• Title/Summary/Keyword: bio degradation

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Characterization of the Starch Degradation Activity of recombinant glucoamylase from Extremophile Deinococcus geothermalis (극한성 미생물Deinococcus geothermalis 유래 재조합 글루코아밀레이즈의 전분 분해 활성 특징)

  • Jang, Seung-Won;Kwon, Deok-Ho;Park, Jae-Bum;Jung, Jong-Hyun;Ha, Suk-Jin
    • Journal of Industrial Technology
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    • v.39 no.1
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    • pp.15-19
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    • 2019
  • This work focused on characterization of the starch degradation activity from extremophile strain Deinococcus geothermalis. Glucoamylase gene from D. geothermalis was cloned and overexpressed by pET-21a vector using E. coli BL21 (DE3). In order to characterize starch degrading activity of recombinant glucoamylase, enzyme was purified using HisPur Ni-NTA column. The recombinant glucoamylase from D. geothermalis exhibited the optimum temperature as $45^{\circ}C$ for starch degradation activity. And highly acido-stable starch degrading activity was shown at pH 2. For further optimization of starch degrading activity with metal ion, various metal ions ($AgCl_2$, $HgCl_2$, $MnSO_4{\cdot}4H_2O$, $CoCl_2{\cdot}6H_2O$, $MgSO_4$, $ZnSO_4{\cdot}7H_2O$, $K_2SO_4$, $FeCl_2{\cdot}4H_2O$, NaCl, or $CuSO_4$) were added for enzyme reaction. As results, it was found that $FeCl_2{\cdot}4H_2O$ or $MnSO_4{\cdot}4H_2O$ addition resulted in 17% and 9% improved starch degrading activity, respectively. The recombinant glucoamylase from D. geothermalis might be used for simultaneous saccharification and fermentation (SSF) process at high acidic conditions.

Inhibition of Polyphosphate Degradation in Synechocystis sp. PCC6803 through Inactivation of the phoU Gene

  • Han-bin Ryu;Mi-Jin Kang;Kyung-Min Choi;Il-Kyu Yang;Seong-Joo Hong;Choul-Gyun Lee
    • Journal of Microbiology and Biotechnology
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    • v.34 no.2
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    • pp.407-414
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    • 2024
  • Phosphorus is an essential but non-renewable nutrient resource critical for agriculture. Luxury phosphorus uptake allows microalgae to synthesize polyphosphate and accumulate phosphorus, but, depending on the strain of algae, polyphosphate may be degraded within 4 hours of accumulation. We studied the recovery of phosphorus from wastewater through luxury uptake by an engineered strain of Synechocystis sp. with inhibited polyphosphate degradation and the effect of this engineered Synechocystis biomass on lettuce growth. First, a strain (∆phoU) lacking the phoU gene, which encodes a negative regulator of environmental phosphate concentrations, was generated to inhibit polyphosphate degradation in cells. Polyphosphate concentrations in the phoU knock-out strain were maintained for 24 h and then decreased slowly. In contrast, polyphosphate concentrations in the wild-type strain increased up to 4 h and then decreased rapidly. In addition, polyphosphate concentration in the phoU knockout strain cultured in semi-permeable membrane bioreactors with artificial wastewater medium was 2.5 times higher than that in the wild type and decreased to only 16% after 48 h. The biomass of lettuce treated with the phoU knockout strain (0.157 mg P/m2) was 38% higher than that of the lettuce treated with the control group. These results indicate that treating lettuce with this microalgal biomass can be beneficial to crop growth. These results suggest that the use of polyphosphate-accumulating microalgae as biofertilizers may alleviate the effects of a diminishing phosphorous supply. These findings can be used as a basis for additional genetic engineering to increase intracellular polyphosphate levels.

