• Title/Summary/Keyword: Microbial Biotechnology

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Structural Characteristics of Expression Module of Unidentified Genes from Metagenome (메타게놈 유래 미규명 유전자의 발현에 관련된 특성분석)

  • Park, Seung-Hye;Jeong, Young-Su;Kim, Won-Ho;Kim, Geun-Joong;Hur, Byung-Ki
    • KSBB Journal
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    • v.21 no.2
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    • pp.144-150
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    • 2006
  • The exploitation of metagenome, the access to the natural extant of enormous potential resources, is the way for elucidating the functions of organism in environmental communities, for genomic analyses of uncultured microorganism, and also for the recovery of entirely novel natural products from microbial communities. The major breakthrough in metagenomics is opened by the construction of libraries with total DNAs directly isolated from environmental samples and screening of these libraries by activity and sequence-based approaches. Screening with activity-based approach is presumed as a plausible route for finding new catabolic genes under designed conditions without any prior sequence information. The main limitation of these approaches, however, is the very low positive hits in a single round of screening because transcription, translation and appropriate folding are not always possible in E. coli, a typical surrogate host. Thus, to obtain information about these obstacles, we studied the genetic organization of individual URF's(unidentified open reading frame from metagenome sequenced and deposited in GenBank), especially on the expression factors such as codon usage, promoter region and ribosome binding site(rbs), based on DNA sequence analyses using bioinformatics tools. And then we also investigated the above-mentioned properties for 4100 ORFs(Open Reading Frames) of E. coli K-12 generally used as a host cell for the screening of noble genes from metagenome. Finally, we analyzed the differences between the properties of URFs of metagenome and ORFs of E. coli. Information derived from these comparative metagenomic analyses can provide some specific features or environmental blueprint available to screen a novel biocatalyst efficiently.

Pulsed Electric Fields: An Emerging Food Processing Technology-An Overview (PEF 처리에 의한 식품의 가공)

  • Jayaprakasha, H.M.;Yoon, Y.C.;Lee, S.K.
    • Journal of Animal Science and Technology
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    • v.46 no.5
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    • pp.871-878
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    • 2004
  • Pulsed electric fields(PEF) technology is one of the latest nonthermal methods of food processing for obtaining safe and minimally processed foods. This technology can be effectively explored for obtaining safe food with minimum effect on nutritional, flavor, rheological and sensory qualities of food products. The process involves the application of high voltage(typically 20 ${\sim}$ 80 kv/cm) to foods placed between two electrodes. The mode of inactivation of microorganism; by PEP processing has been postulated in term; of electric breakdown and electroporation. The extent of destruction of microorganisms in PEF processing depends mainly on the electric field strength of the pulses and treatment time. For each cell types, a specific critical electric field strength and specific critical treatment time are required depending on the cell characteristics and the type and strength of the medium where they have been present. The effect also depends on the types of microorganisms and their phase of growth. A careful combination of processing parameters has to be selected for effective processing. The potential applications of PEF technology are numerous ranging from biotechnology to food preservation. With respect to food processing, it has already been established that, the technology is non-thermal in nature, economical and energy efficient, besides providing minimally processed foods. This article gives a brief overview of this technology for food processing applications.

Expression of Anthrax Lethal Factor, a Major Virulence Factor of Anthrax, in Saccharomyces cerevisiae (Yeast내에서 탄저병 원인균인 Bacillus anthracis의 치사독소인 Lethal Factor 단백질 발현)

  • Hwang Hyehyun;Kim Joungmok;Choi Kyoung-Jae;Chung Hoeil;Han Sung-Hwan;Koo Bon-Sung;Yoon Moon-Young
    • Korean Journal of Microbiology
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    • v.41 no.4
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    • pp.275-280
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    • 2005
  • Anthrax is an infectious disease caused by the gram-positive bacterium, Bacillus anthracis. Anthrax toxin is a tripartite toxin comprising of protective antigen (PA), lethal factor (LF) and edema factor (EF). PA is the receptor-binding component, which facilitates the entry of LF or EF onto the cytosol. LF is a zinc-dependent metalloprotease, which is a critical virulence factor in cytotoxicity of infected animals. Therefore, it is of interest to develop its potent inhibitors for the neutralization of anthrax toxin. The first step to identify the inhibitors is the development of a rapid, sensitive, and simple assay method with a high-throughput ability. Much efforts have been concentrated on the preparation of powerful assays and on the screening of inhibitors using these system. In the present study, we have tried to construct anthrax lethal factor in yeast expression system to prepare cell-based high-throughput assay system. Here, we have shown the results covering the construction of a new vector system, subcloning of LF gene, and the expression of target gene. Our results are first trial to express LF gene in eukaryote and provide the basic steps in design of cell-based assay system.

