• Title/Summary/Keyword: Biological Engineering

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Quantitative Screening of Insect Cell Transformants Stably Expressing $GFP_{uv}-{\beta}1$, 3-N-acetylglucosaminyltransferase 2 Fusion Protein

  • Deo Vipin Kumar;Kato Tatsuya;Asari Naoko;Park Enoch Y.
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.3
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    • pp.275-279
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    • 2005
  • Insect cell transformants, stably expressing human $GFP_{uv}-{\beta}1$, 3-N-acetylglucosaminyltransferase 2 $({\beta}3GnT2)$ as the green fluorescent protein $(GFP_{uv})-fused$ protein, were efficiently isolated on Western blot by the quantification of the densitometric intensity of the fusion protein. From almost 150 transformants containing the fusion gene linked to three different types of signal sequence, two transformants, Tn-pXme4a and -pX28a, were successfully selected, showing 8.3 and 8.6 mU/mL ${\beta}3GnT$ activity, respectively. This method requires a screening time almost one-half that required in the isolation of stably transformed cells with high expression levels, and at the same time allows the handling a large number of transformants.

Enhancing Transfection Efficiency Using Polyethylene Glycol Grafted Polyethylenimine and Fusogenic Peptide

  • Lee, Haeshin;Jeong, Ji-Hoon;Lee, Je-Hoon;Park, Tae-Gwan
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.4
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    • pp.269-273
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    • 2001
  • This study presents a new formulation method for improving DNA transfection effi-ciency using a fusogenic peptide and polyethylene glycol-grafted polyethylenimine. Succinimidyls succinate polyethylene glycol (PEG-SSA) was conjugated with polyethylenimine(PEL). PEL is well known for a good endosomal escaping and DNA condensign agent. The positively charged syn-thetic fusogenic peptide, KALA was coated on the negatively charged PEG-g-PEI/DNA and PEI/DNA complexes. The KALA/PEI/ DNA complexes exhibited aggregation behavior at higher KALA coating amount with an effective diameter of around 1,000 nm. However, the LALA/PEG-g-PEI/DNA complexes were 100-300 nm in size with a surface zeta-potential (ζ)value of about +20mV. The conjugated PEG molecules suppressed any KALA-mediated inter-particle aggregation, and thereby improved the transfection efficiency, Consequently, the transfection efficiency of the KALA/PEG-g-PEI/DNA complexes was obtained by utilizing both the fusogenic activity of KALA and the steric repulsion effect of PEC.

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Structure-Based Virtual Screening of Protein Tyrosine Phosphatase Inhibitors: Significance, Challenges, and Solutions

  • Reddy, Rallabandi Harikrishna;Kim, Hackyoung;Cha, Seungbin;Lee, Bongsoo;Kim, Young Jun
    • Journal of Microbiology and Biotechnology
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    • v.27 no.5
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    • pp.878-895
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    • 2017
  • Phosphorylation, a critical mechanism in biological systems, is estimated to be indispensable for about 30% of key biological activities, such as cell cycle progression, migration, and division. It is synergistically balanced by kinases and phosphatases, and any deviation from this balance leads to disease conditions. Pathway or biological activity-based abnormalities in phosphorylation and the type of involved phosphatase influence the outcome, and cause diverse diseases ranging from diabetes, rheumatoid arthritis, and numerous cancers. Protein tyrosine phosphatases (PTPs) are of prime importance in the process of dephosphorylation and catalyze several biological functions. Abnormal PTP activities are reported to result in several human diseases. Consequently, there is an increased demand for potential PTP inhibitory small molecules. Several strategies in structure-based drug designing techniques for potential inhibitory small molecules of PTPs have been explored along with traditional drug designing methods in order to overcome the hurdles in PTP inhibitor discovery. In this review, we discuss druggable PTPs and structure-based virtual screening efforts for successful PTP inhibitor design.

