• Title/Summary/Keyword: TGase

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pH 조정후 Transglutaminase로 처리한 원유의 전자현미경적 관찰

  • Mun, Jeong-Han;Hong, Yun-Ho
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 2004.10a
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    • pp.380-383
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    • 2004
  • 본 연구는 원유에서 지방을 제거한 탈지유의 pH를 5.5, 7.0, 8.5로 조정한 다음 TGase를 첨가하여 0, 1, 2, 4, 8시간 반응시킨 다음 단백질 입자들을 동결건조하여 조직의 성상에 대해 주사 전자 현미경을 이용해 관찰, 비교하였다. pH와 TGase를 처리하지 않은 원유의 탈지유는 단백질 입자들이 규칙적으로 회합해 있었다. 그러나 pH 조정 후 TGase를 처리한 다음 반응시간을 달리한 시료에서는 pH를 5.5로 조정한 시료에서 현저한 변화가 있었는데 그 변화 양상은 단백질 입자들이 0시간에서 조각을 이루워 회합되어 있다가 1시간 반응시킨 경우 단백질 입자들이 서로 결합하여 넓게 회합을 하였다. 2시간 반응시킨 경우 단백질 입자들이 다시 뭉쳐서 회합하였으며 4시간 반응시킨 경우 뭉쳐져 있던 단백질 입자들이 조그만한 구형 성상으로 넓게 회합하였다. 8시간 반응시킨 시료는 구형 성상으로 회합되어 있던 단백질 입자들이 사라지면서 다시 넓게 회합하는 것을 관찰할 수 있었다. pH 7.0과 8.5 조건하에서는 단백질 입자들이 조각 형태를 이루고 있었으며 반응시간이 증가할수록 입자들이 넓게 확대되는 현상을 나타냈다. 이와 같은 단백질들의 변화 양상은 pH와 TGase처리 그리고 반응시간에 영향을 받고 있는 것으로 사료된다.

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Ethacrynic Acid Inhibits Sphingosylphosphorylcholine-Induced Keratin 8 Phosphorylation and Reorganization via Transglutaminase-2 Inhibition

  • Byun, Hyun Jung;Kang, Kyung Jin;Park, Mi Kyung;Lee, Hye Ja;Kang, June Hee;Lee, Eun Ji;Kim, You Ri;Kim, Hyun Ji;Kim, Young Woo;Jung, Kyung Chae;Kim, Soo Youl;Lee, Chang Hoon
    • Biomolecules & Therapeutics
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    • v.21 no.5
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    • pp.338-342
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    • 2013
  • Sphingosylphosphorylcholine (SPC) is significantly increased in the malicious ascites of tumor patients and induces perinuclear reorganization of keratin 8 (K8) filaments in PANC-1 cells. The reorganization contributes to the viscoelasticity of metastatic cancer cells resulting in increased migration. Recently, we reported that transglutaminase-2 (Tgase-2) is involved in SPC-induced K8 phosphorylation and reorganization. However, effects of Tgase-2 inhibitors on SPC-induced K8 phosphorylation and reorganization were not clearly studied. We found that ethacrynic acid (ECA) concentration-dependently inhibited Tgase-2. Therefore, we examined the effects of ECA on SPC-induced K8 phosphorylation and reorganization. ECA concentration-dependently suppressed the SPC-induced phosphorylation and perinuclear reorganization of K8. ECA also suppressed the SPC-induced migration and invasion. SPC induced JNK activation through Tgase-2 expression and ECA suppressed the activation and expression of JNK in PANC-1 cells. These results suggested that ECA might be useful to control Tgase-2 dependent metastasis of cancer cells such as pancreatic cancer and lung cancers.

