• Journal of Industrial and Engineering Chemistry
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• 제67권
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• pp.231-235
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• 2018

In this study, we demonstrate the conversion of starch to (R,R)-2,3-butanediol (2,3-BD) in a hybrid cell-free synthesis system containing a mixture of lysates derived from Escherichia coli (E. coli) and cyanobacteria. A sufficient pool of pyruvate required for the synthesis of 2,3-BD was generated by combining metabolic pathways of cyanobacteria and E. coli. Successful synthesis of 2,3-BD was achieved by additional modifications of the hybrid cell-free system with the enzymes required to convert pyruvate to 2,3-BD. The results demonstrate a new approach to harness biological pathways to expand the scope of cell-free metabolic engineering by cross-species combinations of cell lysates.

• Journal of Microbiology and Biotechnology
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• 제11권6호
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• pp.928-933
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• 2001

Approximately 0.1 mg/ml of polyvinyl alcohol (PVA) was degraded by the growing cell, Brevibacillus laterospours, for 30 h, and 0.2 mg/ml of PVA was degraded by the cell-free extract that was isolated from Brevibacillus laterosporus. Approximately $0.29{\mu}g$/ml of acetic acid was produced from PVA by using the cell-free extract as a catalyst for 40 min. $V_{max}\;and\;K_m$ value of purified PAV-degradation enzyme was 3.75g/l and 2.75 g/l/min in reaction with EDTA and 3.99 g/l and 2.98 g/l/min in reaction without EDTA, respectively. Molecular weight of the purified enzyme determined by SDS-PAGE was 63,000 Da. Alcohol dehydrogenase and aldehyde dehydrogenase activities were qualitatively detected on a native acrylamide gel by an active staining method, indicating the existence of the metabolic pathway to use PVA as a substrate.

• Journal of Species Research
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• 제9권4호
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• pp.448-454
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• 2020

Nine free-living heterotrophic flagellates were cultured from marine intertidal sediments and freshwater sediments from Korea. These species are described with uninterpreted records based on light microscopy of living cells and reported taxonomically for the first time from Korea. Diagnostics of these species are as follows; Notosolenus hemicircularis: 9-11.8 ㎛ long with flagellar reservoir, ventrally flattened and dorsally convex with hyaline semicircular collar around short anterior neck, and 8 ridges on cell surface. Thecamonas tranhens: 4.5-7.1 ㎛ long, plastic with proboscis comprising an anterior flagellum surrounded by membranous sleeve. Bodomorpha minima: 4.5-7.0 ㎛ long, rigid with small rostrum in anterior end and active anterior flagellum. Cercomonas hiberna: 5.6-10.9 ㎛ long, very plastic with pseudopodia, cytoplasmic strand and 1 or 2 contractile vacuoles. Cercomonas pellucida: 7.5-13 ㎛ long, plastic with pseudopodia, cytoplasmic strand and single contractile vacuole. With nucleus closely connected to basal bodies. Eocercomonas echina: 4.7-6.5 ㎛ long, plastic with pseudopodia, cytoplasmic strand and 1 or 2 contractile vacuoles. Paracercomonas astra: 5.7-7.3 ㎛ long, moderately metabolic with pseudopodia, cytoplasmic strand and 1 or 2 contractile vacuoles. Paracercomonas minima: 5-9 ㎛ long, metabolic with pseudopodia, cytoplasmic strand and single contractile vacuole. Paracercomonas producta: 6.1-9.9 ㎛ long, very metabolic with pseudopodia, long cytoplasmic strand and single contractile vacuole.