Measurement of the Biological Active Point using the Bio-electrical impedance analysis based on the Adaptive Frequency Tracking Filter (적응주파수추적필터기반의 생체임피던스분석을 통한 생물학적활성점측정에 관한 연구)

  • Park, Hodong;Lee, Kyoungjoung;Yeom, Hojun
    • The Journal of the Institute of Internet, Broadcasting and Communication
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    • v.13 no.6
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    • pp.109-114
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    • 2013
  • The biological active points (BAP) are known as low resistance spots or good electro-permeable points. In this paper, a new method for BAP detection using the bio-impedance measurement system based on the adaptive frequency tracking filter (AFTF) and the transition event detector is presented. Also, the microcontroller process continuous time demodulation of the modulated signal by multi frequency components using the AFTF. The transition event detector based on the phase space method is applied about each frequency using the BAP equivalent model which is proposed.

Expression of Nutritionally Well-balanced Protein, AmA1, in Saccharomyces cerevisiae

  • Kim, Tae-Geum;Kim, Ju;Kim, Dae-Hyuk;Yang, Moon-Sik
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.3
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    • pp.173-178
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    • 2001
  • Food yeast, Saccharomyces cerevisiae, is a safe organism with a long history of use for the production of biomass rich in high quality proteins and vitamins. AmA1, a seed storage albumin from Amaranthus hypochondriacus, has a well-balanced amino acid composition and high levels of essential amino acids and offers the possibility of further improving food animal feed additives. In order to find an effective means of expressing AmA1 in yeast, the gene was cloned into an episomal shuttle vector. Four different promoters were tested: the glyceraldehyde-3-phosphate dehydrogenase promoter, galactose dehydrogenase 10 promoter, alcohol dehydrogenase II promoter, and a hybrid ADH2-GPD promoter. The recombinant AmA1 genes were then introduced into the yeast Saccharomyces cerevisiae 2805. Northern and Western blot analyses of the yeast under appropriate conditions revealed that AmA1 was expressed by all four promoters at varying levels. An enzyme-linked immunosorbent assay demonstrated that the amount of AmA1 protein in the recombinant yeast was 1.3-4.3% of the total soluble proteins. The highest expression level was obtained from the hybrid ADH2-GPD promoter.

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Nitritation Characteristics Depending on Influent Nitrogen Concentration in a Biological Aerated Filter (Biological Aerated Filter에서 유입 질소농도에 따른 아질산화 특성)

  • Yoo, Ik-Keun
    • Journal of Korean Society on Water Environment
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    • v.30 no.1
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    • pp.1-7
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    • 2014
  • The purpose of this study was to investigate the nitrification characteristics of biological aerated filter (BAF) packed with ceramic media, especially focusing on nitrite build-up during nitrification. When increasing the nitrogen load above $1.63kgNH_4{^+}-N/m^3{\cdot}d$, ammonium removal efficiency decreased to less than 60% and the nitrite ratio ($NO_2{^-}-N/NO_x-N$) of higher than 75% was achieved due to the inhibitory free ammonia (FA, $NH_3-N$) concentration and oxygen limitation. FA inhibition, however, is not recommended strategy to promote nitrite build-up since FA concentration in the reactor is coupled with decreased ammonium removal efficiency. Nitrite ratio in the effluent was also affected by aeration rate and influent ammonium concentration. Ammonium oxidation was enhanced at a higher aeration rate regardless of influent ammonium concentration but, the nitrite ratio was dependent on both aeration rate and influent ammonium concentration. While a higher nitrite ratio was obtained when BAFs were fed with $50mgNH_4{^+}-N/L$ of influent, the nitrite ratio significantly decreased for a greater influent concentration of $200-300mgNH_4{^+}-N/L$. Taken together, aeration rate, influent ammonium concentration and FA concentrations kept in the BAF were found to be critical variables for nitrite accumulation in the BAF system.

Application of Advanced Treatment Process for Nitrogen Compounds Removal of Industrial Waste-water (산업폐수 중의 질소 성분의 제거를 위한 고도처리 공정의 응용)

  • Bhang, Sung-Hun;Lim, Eun-Tae;Jeong, Gwi-Taek;Park, Jae-Hee;Park, Seok-Hwan;Kim, Seong-Jun;Park, Don-Hee
    • KSBB Journal
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    • v.23 no.6
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    • pp.541-545
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    • 2008
  • This paper was studied to research regarding the removal of contained nitrogen in industrial wastewater which uses the A2O4 advanced water treatment process. The field researches of two companies' wastewater occurred in each wastewater treatment site to apply the A2O4 process system, it was observed them for 20 days. As a result of the A2O4 system advanced wastewater process which applied an altitude control process obtained $10{\sim}76\;mg/L$, and 20 mg/L total nitrogen compound concentration in the two wastewater plants. In conclusion, it applied the A2O4 system in the two companies' wastewater system.

