ku, Bo-Mi;Kang, Sang-Soo;Lee, Jae-Ran;Kim, You-Geun;Lee, He;Kim, Sung-Bae;Kim, Seon-Won;Kim, Chang-Joon;Chung, In-Young
    • KSBB Journal
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    • v.23 no.6
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    • pp.513-518
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    • 2008
  • This paper described the extraction/purification of $\beta$-carotene from recombinant E.coli and evaluation of anti-wrinkle activity of purified $\beta$-carotene. No significant differences in extraction yields were observed when hexane or isobutyl acetate was used. However, extraction from wet-cell cake resulted in 2-fold higher amount of $\beta$-carotene than that from dry cells. Disruption of 5 g-wet cells by ultrasonic homogenizer, acetone dehydration, extraction with isobutyl acetate resulted in 36 mg of $\beta$-carotene corresponding to 61.2% of recovery. The formation and separation of $\beta$-carotene crystal improved the purity. 633 mg of $\beta$-carotene crystal with 93% purity was obtained from 223 g/L of wet-cell cake harvested from 2.5-L fed-batch culture broth. The cultures of normal human primary fibroblast were performed to investigate the effect of $\beta$-carotene on cytotoxicity as MTT assay and anti-wrinkle activity as collagen synthesis assays. $1.7{\mu}M$ of $\beta$-carotene was found to be optimal concentration at which 1.4-fold higher amount of collagen was synthesized than that in absence of $\beta$-carotene. This indicates that highly purified $\beta$-carotene can be obtained from recombinant E.coli by applying simple method with less toxic solvent and can be used in functional cosmetics as anti-wrinkle agent.

Characterization of a Novel Fibrinolytic Enzyme Produced from Bacillus subtilis BK-17 (Bacillus subtilis BK-17 유래 혈전용해 효소의 특성)

  • Hyun Bek;Lim Hak-Seob;Chung Kyung Kae;Choi Yung Hyun;Choi Byung Tae;Seo Min-Jeong;Kim Ji-Eun;Ryu Eun-Ju;Huh Man Kyu;Joo Woo Hong;Jeong Young Kee
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.987-993
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    • 2005
  • A bacterium, producing a fibrinolytic enzyme, was screened from a decaying rice plant. The bacterium was identified as Bacillus subtilis by morphological, biochemical, and physiological properties and named Bacillus subtilis BK-17. The fibrinolytic enzyme (BK) was purified from supernatant of Bacillus subtilis BK-17 culture broth. The molecular weight was 31 kDa as determined by SDS-PAGE. The effect of temperature, pH, and plasminogen on the activity of the bacillokinase (BK) was analysed and the activity was compared with urokinase. The optimal temperature and pH were $50^{circ}C$ and pH 7, pH 8, respectively. The BK activity was inhibited to $45\%$, $35\%$, and $23\%$ with 1mM EDTA, $Zn^{2+}$, and $Ca^{2+}$, respectively. However, $Mg^{2+}$, $Mn^{2+}$, and $Co^{2+}$ ions did not have any significant effect on the enzyme activity The BK showed the artivity in the both plates, plasminogen-free fibrin plate and plasminogen-rich fibrin plate. The result indicates that the BK can directly act the fibrin. In comparison of fibrinolytic activity with urokinase on the fibrin plate, the BK shows about 20 folds higher activity than that of the urokinase.

Effect of Ethanolic Extract of Schizandra chinensis for the Delayed Ripening Kimchi Preparation (오미자(Schizandra chinensis) 추출물이 김치의 과숙억제에 미치는 영향)

