• KSBB Journal
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• v.18 no.2
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• pp.155-160
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• 2003

The effect of dissolved oxygen(DO) on microbial transglutaminase(mTG) production by Streptoverticillium morbaraense was studied in on-line computer controlled fermentation system. In order to control dissolved oxygen during fermentation, the agitation speed and aeration rate of 2.5 L fermenter ranged from 260 to 360 rpm and 0.3 to 3.9 L/min, respectively. The maximum microbial transglutaminase production was obtained at controlled 20% of dissolved oxygen among the various dissolved oxygen controlled batch cultures tested. The production of microbial transglutaminase at controlled 20% of dissolved oxygen was about 2.12 U/mL which was 1.1 times higher than that obtained in batch culture without control of dissolved oxygen. Also, the highest microbial transglutaminase production was obtained in fed-batch cultures in which dissolved oxygen was controlled at 20%, and it was improved almost 1.3 times in comparison with that without control of dissolved oxygen. Maximal dry cell weight and microbial transglutaminase production were 13.2 g/L and 2.6 U/mL, respectively. Finally, it was also found that fed-batch fermentation at controlled 20% of dissolved oxygen showed a good performance for the microbial transglutaminase production by on-line computer controlled fermentation system which may be generally applicable to other microbial cultures.

• Journal of Microbiology and Biotechnology
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• v.10 no.2
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• pp.187-194
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• 2000

An actinomycetes strain, T-2, which produces transglutaminase (EC 2.3.2.13), was isolated from soil and identified as belonging to the Actinomadura sp., based on taxonomc studies. The conditions for the transglutaminase production and its enzymatic properties were investigated. The optimum components for the transglutaminase production were 2% glucose, 1% polypeptone and soytone, and 0.1% MnCl2. The optimum pH and temperature of the enzyme reaction were pH 8.0 and $45^{\circ}C$, respectively. The enzyme was stable within the pH range of 5.0-9.0 and $30^{\circ}C-45^{\circ}C$. The novel enzyme required no calcium ions for its activity. This enzyme polymerized various proteins such as casien, soy protein, hemoglobin, egg white, gelatin, and soybean milk.

• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.6
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• pp.801-806
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• 2009

A species producing transglutaminase (EC 2.3.2.13) was isolated from forest soil and identified as Streptomyces platensis YK-2. The transglutaminase was purified from culture broth by 50% methanol precipitation, followed by successive chromatography on DEAE-Sephadex. The yield and purification-fold was 63.4% and 2.2-fold, respectively. The purified microbial transglutaminase (MTG) migrated as a single band of approximately 45 kDa upon sodium dodecyl sulfate polyacrylamide gel eletrophoresis. The isoelectric point determined by multichambered electrofocusing was pH $6.0{\sim}7.0$. The enzyme was strongly inhibited by $Hg^{++}$, but was activated by $Cd^{++}$, $Mg^{++}$, $Mn^{++}$, $Pb^{++}$ and reducing agents such as dithiothreitol and mercaptoethanol.

• Molecules and Cells
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• v.27 no.3
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• pp.337-343
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• 2009

We developed a novel on-chip activity assay using protein arrays for quantitative and rapid analysis of transglutaminase activity in mammalian cells. Transglutaminases are a family of $Ca^{2+}$-dependent enzymes involved in cell regulation as well as human diseases such as neurodegenerative disorders, inflammatory diseases and tumor progression. We fabricated the protein arrays by immobilizing N,N'-dimethylcasein (a substrate) on the amine surface of the arrays. We initiated transamidating reaction on the protein arrays and determined the transglutaminase activity by analyzing the fluorescence intensity of biotinylated casein. The on-chip transglutaminase activity assay was proved to be much more sensitive than the $[^3H]putrescine$-incorporation assay. We successfully applied the on-chip assay to a rapid and quantitative analysis of the transglutaminase activity in all-trans retinoic acid-treated NIH 3T3 and SH-SY5Y cells. In addition, the on-chip transglutaminase activity assay was sufficiently sensitive to determine the transglutaminase activity in eleven mammalian cell lines. Thus, this novel on-chip transglutaminase activity assay was confirmed to be a sensitive and high-throughput approach to investigating the roles of transglutaminase in cellular signaling, and, moreover, it is likely to have a strong potential for monitoring human diseases.

• Food Science and Biotechnology
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• v.16 no.2
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• pp.223-227
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• 2007

Ground garlic inhibited the cross-linking reaction of myosin and incorporation of monodansylcadaverine (MDC) in salted Alaska pollack surimi catalyzed by transglutaminase (TGase). The component responsible for the inhibition was a thermostable, low molecular weight compound. The component also inhibited microbial transglutaminase (MTGase). The inhibition by garlic was reversibly recovered upon addition of 2-mercaptoethanol. The inhibitory component was therefore hypothesized to contain sulfhydryl groups within its structure. Alliin itself did not inhibit the cross-linking reaction. However, the addition of alliin together with garlic increased the inhibition. This result suggested that compounds derived from alliin was responsible for the inhibition of TGase activity.