Identification and Molecular Characterization of Three Isoforms of Iturin Produced by Endophytic Bacillus sp. CY22 (식물 내생균 Bacillus sp. CY22가 생성하는 iturin isoform의 분리 및 특성)

  • Cho, Soo-Jeong;Yun-Han-Dae
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.1005-1012
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    • 2005
  • Endophytic Bacillus sp. CY22 was previously isolated from the interior of balloon flower root and showed strong antifungal activity against phytopathogenic fungi such as Rhizoctonia solnni, Fusarium oxysporum, and Phythium ultimum. Many Bacillus strains produce antifungal compound such as iturin, fengycin, and mycosubtilin. We isolated and identified antifungal compound from cell supernatant of the endophytic strain. By the MALDI-TOF mass result, the antifungal compound was similar to the known antifungal lipopeptide iturin. It was found that the purified iturin had three isoforms with protonated masses of m/z 1,043.39, 1,057.42, and 1,071.42 and different structures in combination with $Na^{+}$ ion using MALDI-TOF MS. The ita22 gene, which transacylase gene is associated with production of antifungal iturin, had an open reading frame (ORF) of 1,200 bp encoding 400 amino acids. Results of deduced amino acids sequence homology search, Ita22 was homologous with FenF (BAB69697) of Bacillus subtilis 168.

Characteristics of Kiwifruit-Added Traditional Kochujang (키위 첨가 전통고추장의 품질 특성)

  • Kim, Young-Soo;Song, Geun-Seoup
    • Korean Journal of Food Science and Technology
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    • v.34 no.6
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    • pp.1091-1097
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    • 2002
  • In order to improve palatability and quality of traditional kochujang, kiwifruit (Actinidia deliciosa) was added to traditional kochujang fermented for 3 months. Physicochemical and microbial characteristics were investigated during fermentation at $30^{\circ}C$. Moisture contents of all treated kochujangs increased with increasing amount of kiwifruit added during fermentation. pH of kiwifruit-added kochujang was lower than that of the control kochujang, whereas no significant differences in titratible acidity was observed among all treatments. Crude protein and salt contents of kiwifruit-added kochujang were lower than those of the control, whereas no difference in crude fat contents were found. Ethanol and amino-nitrogen contents of 6 and 9% kiwifruit-added kochujang were higher than those of the control. Fructose contents of kiwi-added kochujangs were higher than that of the control. Major organic acids of kochujang were in the order of malic acid > citric acid > succinic acid > acetic acid > lactic acid > oxalic acid. Succinic acid level increased significantly during fermentation and was higher in kiwifruit-added kochujang than in the control. Bacterial cell counts of all treatments were not different, and viable cell count of yeast was slightly higher in kiwifruit-added kochujang only at the initial fermentation period. Sensory evaluation revealed that the addition of 9% kiwifruit was the optimum condition for improving kochujang quality.

Comparison of UV-Irradiation and High Hydrostatic Pressure Sterilization for Storage Stability of Carrot Juice (UV살균과 초고압살균에 의한 당근주스의 저장성 비교)