Effect of Heat, Pressure, and Acid Treatments on DNA and Protein Stability in GM Soybean (GM 콩 DNA와 단백질의 안정성에 대한 열, 압력 및 산 처리의 영향)

  • Pack, In-Soon;Jeong, Soon-Chun;Yoon, Won-Kee;Park, Sang-Kyu;Youk, Eun-Soo;Kim, Hwan-Mook
    • Korean Journal of Food Science and Technology
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    • v.36 no.4
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    • pp.677-682
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    • 2004
  • Debates on safety of genetically modified (GM) crops have led to mandatory-labeling legislation of GM foods in many countries including Korea. Effects of heat, pressure, and acid treatments on degradation of DNAs or proteins in GM soybean at levels below detection limits of qualitative PCR and lateral flow strip test (LFST) methods were examined. Results showed that genomic DNAs and proteins were degraded into fragment sizes no longer possible for detection of inserted gene depending on thermal, or thermal and pressure treatment period. Detectaability of LFST for toasted meal increased in weakly treated soybean. DNA and protein detection methods were barely effective for detection of GM ingredient after $121^{\circ}C$ and 1.5 atmospheric treatment for 20 min. These results will be useful in determining GM labeling requirements of processed foods.

Treatment of Organic Wastes and Reuse of Bio-energy from the Anaerobic Digestion - Thermophilic Oxic Precess (혐기성 소화-고온 호기법에 의한 유기성폐기물의 처리와 생성열의 재활용 검토)

  • Yang, Jae-Kyung;Choi, Kyung-Min
    • Journal of the Korea Organic Resources Recycling Association
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    • v.9 no.1
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    • pp.79-89
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    • 2001
  • Anaerobic Digestion - Thermophilic Oxic Process(ADTOP) has been known to be one way reducing and composting of organic wastes without draining or forming excess sludge. It could be completely performed by the evaporation of water using the bio-energy from the microbial degradation of organic. In the present study the complete treatment of Chinese restaurant wastes was conducted and utility of bio-energy produced from the ADTOP was estimated. Base on results, it could be concluded as follows; 1) chinese restaurant wastes could be completely treated using the TOP without draining or excess sludge. Maximum volumetric loading rate was determined as $55.0kg-garbage/m^3$. Input water was almostly evaporated and 90.5% of carboneous organic wastes was conversed to carbondioxide. 2)The optimum volumetric loading rate which is acceptable to maintain over $55^{\circ}C$ in the anaerobic digester was determined as $45kg-garbage/m^3{\cdot}d$. 3) The optimum HRT was at least over 10 days in order to maintain about $50^{\circ}C$ in the anaerobic digester using bio-energy produced from TOP. Therefore the utilization of bio-energy produced from TOP could be used in the process which had long HRT such as the anaerobic digestion. 4) The efficiency of anaerobic digester rate were over 90% by the ADTOP under the organic loading rate of $1.1kg-COD/m^3{\cdot}d$, 50kg-Chinese restaurant garbage and $250{\ell}/m^3{\cdot}min$ of the aeration rate.

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Analysis of Expressed Sequence Tags from the Red Alga Griffithsia okiensis

  • Lee, Hyoung-Seok;Lee, Hong-Kum;An, Gyn-Heung;Lee, Yoo-Kyung
    • Journal of Microbiology
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    • v.45 no.6
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    • pp.541-546
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    • 2007
  • Red algae are distributed globally, and the group contains several commercially important species. Griffithsia okiensis is one of the most extensively studied red algal species. In this study, we conducted expressed sequence tag (ESTs) analysis and synonymous codon usage analysis using cultured G. okiensis samples. A total of 1,104 cDNA clones were sequenced using a cDNA library made from samples collected from Dolsan Island, on the southern coast of Korea. The clustering analysis of these sequences allowed for the identification of 1,048 unigene clusters consisting of 36 consensus and 1,012 singleton sequences. BLASTX searches generated 532 significant hits (E-value <$10^{-4}$) and via further Gene Ontology analysis, we constructed a functional classification of 434 unigenes. Our codon usage analysis showed that unigene clusters with more than three ESTs had higher GC contents (76.5%) at the third position of the codons than the singletons. Also, the majority of the optimal codons of G. okiensis and Chondrus crispus belonging to Bangiophycidae were G-ending, whereas those of Porphyra yezoensis belonging to Florideophycidae were G-ending. An orthologous gene search for the P. yezoensis EST database resulted in the identification of 39 unigenes commonly expressed in two rhodophytes, which have putative functions for structural proteins, protein degradation, signal transduction, stress response, and physiological processes. Although experiments have been conducted on a limited scale, this study provides a material basis for the development of microarrays useful for gene expression studies, as well as useful information for the comparative genomic analysis of red algae.

