• Journal of the Korean Society of Manufacturing Process Engineers
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• v.20 no.8
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• pp.33-41
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• 2021

Recently, many studies have been conducted on substituting fossil fuels with bio-refineries in existing industrial systems using biomass. Among the various bio-refineries, microalgae have received wide attention because it uses inorganic compounds to produce useful substances, which are extracted by a cell disruption process. Although numerous cell disruption methods exist, cell disruption efficiency has been studied by ultrasonic treatment. Ultrasound is a high-frequency (20 kHz or higher) sound wave and causes cell disruption by cavitation when passing through a solvent. In this study, we used the microalgal species Chlorella sp., which was cultured in a plate-type photobioreactor. The experiment was conducted using a continuous low-frequency processing device. The reduction of cells with time due to cell disruption was fitted using a logistic model, and optimum conditions for highly efficient cell disruption were determined by conducting experiments under multiple conditions.

• Journal of the Korean Society of Manufacturing Process Engineers
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• v.19 no.2
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• pp.111-118
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• 2020

Recently, bioenergy research using microalgae, one of the most promising biofuel sources, has attracted much attention. Cell disruption, which can be classified as physical or chemical, is essential to extract functional ingredients from microalgae. In this study, we investigated the cell disruption efficiency of Chlorella sp. using low-frequency non-focused ultrasound (LFNFU). This is a continuously physical method that is superior to chemical methods with respect to environmental friendliness and low processing cost. A flat panel photobioreactor was employed to cultivate Chlorella sp. and its growth curve was fitted both with Logistic and Gompertz models. The temporal change in cell reduction by cell disruption using LFNFU was fitted with a Logistic model. The experimental conditions that were investigated were the initial concentration of microalgal cells, relative amplitude of output ultrasound waves, processing volume of microalgal cells, and initial pH value. The optimal conditions for the most efficient cell disruption were determined through the various tests.

• Journal of the Korean Society of Manufacturing Technology Engineers
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• v.26 no.1
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• pp.1-5
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• 2017

This study investigated the mechanical tearing of a cell membrane using a nanostructured alumina filter for easy and quick mechanical cell disruption. Nanostructured alumina filters were prepared by a multi-step aluminum anodizing process and nanopore etching process. Six different types of nanostructures were formed on the surface of the nanoporous alumina filters to compare the mechanical cell disruption characteristics according to the shape of the nanostructure. The prepared alumina filter was assembled in a commercial filter holder, and then, NIH3T3 fibroblast cells in a buffer solution were passed through the nanostructured alumina filter at a constant pressure. By measuring the concentration of proteins and DNA, the characteristics of mechanical cell disruption of the nanostructured alumina filter were investigated.

• BMB Reports
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• v.35 no.4
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• pp.428-431
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• 2002

The often-encountered problem of disrupting bacteria for the purpose of extracting soluble protein has generated various methods. Many require specialized equipment. Very often, especially during preliminary studies, investigators need a simple, fast, and inexpensive method for cell disruption that preserves biological activity. This paper compares some simple and inexpensive methods for cell disruption, such as bead-vortexing, freesing-thawing, French pressing, and sonication. It also provides some tips to increase protein yield and preserve biological activity. If performed under optimal conditions, bead-vortexing gives protein yields that are comparable to French pressing and sonication. It also preserves the activities of labile enzymes and releases periplasmic enzymes. Vortexing with glass beads appears to be the simplest method for cell disruption.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.3
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• pp.284-288
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• 2005

Baker's yeast was disrupted in a 1.4-L stainless steel horizontal bead mill under a continuous recycle mode using 0.3 mm diameter zirconia beads as abrasive. A single pass in continuous mode bead mill operation liberates half of the maximally released protein. The maximum total protein release can only be achieved after passaging the cells 5 times through the disruption chamber. The degree of cell disruption was increased with the increase in feeding rate, but the total protein release was highest at the middle range of feeding rate (45 L/h). The total protein release was increased with an increase in biomass concentration from 10 to $50\%$(w/v). However, higher heat dissipation as a result of high viscosity of concentrated biomass led to the denaturation of labile protein such as glucose 6-phosphate dehydrogenase (G6PDH). As a result the highest specific activity of G6PDH was achieved at biomass concentration of $20\%$(ww/v). Generally, the degree of cell disruption and total protein released were increased with an increase in impeller tip speed, but the specific activity of G6PDH was decreased substantially at higher impeller tip speed (14 m/s). Both the degree of cell disruption and total protein release increased, as the bead loading increased from 75 to $85\% (v/v)$. Hence, in order to obtain a higher yield of labile protein such as G6PDH, the yeast cell should not be disrupted at biomass concentration and impeller tip speed higher than $20\%(w/v)$ and 10 m/s, respectively.