Gelation of Rapeseed Protein Induced with Microbial Transglutaminase (미생물성 Transglutaminase에 의한 유채단백질의 겔화)

  • Hyun, Eun-Hee;Kang, Yeung-Joo
    • Korean Journal of Food Science and Technology
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    • v.31 no.5
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    • pp.1262-1267
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    • 1999
  • Optimum reaction conditions for gel formation of rapeseed, Brassica napus, protein catalyzed by microbial TGase(transglutaminase) were evaluated by measuring breaking strength and deformation of gel. The polymerization of the protein gel was ascertained by SDS-PAGE and content of GL crosslinking$[{\varepsilon}-({\gamma}-glutamyl)lysine]$. In the reaction between rapeseed protein and TGase at $45^{\circ}C$ for 60 min, the breaking strength and deformation of the gel was the maximum at the ratio of 1 : 40 of enzyme to substrate. 10%(w/v) of rapeseed protein concentrate was optimum for gel production. The maximum breaking strength and deformation was shown at $45^{\circ}C$ The breaking strength increased linearly up to 90 min of the reaction time and remained unchanged. The breaking strength and deformation by TGase treatment was pH dependent and pH 7 was optimum for 10% rapeseed protein solution. SDS-PAGE analysis indicated that new band of highmolecular polymers were formed by the enzyme reaction, with disappearing the original bands of rapeseed protein. According to HPLC analysis. the content of GL crosslinking was increased from 0 to $7.14\;{\mu}mol/g$ gel for 90 min of the reaction time.

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Effect of Red Bean Protein and Microbial Transglutaminase on Gelling Properties of Myofibrillar Protein (적소두단백질(Red Bean Protein)과 Transglutaminase를 첨가한 돈육 근원섬유 단백질의 물성 증진 효과)

  • Jang, Ho-Sik;Chin, Koo-Bok
    • Food Science of Animal Resources
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    • v.31 no.5
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    • pp.782-790
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    • 2011
  • The effects of soy protein isolate (SPI) and red bean protein isolate (RBPI) on gelling properties of pork myofibrillar protein (MP) in the presence of microbial transglutaminase (MTG) were studied at 0.45 M NaCl. MP paste was incubated with MTG (0.1%) at various levels (0.1, 0.3, 0.5, and 1%) of SPI and RBPI before incubating at $4^{\circ}C$ for 4 h. The rheological property results showed that MP gel shear stress increased with increasing RBPI concentration. Cooking yield (CY) of the MP gel increased with increasing RBPI and SPI, whereas gel strength (GS) was not affected by adding RBPI or SPI. Thus, effects of incubation time (0, 4, 8, 10, and 12 h) were measured at 0.1% SPI and RBPI. GS values of the MP gel at 10 and 12 h were similar and were higher than those of the others. CY values were highest when RBPI (0.1%) was added, regardless of incubation time. The protein patterns indicated that incubating the MP with MTG for 10 h resulted in protein crosslinking between MP and RBPI or SPI. Based on these results, RBPI and SPI could be used as an ingredient to increase textural properties and cooking yield of meat protein gel.

Involvement of Transglutaminase-2 in α-MSH-Induced Melanogenesis in SK-MEL-2 Human Melanoma Cells

  • Kim, Hyun Ji;Lee, Hye Ja;Park, Mi Kyung;Gang, Kyung Jin;Byun, Hyun Jung;Park, Jeong Ho;Kim, Mi Kyung;Kim, Soo Youl;Lee, Chang Hoon
    • Biomolecules & Therapeutics
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    • v.22 no.3
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    • pp.207-212
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    • 2014
  • Skin hyperpigmentation is one of the most common skin disorders caused by abnormal melanogenesis. The mechanism and key factors at play are not fully understood. Previous reports have indicated that cystamine (CTM) inhibits melanin synthesis, though its molecular mechanism in melanogenesis remains unclear. In the present study, we investigated the effect of CTM on melanin production using ELISA reader and the expression of proteins involved in melanogenesis by Western blotting, and examined the involvement of transglutaminase-2 (Tgase-2) in SK-MEL-2 human melanoma cells by gene silencing. In the results, CTM dose-dependently suppressed melanin production and dendrite extension in a-MSH-induced melanogenesis of SK-MEL-2 human melanoma cells. CTM also suppressed a-MSH-induced chemotactic migration as well as the expressions of melanogenesis factors TRP-1, TRP-2 and MITF in a-MSH-treated SK-MEL-2 cells. Meanwhile, gene silencing of Tgase-2 suppressed dendrite extension and the expressions of TRP-1 and TRP-2 in a-MSH-treated SK-MEL-2 cells. Overall, these findings suggested that CTM suppresses a-MSH-induced melanogenesis via Tgase-2 inhibition and that therefore, Tgase-2 might be a new target in hyperpigmentation disorder therapy.

