• 제목/요약/키워드: Microbial Biotechnology

Search Result 2,215, Processing Time 0.023 seconds

Isolation of lysozyme producing bacteria capable of solubilizing microbial cells (미생물 용해가 가능한 Iysozyme 분비 균주의 분리 및 특성)

  • Guo, Pengfei;Seo, Sun-Keun;Zhang, Lei;Kim, Hyo-Sang;Oh, Young-Khee;Jahng, Deok-Jin
    • KSBB Journal
    • /
    • v.23 no.3
    • /
    • pp.187-192
    • /
    • 2008
  • Lysozyme-producing microorganisms were isolated to obtain bacteria which can efficiently solubilize microbial cells. Cells of normal and chloroform-treated Escherichia coli and Micrococcus Iysodeikticus were used as model substrates to isolate lysozyme-producing microorganisms and investigate the efficiency of cell lysis. The culture supernatant of the isolate New1 (98% similarity of 16S rDNA sequence with Thermomonas haemolytica) showed different lytic characteristics for different substrates. Thermal treatment (autoclave) of substrate cells showed a significant effect on cell solubilization by culture supernatant of the New1. For autoclaved substrate cells, E. coli, M. Iysodeikticus and chloroform-treated E. coli were solubilized by 58.7%, 49.4% and 79.1%, respectively, in the culture supernatant of New1. The lytic activity of New1 was mainly caused by lysozyme produced by the isolate. It was also showed that New1 exhibited high protease activity and a little cellulase activity.

Transgenic Lettuce Expressing Chalcone Isomerase Gene of Chinese Cabbage Increased Levels of Flavonoids and Polyphenols

  • Han, Eun-Hyang;Lee, Ji-Sun;Lee, Jae-Woong;Chung, In-Sik;Lee, Youn-Hyung
    • Horticultural Science & Technology
    • /
    • v.29 no.5
    • /
    • pp.467-473
    • /
    • 2011
  • Flavonoid are large group of the polyphenolic compounds which are distinguished by an aromatic or phenolic ring structure and the phenolic compounds are induced by microbial infection, ultraviolet radiation, temperature and chemical stress. They are known for their antioxidant activity, anti-allergic, anti-inflammatory, anti-microbial and anti-cancer activities. In this study, changes in flavonoid content were investigated using heterologous chalcone isomerase (CHI) expression system. Also, phenolic compounds level was measured to examine the relation between flavonoids and phenols contents. Explants of lettuce (Lactuca sativa L.) were transformed with Agrobacterium tumefaciens LBA 4404 strain containing pFLH-CHI (derived from pPZP2Ha3) vector constructed with CHI gene from Brassica rapa. The putative transgenic plants were confirmed by genomic DNA PCR analysis. Also the transcription levels of the gene were analyzed by semi-quantitative RT-PCR with gene specific primers. The total flavonoid contents were increased at $T_0$ and $T_1$ generations over 1.4 and 4.0 fold, respectively. Total phenol contents also increased at $T_1$ generation. These results indicate that CHI gene plays an important role to regulate the accumulation of flavonoids and its component changes.

Induction of Lysozyme Gene Expression During Involution of Mouse Mammary Gland

  • Lee, M.J.;Han, O.;Back, K.;Choi, Y.J.;Baik, M.G.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.14 no.4
    • /
    • pp.462-466
    • /
    • 2001
  • To understand molecular mechanisms of mouse mammary gland involution, clones were isolated by differential screening of a cDNA library. Partial sequences of a clone showed 100% identity to cDNA sequences of mouse lysozyme P gene. Northern analysis was performed to examine expression levels of lysozyme mRNA in mammary gland at several physiological states. Expression of lysozyme gene was induced at involution day 5 compared with lactating stage. High levels of lysozyme mRNA were also detected at virgin tissues. Two types of separate genes, P and M lysozyme, have been known in mouse, and we found that both lysozyme P and M genes were expressed in mammary tissues by reverse transcriptase-polymerase chain reaction. The lysozyme enzyme activity determined by lysoplate assay was also higher in involuted mammary tissues compared with lactating tissues, showing a similar trend to its mRNA levels. Lysozyme is an antimicrobial protein and involved in host defense mechanism. The increase in lysozyme gene expression may help to prevent microbial infection during mammary gland involution at which stage the residual milk in the mammary gland provides good nutritional sources for microbial growth.