• Journal of Microbiology and Biotechnology
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• 제23권12호
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• pp.1747-1756
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• 2013

The aim of this paper was to study the effect of spent medium recycle on Spodoptera exigua Se301 cell line proliferation, metabolism, and baculovirus production when grown in batch suspension cultures in Ex-Cell 420 serum-free medium. The results showed that the recycle of 20% of spent medium from a culture in mid-exponential growth phase improved growth relative to a control culture grown in fresh medium. Although both glucose and glutamine were still present at the end of the growth phase, glutamate was always completely exhausted. The pattern of the specific glucose and lactate consumption and production rates, as well as the specific glutamine and glutamate consumption rates, suggests a metabolic shift at spent medium recycle values of over 60%, with a decrease in the efficiency of glucose utilization and an increase in glutamate consumption to fuel energy metabolism. Baculovirus infection provoked a change in the metabolic pattern of Se301 cells, although a beneficial effect of spent medium recycle was also observed. Both growth rate and maximum viable cell density decreased relative to uninfected cultures. The efficiency of glucose utilization was dramatically reduced in those cultures containing the lowest percentages of spent medium, whereas glutamine and glutamate consumption was modulated, thereby suggesting that infected cells were devoted to virus replication, retaining their ability to incorporate the nutrients required to support viral replication. Recycle of 20% of spent medium increased baculovirus production by around 90%, thus showing the link between cell growth and baculovirus production.

• Journal of Microbiology and Biotechnology
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• 제11권3호
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• pp.406-411
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• 2001

In a bacterial denitrification test with Pseudomonas sp. and anaerobic consortium, more nitrates and less substrate were consumed but less metabolic nitrite was produced under an anaerobic $H_2$ condition rather than under $N_2$ condition. In a bioelectrochemical denitrification test with the same organisms, the electrochemically reduced neutral red was confirmed to be a substitute electron donor and a reducing power like $H_2$. The biocatalytic activity of membrane-free bacterial extract, membrane fraction, and intact cell for bioelectrochemical denitrification was measured using cyclic voltammetry. When neutral red was used as an electron mediator, the electron transfer from electrode to electron acceptor (nitrate) via neutral red was not observed in the cyclic voltammogram with the membrane-free bacterial extract, but it was confirmed to gradually increase in proportion to the concentration of nitrate in that of the membrane fraction and the intact cell of Pseudomonas sp.

• Biomaterials and Biomechanics in Bioengineering
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• 제2권3호
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• pp.143-157
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• 2015

Successful integration of cementless femoral stems using porous surfaces relies on effective periimplant bone healing to secure the bone-implant interface. The initial stages of the healing process involve protein adsorption, fibrin clot formation and cell osteoconduction onto the implant surface. Modelling this process in vitro, the current work considered the effect of fibrin deposition on the responses of human mesenchymal stromal cells cultured on ferritic fibre networks intended for magneto-mechanical actuation of in-growing bone tissue. The underlying hypothesis for the study was that fibrin deposition would support early stromal cell attachment and physiological functions within the optimal regions for strain transmission to the cells in the fibre networks. Highly porous fibre networks composed of 444 ferritic stainless steel were selected due to their ability to support human osteoblasts and mesenchymal stromal cells without inducing untoward inflammatory responses in vitro. Cell attachment, proliferation, metabolic activity, differentiation and penetration into the ferritic fibre networks were examined for one week. For all fibrin-containing samples, cells were observed on and between the metal fibres, supported by the deposited fibrin, while cells on fibrin-free fibre networks (control surface) attached only onto fibre surfaces and junctions. Initial cell attachment, measured by analysis of deoxyribonucleic acid, increased significantly with increasing fibrinogen concentration within the physiological range. Despite higher cell numbers on fibrin-containing samples, similar metabolic activities to control surfaces were observed, which significantly increased for all samples over the duration of the study. It is concluded that fibrin deposition can support the early attachment of viable mesenchymal stromal cells within the inter-fibre spaces of fibre networks intended for magneto-mechanical strain transduction to in-growing cells.