• Korean Journal of Food Science and Technology
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• v.53 no.6
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• pp.819-822
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• 2021

To enhance phage endolysin-mediated cell wall disruption of Escherichia coli O157:H7, the cells were co-treated with aromatic compounds, namely thymol or eugenol, found in essential oils and endolysin LysECP26. Interestingly, the minimal inhibitory concentrations of LysECP26 was four times lower when used in combination with either of the two compounds than when it was used alone. This synergistic activity was also confirmed by viable cell counting. Within 1 h of LysECP26 and eugenol or thymol co-treatment to the cells, there was a 2.3 or 3.8 log CFU/mL reductions, respectively. Additionally, field emission scanning electron microscopy showed cell wall disruption and severe morphological alterations of the cells in case of the combination treatments. Therefore, endolysin and thymol or eugenol co-treatment can help in developing efficient bio-control strategies against gram-negative pathogen E. coli O157:H7.

• Mycobiology
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• v.38 no.2
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• pp.102-107
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• 2010

We identified a gene for $\beta$-1,3-glucan synthesis (GBG1), a nonessential gene whose disruption alters cell wall synthesis enzyme activities and cell wall composition. This gene was cloned by functional complementation of defects in $\beta$-1,3-glucan synthase activity of the the previously isolated Saccharomyces cerevisiae mutant LP0353, which displays a number of cell wall defects at restrictive temperature. Disruption of the GBG1 gene did not affect cell viability or growth rate, but did cause alterations in cell wall synthesis enzyme activities: reduction of $\beta$-1,3-glucan synthase and chitin synthase III activities as well as increased chitin synthase I and II activities. GBG1 disruption also showed altered cell wall composition as well as susceptibility toward cell wall inhibitors such as Zymolyase, Calcofluor white, and Nikkomycin Z. These results indicate that GBG1 plays a role in cell wall biogenesis in S. cerevisiae.

• BMB Reports
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• v.52 no.2
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• pp.111-112
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• 2019

Although many studies have reported that the breakdown of the blood-brain barrier (BBB) represents one of the major pathological changes in aging, the mechanism underlying this process remains relatively unexplored. In this study, we described that acid sphingomyelinase (ASM) derived from endothelial cells plays a critical role in BBB disruption in aging. ASM levels were elevated in the brain endothelium and plasma of aged humans and mice, resulting in BBB leakage through an increase in caveolae-mediated transcytosis. Moreover, ASM caused damage to the caveolae-cytoskeleton via protein phosphatase 1-mediated ezrin/radixin/moesin dephosphorylation in primary mouse brain endothelial cells. Mice overexpressing brain endothelial cell-specific ASM exhibited acceleration of BBB impairment and neuronal dysfunction. However, genetic inhibition and endothelial specific knock-down of ASM in mice improved BBB disruption and neurocognitive impairment during aging. Results of this study revealed a novel role of ASM in the regulation of BBB integrity and neuronal function in aging, thus highlighting the potential of ASM as a new therapeutic target for anti-aging.

• Journal of the Korean Society of Manufacturing Process Engineers
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• v.20 no.10
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• pp.63-71
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• 2021

Using fossil fuels in existing industrial systems causes a variety of social problems. Recently, many studies have been conducted on bio-refineries, which aim to actively utilize biomass to reduce the use of fossil fuels and solve various social problems. Among them, research using microalgae as a third-generation biomass has attracted considerable attention. Microalgae use inorganic matter to produce organic matter, and cell destruction is necessary to extract useful organic materials from microalgae. The extracted organic materials are currently used in various industrial fields. Numerous cell-destruction methods exist. We have investigated cell disruption by sonication, especially its efficiency. Ultrasound is a sound wave with frequencies above 20 kHz, and destroys cells by sending high energy through a cavitation that occurs, according to the characteristics of the sound wave. The Dunaliella salina microalgae used in this study was cultured in a flat-type photobioreactor. Experiments were performed using a batch low-frequency processing device. Logistic model was applied to analyze the results of cell-destruction experiments using ultrasound. The proper conditions for the most efficient cell destruction were OD 1.4(microalgae concentration)), 54watt(output power) and 200mL(microalgae capacity).

• IMMUNE NETWORK
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• v.23 no.3
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• pp.29.1-29.23
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• 2023

Cholesterol (CL) is required for various biomolecular production processes, including those of cell membrane components. Therefore, to meet these needs, CL is converted into various derivatives. Among these derivatives is cholesterol sulfate (CS), a naturally produced CL derivative by the sulfotransferase family 2B1 (SULT2B1), which is widely present in human plasma. CS is involved in cell membrane stabilization, blood clotting, keratinocyte differentiation, and TCR nanocluster deformation. This study shows that treatment of T cells with CS resulted in the decreased surface expression of some surface T-cell proteins and reduced IL-2 release. Furthermore, T cells treated with CS significantly reduced lipid raft contents and membrane CLs. Surprisingly, using the electron microscope, we also observed that CS led to the disruption of T-cell microvilli, releasing small microvilli particles containing TCRs and other microvillar proteins. However, in vivo, T cells with CS showed aberrant migration to high endothelial venules and limited infiltrating splenic T-cell zones compared with the untreated T cells. Additionally, we observed significant alleviation of atopic dermatitis in mice injected with CS in the animal model. Based on these results, we conclude that CS is an immunosuppressive natural lipid that impairs TCR signaling by disrupting microvillar function in T cells, suggesting its usefulness as a therapeutic agent for alleviating T-cell-mediated hypersensitivity and a potential target for treating autoimmune diseases.