Application of a Modified Sublimation Method to Screen for PAH-Degrading Microorganisms (다환 방향족 탄화수소(PAH) 분해 미생물 탐색을 위한 승화법의 개발)

  • Kwon, Tae-Hyung;Kim, Jun-Tae;Kim, Jong-Shik
    • Korean Journal of Microbiology
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    • v.46 no.1
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    • pp.109-111
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    • 2010
  • Recent studies have described various microorganisms that can degrade PAH, however, there are currently limited methods available to screen for PAH-degrading microorganisms. To screen for PAH-degrading microorganisms, a sublimation method (Alley, Jeremy F. and Lewis R. Brown. 2000. Appl. Environ. Microbiol. 66, 439-442) was modified to produce a simple screening system. In our results, there were several bacterial species capable of pyrene degradation including genera, Coryenbacterium, Gordonia, Rhodococcus, and Streptomyces, which have been screened from 350 bacterial isolates of commercial gasoline and oil-spilled sediment by the sublimation method. The main advantage of this method is that it (i) safely deposits an even, thin and visible layer of PAH onto the agar surface without the use of solvents and (ii) the quantity of PAH sublimed onto the agar can be easily controlled. Overall, this sublimation method may be an effective and simple technique to screen for PAH-degrading microorganisms.

Validation and Recommendation of Methods to Measure Biogas Production Potential of Animal Manure

  • Pham, C.H.;Triolo, J.M.;Cu, T.T.T.;Pedersen, L.;Sommer, S.G.
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.6
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    • pp.864-873
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    • 2013
  • In developing countries, biogas energy production is seen as a technology that can provide clean energy in poor regions and reduce pollution caused by animal manure. Laboratories in these countries have little access to advanced gas measuring equipment, which may limit research aimed at improving local adapted biogas production. They may also be unable to produce valid estimates of an international standard that can be used for articles published in international peer-reviewed science journals. This study tested and validated methods for measuring total biogas and methane ($CH_4$) production using batch fermentation and for characterizing the biomass. The biochemical methane potential (BMP) ($CH_4$ NL $kg^{-1}$ VS) of pig manure, cow manure and cellulose determined with the Moller and VDI methods was not significantly different in this test (p>0.05). The biodegradability using a ratio of BMP and theoretical BMP (TBMP) was slightly higher using the Hansen method, but differences were not significant. Degradation rate assessed by methane formation rate showed wide variation within the batch method tested. The first-order kinetics constant k for the cumulative methane production curve was highest when two animal manures were fermented using the VDI 4630 method, indicating that this method was able to reach steady conditions in a shorter time, reducing fermentation duration. In precision tests, the repeatability of the relative standard deviation (RSDr) for all batch methods was very low (4.8 to 8.1%), while the reproducibility of the relative standard deviation (RSDR) varied widely, from 7.3 to 19.8%. In determination of biomethane concentration, the values obtained using the liquid replacement method (LRM) were comparable to those obtained using gas chromatography (GC). This indicates that the LRM method could be used to determine biomethane concentration in biogas in laboratories with limited access to GC.

Effects of Hormones on the Expression of Matrix Metalloproteinases and Their Inhibitors in Bovine Spermatozoa