• 한국미생물·생명공학회지
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• 제49권3호
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• pp.298-304
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• 2021

Photosynthetic conversion through cyanobacteria and microalgae is an increasingly serious concern in the global warming crisis. Many value-added substances are produced through strain improvement, and much research and development is being conducted to determine its potential as an actual industrial strain. Economic barriers throughout processing production can be overcome to produce value-added chemicals by microalgal strains. In this study, we engineered cyanobacteria strains for the photosynthetic production of squalene and confirmed the continuous cultivation of CO₂ and light conditions. The free-inducer system of gene expression was developed at the cyanobacterial strains. Then, the squalene production level and growth of the recombinant cyanobacteria were analyzed and discussed. For bio solar-cell factories, the ability to regulate genes based on the free-inducer gene expression system promotes metabolic engineering research and construction to produce value-added chemicals.

• 한국미생물·생명공학회지
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• 제46권3호
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• pp.225-233
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• 2018

In this exploratory study, eight types of microalgae from different phyla (Chlamydomonas reinhardtii, Chlorella species, Haematococcus pluvialis, Porphyridium purpureum, Porphyridium cruentum, Isochrysis species, Isochrysis galbana, and Pavlova lutheri) were tested for their antibacterial activities against eight target pathogenic bacterial strains. The agar well diffusion method and broth micro dilution assay were conducted to estimate the antibacterial activity. Microalgae cell-free supernatants, exopolysaccharides (EPS), water, and organic solvent extracts were used for inhibition analysis. EPS extracted from P. lutheri showed activity against Bacillus subtilis and Pseudomonas aeruginosa. Inhibition zone diameters of 14-20 mm were recorded on agar plates, while the minimum inhibitory concentrations in the broth micro dilution assay were $0.39-25mg\;ml^{-1}$. During this study, haptophyte microalgae, Isochrysis species, and P. lutheri extracts showed the highest activity against most of the tested pathogenic bacterial strains, while most of the extracts were active against the important foodborne pathogen P. aeruginosa. This study showed promising results regarding important microalgae phyla, which will further aid research related to extracts and exploitation of bioactive metabolic compounds in the food and pharmaceutical industries.

• 센서학회지
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• 제26권5호
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• pp.314-323
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• 2017

Indicators to quantitatively evaluate the body function may help to optimize the effectiveness of rehabilitation therapy for stroke patients. In this study, we analyzed the body composition in the paretic and nonparetic regions of stroke patients with hemiplegia caused by cerebral hemorrhage (7 cases) and cerebral infarction (13 cases) using multifrequency bioelectrical impedance. Specifically, we considered fat mass (FM), fat-free mass (FFM), FFMI index (FFMI), FM/FFM relation, body cell mass (BCM), basal metabolic rate (BMR), and BMR/FFM relation to evaluate the bodily function in the paretic and nonparetic regions. These values showed considerable differences according to grades determined by the stroke causes and the paralysis status. In the paretic regions, the FFM, FFMI, BCM, and BMR were low and the FM was high. In contrast, the nonparetic regions showed a high FFM and low FM. Furthermore, the paretic and nonparetic regions of all patients suitably fit a linear relation (slope: 22.17 kcal/day/kg) between BMR and FFM. Therefore, bio-electrical impedance measurements can be very useful to quantitatively assess paretic and nonparetic regions in hemiplegic stroke patients.

• Journal of Plant Biotechnology
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• 제41권4호
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• pp.169-179
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• 2014

세포내에는 비효소적 반응으로 활성산소류를 제거하는 소분자의 항산화 물질과 과산소와 하이드로페록사이드를 분해하는 효소들이 존재한다. 항산화 시스템은 자유라디칼과 활성산소류를 제거함으로써 산화스트레스로부터 세포 구성요소들을 보호하는 역할을 한다. 비효소적 항산화 물질은 지용성과 친수성이 있는데, 지용성 물질들은 세포막에 위치하며 과산화지질이 형성되는 반응을 억제한다. 카로테노이드와 비타민E, $CoQ_{10}$은 세포 내에서 주요 지용성 항산화 물질로써, 이들이 대량으로 축적된 주요 작물의 개발은 영양학적 가치가 높은 식품을 생산할 수 있다. 본 총설에서는 식물에서 카로테노이드와 비타민E, $CoQ_{10}$의 생합성 경로와 대사공학을 이용한 영양강화 연구 현황을 기술하였다.