  • Oh, Nam Seok;Kim, Min Young;Jang, Gwi Young;Baek, So Yune;Joung, Mi Yeun;Kang, Tae Su;Lee, Junsoo;Jeong, Heon Sang
    • The Korean Journal of Food And Nutrition
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    • v.30 no.5
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    • pp.1113-1118
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    • 2017
  • The objective of this study was to investigate the non-thermal sterilization effect of methods such as high hydrostatic pressure (HHP) and UV-irradiation (specifically with regard to the storage stability and shelf-life of carrot juice. The microbial counts of the non-sterilized product increased from 5.51 to 7.34 log CFU/mL up to the fifth day, and then decreased to 5.46 log CFU/mL at six days. UV-irradiation was increased from 2.37 to 4.92 log CFU/mL. HHP was maintained under the 3 log CFU/mL. The pH of UV-irradiation and HHP was maintained withing the range of 6.29~6.30 and 6.20~6.22 during storage. However, the pH arising from non-sterilization decreased from 6.31 to 4.49. The color of non-sterilization changed significantly during storage, but UV-irradiation and HHP were similar during storage. The ${\beta}$-carotene content of non-sterilization was noted to have decreased from 269.45 to $65.19{\mu}g/mL$ during storage. The UV-irradiation and HHP decreased from 263.46 and $268.35{\sim}281.16{\mu}g/mL$ to 243.42 and $244.09{\sim}269.29{\mu}g/mL$, respectively. In conclusion, these findings suggest that HHP can be used for the pasteurization, or sterilization, of carrot juice and the optimal condition is two minutes.

Effects of dietary humic acid and enzymes on meat quality and fatty acid profiles of broiler chickens fed canola-based diets

  • Disetlhe, Amogelang R.P.;Marume, Upenyu;Mlambo, Victor;Hugo, Arno
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.5
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    • pp.711-720
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    • 2019
  • Objective: This study was conducted to assess the effect of potassium humate and enzymes (Xylanase+Amylase+Protease) inclusion in diets on carcass characteristics, meat quality and fatty acid profiles of broilers fed canola-based diets. Methods: Two hundred and twenty broilers randomly allotted to 5 dietary treatments: the control (commercial broiler diet); CM (17.5% canola meal inclusion); CMEnz (17.5% CM inclusion+0.3 g/kg Axtra XAP); CMPh (17.5% CM inclusion+1.5% Potassium Humate, PH); and CMEnzPh (17.5% CM inclusion+1.5% PH+0.3 g/kg Axtra XAP) were slaughtered at day 42 for assessment of carcass and meat quality parameters. Results: Diet had no effect on carcass traits apart from breast muscle weight and breast muscle index. The highest breast muscle weight was observed in broilers fed CMEnz ($487.6{\pm}17.5g$) followed by those fed the control diet ($474.37{\pm}17.5g$). Diet also had no significant dietary effect on pH, temperature, drip loss and shear force values of the breast muscle. However, diet significantly affected meat colour and water-holding capacity. Broilers in the control and CMPh groups ($52.94{\pm}0.67$ and $52.91{\pm}0.67$) had the highest (p<0.05) values for lightness ($L^*$), whilst those fed CMEnzPh had the lowest value ($47.94{\pm}0.67$). In contrast, CM group had the lowest (p<0.05) value for redness ($a^*$) with CMEnzPh group having the highest values. The proportion of polyunsaturated fatty acids (PUFAs), n-6 and n-3 fatty acids and the PUFA/saturated fatty acid ratio were increased in CM-based diets containing enzymes and humic acid. Conclusion: It can, therefore, be concluded that CM can be included in broiler diets in the presence of enzymes and humic acid with positive effects on meat quality and important fatty acids that are beneficial to the health of consumers.

NLRP3 Inflammasome in Neuroinflammatory Disorders (NLRP3 인플라마좀 작용 기전 및 신경 질환에서의 역할)

  • Kim, Ji-Hee;Kim, YoungHee
    • Journal of Life Science
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    • v.31 no.2
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    • pp.237-247
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    • 2021
  • Immune responses in the central nervous system (CNS) function as the host's defense system against pathogens and usually help with repair and regeneration. However, chronic and exaggerated neuroinflammation is detrimental and may create neuronal damage in many cases. The NOD-, LRR-, and pyrin domain―containing 3 (NLRP3) inflammasome, a kind of NOD-like receptor, is a cytosolic multiprotein complex that consists of sensors (NLRP3), adaptors (apoptosis-associated speck like protein containing a caspase recruitment domain, ASC) and effectors (caspase 1). It can detect a broad range of microbial pathogens along with foreign and host-derived danger signals, resulting in the assembly and activation of the NLRP3 inflammasome. Upon activation, NLRP3 inflammasome leads to caspase 1-dependent secretion of the pro-inflammatory cytokines IL-1β and IL-18, as well as to gasdermin D-mediated pyroptotic cell death. NLRP3 inflammasome is highly expressed in CNS-resident cell types, including microglia and astrocytes, and growing evidence suggests that NLRP3 inflammasome is a crucial player in the pathophysiology of several neuroinflammatory and psychiatric diseases, such as Alzheimer's disease, Parkinson's disease, multiple sclerosis, stroke, traumatic brain injury, amyotrophic lateral sclerosis, and major depressive disorder. Thus, this review describes the molecular mechanisms of NLRP3 inflammasome activation and its crucial roles in the pathogenesis of neurological disorders.