  • Moon, Young-Ja;Park, Sun;Sung, Chang-Keun
    • The Korean Journal of Food And Nutrition
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    • v.16 no.1
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    • pp.7-14
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    • 2003
  • This study mainly focused on to investigate the effects of Schizandra chinensis on the growth of a bacterium, CS6 which was isolated from kimchi. CS6 was final]y identified to lactobacillus plantarum that caused acidification of kimchi. The ethanolic extract of Schizandra chinensis(EES) inhibited the growth of L. plantarum. Minimum inhibition concentration of crude EES on L. plantarum was 62.5mg/$m\ell$. In broth culture, 5$\mu\textrm{g}$/$m\ell$ of EES completely inhibited the growth of L. plantarum during fermentation. The addition of 0.4% of EES has no apparent effect on quality including the taste and color on kimchi. It was expected that EES-containing kimchi could extend the period of preservation. Analysis of organic acids in water fractions of EES was carried out by HPLC. It is apparent that antimicrobial active fractions contained the highest concentration of succinic acid, a little tartaric acid and malic acid. Among these organic acids, succinic acid showed the strong inhibitory effect against L. plantarum CS6 in vitro. Succinic acid-containing kimchi with a concentration of 0.4 and 0.5% had the inhibitory effect on growth of L. plantarum. Inhibitory effect of EES on amylase, cellulase and pectinase was also tested. In conclusion, the present experiment demonstrated that EES inhibited the growth of L. plantarum, and various enzyme activity. EES-containing kimchi was sustained the hardness, and initial acidity during fermentation. EES was considered as the possible additive of kimchi process and EES added in kimchi increase the quality, and storage period of kimchi.

Isolation and Characterization of Lactobacillus Strains Isolated from Korean Feces (한국인에서 분리된 Lactobacillus 속 젖산균의 동정 및 생리적 특성)

  • Kang, Dong-Gyu;Kang, Seog-Pil;Chang, Dong-Hoon;Kim, Sang-Ho;Yoon, Sung-Sik
    • Korean Journal of Food Science and Technology
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    • v.33 no.5
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    • pp.567-573
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    • 2001
  • Lactobacilli have been generally recognized as an important lactic acid bacteria present in the normal human intestinal flora. Fifty two Lactobacillus isolates were recovered from the feces of healthy Koreans whose age ranged from thirteen days after birth to 37 years. Among the isolates above, 17 isolates were tentatively identified as strains of Lactobacillus acidophilus and 3 isolates as strains of Lactobacillus casei. For their characterization, these twenty isolates were subjected to the experiments for the resistance to the artificial gastric juice, pH2.5 and bile salt. Interestingly, 3 strains survived pH 2.5 after 3 hour incubation in the artificial gastric juice with more than 75% of survival rate. L. acidophilus a-4 had the highest survival rate of 100%. Four strains including L. acidophilus a-3 grew gradually in MRS broth in the presence of the artificial bile salt. Cholesterol assimilation was also tested for the 20 isolates. The result showed that cholesterol concentration of the medium was reduced by 10 Lactobacillus isolates with more than 60%, as compared to the control. L. acidophilus a-2 had the highest reduction rate of 77%. Judging from these data obtained in vitro, some isolates ware likely to reach the lower small intestine after consumption without a significant loss of viability, suggesting that they had the potential to be developed as a probiotic culture which might lower the cholesterol level in human.

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Characterization of Agarase from a Marine Bacterium Agarivorans sp. BK-1 (해양세균 Agarivorans sp. BK-1의 분리 및 β-아가라제의 특성 규명)

  • Ahn, Byeong-Ki;Min, Kyung-Cheol;Lee, Dong-Geun;Kim, Andre;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.29 no.11
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    • pp.1173-1178
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    • 2019
  • The purpose of this study was to isolate an agar-degrading marine bacterium and characterize its agarase. Bacterium BK-1, from Gwanganri Beach at Busan, Korea, was isolated on Marine 2216 agar medium and identified as Agarivorans sp. BK-1 by 16S rRNA gene sequencing. The extracellular agarase, characterized after dialysis of culture broth, showed maximum activity at pH 6.0 and $50^{\circ}C$ in 20 mM Tris-HCl buffer. Relative activities at 20, 30, 40, 50, 60, and $70^{\circ}C$ were 67, 93, 97, 100, 58, and 52%, respectively. Relative activities at pH 5, 6, 7, and 8 were 59, 100, 95, and 91%, respectively. More than 90% of the activity remained after a 2 hr exposure to 20, 30, or $40^{\circ}C$; about 60% of the activity remained after a 2 hr exposure to $50^{\circ}C$. Almost all activity was lost after exposure to 60 or $70^{\circ}C$ for 30 min. Zymography revealed three agarases with molecular weights of 110, 90, and 55 kDa. Agarose was degraded to neoagarobiose (46.8%), neoagarotetraose (39.7%), and neoagarohexaose (13.5%), confirming the agarase of Agarivorans sp. BK-1 as a ${\beta}$-agarase. The neoagarooligosaccharides generated by this agarase could be used for moisturizing, bacterial growth inhibition, skin whitening, food treatments, cosmetics, and delaying starch degradation.