Degradation of Phenanthrene by Bacterial Strains Isolated from Soil in Oil Refinery Fields in Korea

  • KIM JEONG DONG;SHIM SU HYEUN;LEE CHOUL GYUN
    • Journal of Microbiology and Biotechnology
    • /
    • v.15 no.2
    • /
    • pp.337-345
    • /
    • 2005
  • The degradation of phenanthrene, a model PAH compound, by microorganisms either in the mixed culture or individual strain, isolated from oil-contaminated soil in oil refmery vicinity sites, was examined. The effects of pH, temperature, initial concentration of phenanthrene, and the addition of carbon sources on biodegradation potential were also investigated. Results showed that soil samples collected from four oil refinery sites in Korea had different degrees of PAH contamination and different indigenous phenanthrene-degrading microorganisms. The optimal conditions for phenanthrene biodegradation were determined to be 30$^{circ}C$ and pH 7.0. A significantly positive relationship was observed between the microbial growth and the rate of phenanthrene degradation. However, the phenanthrene biodegradation capability of the mixed culture was not related to the degree of PAH contamination in soil. In low phenanthrene concentration, the growth and biodegradation rates of the mixed cultures did not increase over those of the individual strain, especially IC10. High concentration of phenanthrene inhibited the growth of microbial strains and biodegradation of phenanthrene, but was less inhibitory on the mixed culture. Finally, when non-ionic surfactants such as Brij 30 and Brij 35 were present at the level above critical micelle concentrations (CMCs), phenanthrene degradation was completely inhibited and delayed by the addition of Triton X100 and Triton N101.

Characterization of the Microbial Diversity in a Korean Solar Saltern by 16S rRNA Gene Analysis

  • Park, Soo-Je;Kang, Cheol-Hee;Rhee, Sung-Keun
    • Journal of Microbiology and Biotechnology
    • /
    • v.16 no.10
    • /
    • pp.1640-1645
    • /
    • 2006
  • We studied the diversity of the halophilic archaea and bacteria in crystallizer ponds of a Korean solar saltern by analyzing 16S rRNA gene libraries. Although diverse halophilic archaeal lineages were detected, the majority (56%) were affiliated with the uncultured and cultured Halorubrum group. Halophilic archaea that have been frequently observed in solar saltern environments previously, such as Halogeometricum, Halococcus, Haloarcula, and Haloferax, were not detected in our samples. The majority of clones (53%) belonged to the Cytophaga-Flavobacterium-Bacteroides and ${\alpha}-,\;{\gamma}-,\;and\;{\delta}-Proteobacteria$ groups, with 47% of the clones being affiliated with ${\gamma}-Proteobacteria$. We also identified new ${\delta}-Proteobacteria$-related bacteria that have not been observed in hypersaline environments previously. Our data show that the diversity of the halophilic archaea and bacteria in our Korean saltern differs from that of solar salterns found in other geographic locations. We also showed by quantitative real-time PCR analysis that bacteria can form a significant component of the microbial community in solar salterns.

Overexpression of YbeD in Escherichia coli Enhances Thermotolerance

  • Kim, Sinyeon;Kim, Youngshin;Yoon, Sung Ho
    • Journal of Microbiology and Biotechnology
    • /
    • v.29 no.3
    • /
    • pp.401-409
    • /
    • 2019
  • Heat-resistant microbial hosts are required for bioprocess development using high cell density cultivations at the industrial scale. We report that the thermotolerance of Escherichia coli can be enhanced by overexpressing ybeD, which was known to encode a hypothetical protein of unknown function. In the wild-type E. coli BL21(DE3), ybeD transcription level increased over five-fold when temperature was increased from $37^{\circ}C$ to either $42^{\circ}C$ or $46^{\circ}C$. To study the function of ybeD, a deletion strain and an overexpression strain were constructed. At $46^{\circ}C$, in comparison to the wild type, the ybeD-deletion reduced cell growth half-fold, and the ybeD-overexpression promoted cell growth over two-fold. The growth enhancement by ybeD-overexpression was much more pronounced at $46^{\circ}C$ than $37^{\circ}C$. The ybeD-overexpression was also effective in other E. coli strains of MG1655, W3110, DH10B, and BW25113. These findings reveal that ybeD gene plays an important role in enduring high-temperature stress, and that ybeD-overexpression can be a prospective strategy to develop thermotolerant microbial hosts.

Production of 4-Ethyl Malate through Position-Specific Hydrolysis of Photobacterium lipolyticum M37 Lipase

  • Lim, Chae Ryeong;Lee, Ha young;Uhm, Ki-Nam;Kim, Hyung Kwoun
    • Journal of Microbiology and Biotechnology
    • /
    • v.32 no.5
    • /
    • pp.672-679
    • /
    • 2022
  • Microbial lipases are used widely in the synthesis of various compounds due to their substrate specificity and position specificity. 4-Ethyl malate (4-EM) made from diethyl malate (DEM) is an important starting material used to make argon fluoride (ArF) photoresist. We tested several microbial lipases and found that Photobacterium lipolyticum M37 lipase position-specifically hydrolyzed DEM to produce 4-EM. We purified the reaction product through silica gel chromatography and confirmed that it was 4-EM through nuclear magnetic resonance analysis. To mass-produce 4-EM, DEM hydrolysis reaction was performed using an enzyme reactor system that could automatically control the temperature and pH. Effects of temperature and pH on the reaction process were investigated. As a result, 50℃ and pH 4.0 were confirmed as optimal reaction conditions, meaning that M37 was specifically an acid lipase. When the substrate concentration was increased to 6% corresponding to 0.32 M, the reaction yield reached almost 100%. When the substrate concentration was further increased to 12%, the reaction yield was 81%. This enzyme reactor system and position-specific M37 lipase can be used to mass-produce 4-EM, which is required to synthesize ArF photoresist.