A Study on the Activities of Five Natural Plant Essential Oils on Atopic Dermatitis (자생식물 Essential Oil 5 종의 항 아토피피부염 활성 연구)

  • Jeong, Jeong-Hwa;Nguyen, Thao Kim Nu;Choi, Min-Jin;Nguyen, Ly Thi Huong;Shin, Heung-Mook;Lee, Byung-Wook;Yang, In-Jun
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.47 no.1
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    • pp.23-30
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    • 2021
  • This study is an experiment to evaluate the anti-atopy efficacy of five kinds of natural plant essential oils; Artemisia annua L. (AA), Citrus junos Sieb. ex TANAKA (CJ), Chrysanthemum boreale Makino (CB), Pinus koraiensis (PK), and Pinus densiflora for. erecta (PD). Through Agar diffusion test, five species of native plant essential oils were treated in a total of four strains, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Candida albicans. In order to invest the anti-inflammatory effect, five kinds of natural plant essential oils were treated in HaCaT cells-induced by TNF-α and IFN-γ (TI). AA, CJ, CB, PK and PD showed antibacterial effects on Candida albicans at a concentration of 10 mg/mL. We also found that the thymus and activation-regulated chemokine (TARC) expression was suppressed in 0.1 ㎍/mL of PK, 1 ㎍/mL of AA, CB, and PK. macrophage-derived chemokine (MDC) expression was suppressed in 1 ㎍/mL of AA and PK. IL-6 expression was suppressed in 0.1, 1 ㎍/mL of AA, PK in HaCaT cells. Hence it suggests that AA, CB, and PK have the anti-inflammatory effects, and it could contribute to atopic dermatitis relief by reducing the infiltration of immune cells to inflamed area.

Supplementation of Microalgae (Tetradesmus sp.) to Pre-Starter Diet for Broiler Chickens (초기 육계 사료 내 미세조류(Tetradesmus sp.) 첨가에 따른 장관환경 변화)

  • Rim, Chae Yun;Jung, Hui-su;An, Su Hyun;Joo, Sang Seok;Kim, Z-Hun;Kong, Changsu;Kim, Myunghoo
    • Korean Journal of Poultry Science
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    • v.49 no.2
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    • pp.125-137
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    • 2022
  • This study aimed to determine the effects of dietary microalgae (Tetradesmus sp. (TO)) on intestinal immunity and microbiota of pre-starter broilers. One hundred and twenty 1-day-old birds (Ross 308) were allocated to two dietary treatment groups with six blocks in a randomized complete block design. The two experimental diets consisted of a corn-soybean meal-based basal diet and a diet with 0.5% TO powder instead of cornstarch in the basal diet. After feeding the experimental diets for ten days, all birds' body weight and feed intake were measured, and representative eight birds were selected from each treatment group. Small intestinal lamina propria cells were isolated using flow cytometry to examine the frequency of immune cells. Cecal feces were harvested for 16s rRNA gut microbiota analysis and fecal IgA levels. Here, we found that 0.5% TO supplementation increased CD3+CD4+ T cells in the small intestine, but decreased CD3+CD8+ T cells in the small intestine. Gut microbial analysis showed that TO supplementation significantly increased the alpha diversity of the gut microbiome. Taxonomic analysis showed that TO treatment increased the abundance of Firmicutes and decreased that of Bacteroidetes at the phylum level. The distribution of Enterobacteriaceae containing many harmful bacteria at the family level, was lower in the TO group. In the LEfSe analysis, the TO group had a significantly enriched abundance of Agathobaculum at the genus level. Overall, results show that Tetradesmus sp. supplementation influences intestinal T-cell immunity and induces the expansion of beneficial gut microbes in pre-starter broiler chickens.