Effect of Time and Pressure on Quality of Restructured Pork using TGase (TGase를 사용하여 재구성돈육 제조시 결착시간과 압력조건이 품질에 미치는 영향)

  • Kim, Young-Boong;Rho, Jeong-Hae;Park, Eun-Hye
    • Journal of Animal Science and Technology
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    • v.45 no.1
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    • pp.143-150
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    • 2003
  • This study was performed to compare the effects of binding time(2hr, 4hr, 6hr and 16hr) and pressing condition (1kg, 3kg, 5kg and 10kg) on manufacturing restructured pork. Meat color, in terms of L, a, b showed no significant difference by treatment of binding time. Five kg pressing resulted in the highest red value(P<0.05). In tensile strength, 6 hour treatment showed significant difference ranging in 202.1g for raw restructured pork and 389.0g for cooked restructured pork. Five kg pressing showed the highest tensile strength, 114.6 for raw restructured pork and 303.3 for cooked restructured pork. In hardness, springiness, cohesiveness and chewiness, raw and cooked restructured pork showed insignificant difference regardless of pressing time. In sensory evaluation, four hour treated raw restructured pork showed the highest values of color, binding degree, shape, acceptance, while four or six hour treated restructured pork showed the highest values in cooked restructured pork. Therefore the optimal conditions for manufacturing restructured pork using TGase were four through six hour pressing with 5kg pressure.

Inhibition of Transglutaminase and Microbial Transglutaminase Activity by Garlic

  • Lee, Nam-Hyouck;Takeuchi, Atsuyoshi;Konno, Kunihiko
    • 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.

Effect of Maleylation on Physicochemical Properties of Soybean Glycinin

  • Shin, Weon-Sun;Park, Soo-Jin;Park, Chun-Wuk;Kim, Kang-Sung
    • Macromolecular Research
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    • v.15 no.7
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    • pp.671-675
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    • 2007
  • Soybean proteins appear to harbor a great deal of potential as functional ingredients due to the fact that they are composed of highly bioavailable peptides and amino acids. To develop drink- or gel-type foods formulated with soybean protein, the physicochemical properties of intact and chemically modified soy glycinin were assessed. Maleylation to soy glycinin altered the surface charges of glycinin via the modification of lysine residues, and subsequently generated the dissociation of glycinin subunits owing to the increase in charge repulsion. This modification thus improved the solubility of glycinin, particularly under acidic pH conditions. It is worthy of note that maleylation increased the susceptibility of the basic subunits of mTGase and the formation of a substantial quantity of molecules at a low protein solution concentration. The results of dynamic rheological studies indicated that the 5% intact glycinin progressively formed the gel with mTGase treatment in a concentration-dependent manner, but maleylated-glycinin did not.

Effects of Eucommia ulmoides Oliver Extract on Inhibition of β-hexosaminidase and Keratinocyte Differentiation (β-hexosaminidase 분비 억제 및 각질형성세포 분화에 대한 두충(Eucommia ulmoides Oliver) 추출물의 효과)

  • Hong, In Kee;Kim, Eun Ji;Seok, Ji Hyun;Kim, Bo Hyeon;Jang, Jin Dong;Joe, Gi Jung;Choi, Shin Wook
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.40 no.1
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    • pp.21-28
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    • 2014
  • In this study, Eucommia ulmoides Oliver extracts was studied in order to see any effects on the ${\beta}$-hexosaminidase release suppression of RBL-2H3 cells and on the expression of filaggrin, transglutaminase-1 (TGase-1) and cornified cell envelope (CE) related to the recovery of HaCaT keratinocyte skin barrier. Results showed that Eucommia ulmoides Oliver extracts reduced ${\beta}$-hexosaminidase release in RBL-2H3 cells and increased the effects of Eucommia ulmoides Oliver extract on the expression of filaggrin, transglutaminase-1 (TGase-1) and cornified cell envelope (CE) in HaCaT keratinocytes. Taken together, these results suggested that Eucommia ulmoides Oliver extract may be applicable for keratinocyte differentiation.