A Constitutive Inulase from a Bacillus (Bacillus 균이 생산하는 구성효소로서의 이눌라아제)

  • Uhm, Tai-Boong;Hong, Jai-Sik;Sohn, Hee-Suk;Park, Mun-Kuk;Byun, Si-Myung
    • Applied Biological Chemistry
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    • v.28 no.3
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    • pp.131-136
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    • 1985
  • Recently, we investigated characteristics of a bacterium which has never been reported as an inulin hydrolyzing microorganism. Several properties of the isolated microorganism were aerobic, rod typed, spore forming and Gram positive. According to the Bergey's manual, this bacterium tentatively appeared to be Becillus subtilis. This bacterium produced inulase constitutively in the media containing giucose, sucrose or cellulose without inulin as a sole carbon source. Also, inulase activities of the bacterium per unit culture volume showed 1.1 unit/ml comparable to 0.9 unit/ml of Kluyveromyces fragilis with relatively short culture time.

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Unrecorded species of Korean protozoans discovered through the project of 'Discovery of Korean Indigenous Species' II

  • Kyu-Seok, Chae;Jongwoo, Jung;Won Je, Lee;Gi-Sik, Min
    • Journal of Species Research
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    • v.11 no.4
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    • pp.335-342
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    • 2022
  • Through the 'Discovery of Korean Indigenous Species' project hosted by the National Institute of Biological Resources (NIBR), new or unrecorded species of various protozoans inhabiting the Korean Peninsula have been discovered. Samples were collected from marine, freshwater, and soil habitats in South Korea and all species were identified and reviewed by experts working on the respective taxonomic group. This study includes 17 unrecorded Korean protozoan species discovered through this project, which belong to four phyla: Amoebozoa Lühe, 1913, Cercozoa Cavalier-Smith, 1998, Euglenozoa Cavalier-Smith, 1981, and Ciliophora Doflein, 1901. Among them, three families (Rhogostomidae Dumack et al., 2017, Parauronematidae Small and Lynn, 1985, and Cyclidiidae Ehrenberg, 1838) and three genera (Rhogostoma Belar, 1921, Parauronema Thompson, 1967, and Cyclidium Müller, 1773) were reported for the first time in Korea. Unlike the previous paper of 2017, this study provides a comprehensive taxonomic account of each species (e.g., species name, collection site, synonyms, specimen vouchers, diagnoses, and figures). Additionally, all the species were assigned Korean names.

Functions and values of sulfated polysaccharides from seaweed

  • D. P. Nagahawatta;N. M. Liyanage;Thilina U. Jayawardena;Fengqi Yang;H. H. A. C. K. Jayawardena;M. J. M. S. Kurera;Fahe Wang;Xiaoting Fu;You-Jin Jeon
    • ALGAE
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    • v.38 no.4
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    • pp.217-240
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    • 2023
  • Sulfated polysaccharides (SPs) isolated from seaweed have emerged as remarkable bioactive compounds with a wide spectrum of biological activities and have substantial value in the scientific and industrial domains. The current study explores the diverse biological activities of SPs and their relationship with their structures. This aids in an in-depth examination of the multifaceted biological activities of SPs, including anticoagulant, anti-inflammatory, antiviral, antioxidant, and immunomodulatory properties, which underpin their potential health benefits. Furthermore, the current study explores the complicated properties of SPs, with their extraction methodologies and techniques for precise characterization. Elucidation of the commercial significance of SPs derived from brown, red, and green seaweed by highlighting their potential applications has emphasized their importance in human well-being. Further, this review emphasizes the challenges needed to overcome research and industrial innovations for SPs. Collaboration among researchers, industry stakeholders, and regulatory authorities can overcome these challenges and elevate the potential of SPs to revolutionize industries such as pharmaceuticals, cosmeceuticals, food, and biotechnology.