  • Kim, Sang-Hwan;Song, Young-Seon;Hwang, Sue-Yun;Min, Kwan-Sik;Yoon, Jong-Taek
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.3
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    • pp.334-342
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    • 2013
  • Proteases and protease inhibitors play key roles in most physiological processes, including cell migration, cell signaling, and cell surface and tissue remodeling. Among these, the matrix metalloproteinase (MMPs) pathway is one of the most efficient biosynthetic pathways for controlling the activation of enzymes responsible for protein degradation. This also indicates the association of MMPs with the maturation of spermatozoa. In an attempt to investigate the effect of MMP activation and inhibitors in cultures with various hormones during sperm capacitation, we examined and monitored the localization and expression of MMPs (MMP-2 and MMP-9), tissue inhibitors of metalloproteinases (TIMP-2 and TIMP-3), as well as their expression profiles. Matured spermatozoa were collected from cultures with follicle-stimulating hormone (FSH), luteinizing hormone (LH), and Lutalyse at 1 h, 6 h, 18 h, and 24 h. ELISA detected the expression of MMP-2, MMP-9, TIMP-2, and TIMP-3 in all culture media, regardless of medium type (FSH-supplemented fertilization Brackett-Oliphant medium (FFBO), LH-supplemented FBO (LFBO), or Lutalyse-supplemented FBO (LuFBO)). TIMP-2 and TIMP-3 expression patterns decreased in LFBO and LuFBO. MMP-2 and MMP-9 activity in FBO and FFBO progressively increased from 1 h to 24 h but was not detected in LFBO and LuFBO. The localization and expression of TIMP-2 and TIMP-3 in sperm heads was also measured by immunofluorescence analysis. However, MMPs were not detected in the sperm heads. MMP and TIMP expression patterns differed according to the effect of various hormones. These findings suggest that MMPs have a role in sperm viability during capacitation. In conjunction with hormones, MMPs play a role in maintaining capacitation and fertilization by controlling extracellular matrix inhibitors of sperm.

Inhibitor Design for Human Heat Shock Protein 70 ATPase Domain by Pharmacophore-based in silico Screening

  • Lee, Jee-Young;Jung, Ki-Woong;Kim, Yang-Mee
    • Bulletin of the Korean Chemical Society
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    • v.29 no.9
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    • pp.1717-1722
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    • 2008
  • The 70 kDa heat-shock protein (Hsp70) involved in various cellular functions, such as protein folding, translocation and degradation, regulates apoptosis in cancer cells. Recently, it has been reported that the green tea flavonoid (−)-epigallocatechin 3-gallate (EGCG) induces apoptosis in numerous cancer cell lines and could inhibit the anti-apoptotic effect of human Hsp70 ATPase domain (hATPase). In the present study, docking model between EGCG and hATPase was determined using automated docking study. Epi-gallo moiety in EGCG participated in hydrogen bonds with side chain of K71 and T204, and has metal chelating interaction with hATPase. Hydroxyl group of catechin moiety also participated in metal chelating hydrogen bond. Gallate moiety had two hydrogen bondings with side chains of E268 and K271, and hydrophobic interaction with Y15. Based on this docking model, we determined two pharmacophore maps consisted of six or seven features, including three or four hydrogen bonding acceptors, two hydrogen bonding donors, and one lipophilic. We searched a flavonoid database including 23 naturally occurring flavonoids and 10 polyphenolic flavonoids with two maps, and myricetin and GC were hit by map I. Three hydroxyl groups of B-ring in myricetin and gallo moiety of GC formed important hydrogen bonds with hATPase. 7-OH of A-ring in myricetin and OH group of catechin moiety in GC are hydrogen bond donors similar to gallate moiety in EGCG. From these results, it can be proposed that myricetin and GC can be potent inhibitors of hATPase. This study will be helpful to understand the mechanism of inhibition of hATPase by EGCG and give insights to develop potent inhibitors of hATPase.

Bragg Reflecting Waveguide Device Fabricated on a Flexible Substrate using a Nano-imprinting Technology (나노임프린팅 기술을 이용한 유연성 브래그 반사 광도파로 소자)

  • Kim, Kyung-Jo;Yi, Jeong-Ah;Oh, Min-Cheol
    • Korean Journal of Optics and Photonics
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    • v.18 no.2
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    • pp.149-154
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    • 2007
  • Bragg reflecting waveguide devices have been fabricated on a flexible polymer substrate utilizing a post lift-off process which could Provide excellent uniformity of grating Patterns on Plastic film. The 510 m Period Bragg grating pattern is made by two methods. In the first sample the grating is fabricated by exposing the laser interference pattern on a photoresist, and then it is inscribed by $O_2$ plasma etching. The grating pattern of the second sample is formed by a PDMS soft mold imprinting process. The selective adhesion property of SU-8 material for Au and Si surfaces is utilized to prepare a 100-mm thick plastic substrate. Single mode waveguide is fabricated on the plastic substrate using polymer materials with refractive indices of 1.540 and 1.430 for the core and the cladding layers, respectively. The Bragg grating on Plastic substrate does not show any degradation in its spectral response compared to the reference sample made on a silicon wafer.