• Journal of Microbiology
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• v.41 no.2
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• pp.161-164
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• 2003

Recombinant light chain of Clostridium botulinum neurotoxin type B stimulated transglutaminase activity in a dose dependent manner, Compared to native toxin, recombinant light chain showed av greater stimulatory effect on transglutaminase activity. Zn-chelating agents, inhibiting the proteolytic activity of the clostridial toxins, did not interfere with this stimulation. These results suggest that the light chain plays a major stimulatory role, which is not due to its metallopeptidase activity, but is possibly due to specific interaction with transglutaminase. More importantly, this report provides a new insight into the intracellular action of C. botulinum neurotoxins.

• The Journal of Korean Medicine
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• v.25 no.4
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• pp.188-199
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• 2004

Transglutaminase 2 (TGase 2) is an enzyme that is widely used in many biological systems for generic tissue stabilization purposes or immediate defenses for wounds. Many reports have showed that TGase 2 is aberrantly activated in tissues and cells and contributes to a variety of diseases, including neurodegenerative diseases and autoimmune diseases. In most cases, the TGase 2 appears to be a factor in the formation of inappropriate proteinaceous aggregates that may be cytotoxic. However, in other cases such as celiac disease, arthritis, lupus, amyotrophic lateral sclerosis, TGase 2 is involved in the generation of autoantibodies. This suggests the possibility that the inappropriate expression and/or presentation of TGase 2 to T cells might contribute to these diseases in genetically predisposed individuals. Others and we have found that TGase 2 expression is also increased in the inflammation process. We also demonstrated reverse of inflammation by TGase inhibition. Furthermore we discovered the genuine role of TGase 2 in immune cell activation. Increase of TGase activity induces or exacerbates inflammation via NF-κB activation without I-κBα kinase signalings. This review will examine a possibility of TGase inhibitors as therapeutic agents in a variety of inflammatory diseases.

• Journal of Microbiology and Biotechnology
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• v.19 no.6
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• pp.588-595
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• 2009

A bacterial strain, YK-2, was isolated as a producer of trans glutaminase from a forest soil sample of Daegu, Korea. The isolate showed a G+C content of 72.7 mol%, contained meso-$A_2pm$ as the cell-wall amino acid, and possessed menaquinone MK-9 ($H_6$) and menaquinone MK-9 ($H_8$) at a ratio of 6:4. The chemotaxonomic analysis, as well as phylogenetic analysis based on the 16S rDNA sequence, identified the isolate as a member of Streptomyces platensis. For transglutaminase production, the optimum medium composition was determined to be 2% glucose, 1% polypeptone, 1% soy tone, and 0.1% $MnCl_2$. The transglutaminase was stable within the pH range of 5.0-9.0 and $30-45^{\circ}C$, and the optimum pH and temperature were pH 8.0 and $45^{\circ}C$, respectively, without any requirement for $Ca^{2+}$.

• Food Science and Biotechnology
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• v.17 no.5
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• pp.904-911
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• 2008

Covalent cross-links between a number of proteins and peptides explain why transglutaminase may be widely used by food processing industries. The objective of this work was optimization of the fermentation process to produce transglutaminase from a new microbial source, the Streptomyces sp. P20. The strategy adopted to modify the usual literature media was: (1) fractional factorial design (FFD) to elucidate the key medium ingredients, (2) central composite design (CCD) to optimise the concentration of the key components. Optimization of the medium resulted in not only an 86% increase in microbial transglutaminase activity as compared to the media cited in the literature, but also a reduction in the production cost. Optimal fermentation conditions - namely temperature and agitation rate - were also studied, using CCD methodology. Usual conditions of $30^{\circ}C$ and 100 rpm were within the optimal area. All other parameters for enzyme production were experimentally proven to be optimum fermentation conditions.

• The Korean Journal of Mycology
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• v.36 no.1
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• pp.93-97
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• 2008

A $Ca^{2+}-independent$ microbial transglutaminase (mTGase) from the actinomycete Streptomyces mobaraensis IFO13819 is a useful enzyme in the food industry. It is consists 406 amino acid residues, which comprised leader and pro region of 75 amino acid residues and the structure region of 331 amino acid residues. Pro and structure gene of TGase were cloned into the yeast shuttle vector pYAEG-TER and then used to transform Saccharomyces cerevisiae 2805. Expression of mTGase in recombinant was confirmed with Northern hybridization and the maximal activity of TGase was shown 26 mU/ml.