• Title/Summary/Keyword: Enzyme I

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Panax ginseng total protein promotes proliferation and secretion of collagen in NIH/3T3 cells by activating extracellular signal-related kinase pathway

  • Chen, Xuenan;Wang, Manying;Xu, Xiaohao;Liu, Jianzeng;Mei, Bing;Fu, Pingping;Zhao, Daqing;Sun, Liwei
    • Journal of Ginseng Research
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    • v.41 no.3
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    • pp.411-418
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    • 2017
  • Background: Recently, protein from ginseng was studied and used for the treatment of several kinds of diseases. However, the effect of ginseng total protein (GTP) on proliferation and wound healing in fibroblast cells remains unclear. Methods: In this study, cell viability was analyzed using the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. Cell cycle distribution was analyzed by flow cytometer. The levels of transforming growth factor ${\beta}1$, vascular endothelial growth factor, and collagens were analyzed by enzyme-linked immunosorbent assay and immunofluorescence staining. The expressions of cyclin A, phosphorylation of extracellular signal-related kinase (p-ERK1/2), and ERK1/2 were analyzed by Western blotting. Results: Our results showed that GTP promoted cell proliferation and increased the percentage of cells in S phase through the upregulation of cyclin A in NIH/3T3 cells. We also found that GTP induced the secretion of type I collagen, and promoted the expression of other factors that regulate the synthesis of collagen such as transforming growth factor ${\beta}1$ and vascular endothelial growth factor. In addition, the phosphorylation of ERK1/2 at Thr202/Tyr204 was also increased by GTP. Conclusion: Our studies suggest that GTP promoted proliferation and secretion of collagen in NIH/3T3 cells by activating the ERK signal pathway, which shed light on a potential function of GTP in promoting wound healing.

Compatibility effects of ginseng and Ligustrum lucidum Ait herb pair on hematopoietic recovery in mice with cyclophosphamide-induced myelosuppression and its material basis

  • Han, Jiahong;Dai, Min;Zhao, Yan;Cai, Enbo;Zhang, Lianxue;Jia, Xiaohuan;Sun, Nian;Fei, Xuan;Shu, Hui
    • Journal of Ginseng Research
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    • v.44 no.2
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    • pp.291-299
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    • 2020
  • Background: Ginseng (G) and Ligustrum lucidum Ait (LLA) are core traditional Chinese medicines in treating myelosuppression formula. The present study was designed to profile effect of G and LLA herb pair (G-LLA) on myelosuppressed mice. Methods: The mice myelosuppression model was established by intraperitoneal (i.p.) injection of cyclophosphamide (Cy). Hematopoietic function of bone marrow was measured by hemopoietic progenitor cell culture and peripheral blood count, and serum hemopoietic factors were tested by enzyme-linked immunosorbent assay. Bone marrow cell cycle was performed by flow cytometry. HPLC was used to measure 20 potential chemical components related to myelosuppression, including ginsenoside Rg1, Re, Rb1, Rc, Rb2, Rb3, Rd, Rk3, Rh4, 20 (S)-Rg3, 20 (R)-Rg3, Rk1, Rg5, salidroside, and so on. Results: G, LLA, and G-LLA improved the amount of peripheral blood cells and bone marrow cells of myelosuppressed mice (P < 0.01). They significantly increased the colony quantity of colony-forming unit-granulocyte macrophage, burst-forming unit-erythroid, colony-forming unit-erythroid, and colony-forming unit-megakaryocyte and amount of G2/M and S phase cells (P < 0.01). They also significantly decreased the amount of hematopoiesis-related cytokines (P < 0.01). The content of chemical components in G-LLA changed, and the change of rare saponin was the most obvious. Conclusion: These results show that G-LLA herb pair might produce synergistic or complementary compatibility effects on bone marrow suppression after chemotherapy. It suggests that the substance basis of G-LLA for treating bone marrow suppression may be effective chemical components.

Enzymatic Modification of Soy Proteins: Effects of Functional Properties of Soy Isolate upon Proteolytic Hydrolysis (대두단백질(大豆蛋白質)의 효소적(酵素的) 변형(變形) : 분리대두단백질(分離大豆蛋白質)의 기능성(機能性)에 미치는 단백질가수분해(蛋白質加水分解)의 영향(影響))

  • Kang, Yeung-Joo
    • Korean Journal of Food Science and Technology
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    • v.16 no.2
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    • pp.211-217
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    • 1984
  • To study affinity of proteolytic enzymes to soy proteins, the physicochemical and functional properties of enzymatically modified protein products, kinetic parameters and degree of hydrolysis were measured using trypsin, alcalase (serine type protease) and pronase. Bacterial alcalase and pronase showed much greater affinity to soy protein than animal intestinal trypsin. This effect was very significant when unheated soy isolate was used as a substrate. Specific activities of these enzymes decreased with the increment of substrate concentration (over 2.0%, w/v) when heat denatured soy protein was used as a substrate. However, the decrease in specific activity was negligible at substrate concentrations lower than 2.0%. Polyacrylamide gel electrophoretic results showed that the pattern of 2S protein band changed distinctly in alcalase hydrolysis as compared with those of trypsin and pronase. Protein solubilities of alcalase and pronase hydrolyzates increased by 25-30%, at their pI (pH 5.0) over the control. Virtually no change was observed in solubility by trypsin hydrolysis. Heat coagulability and calcium-tolerance of the protein increased by enzymatic hydrolysis. No clear tendency, however, was observed for emulsion properties, foam expansion and the amount of free -SH groups. The enzyme treatment considerably decreased foam stability.

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The Effects of Gardeniae Fructus Aqua-Acupuncture on Liver Injury of Rats Induced by CCI4 (치자약침(梔子藥鍼)이 실험적(實驗的) 백서(白鼠)의 간손상(肝損傷)에 미치는 영향(影響))

  • Park, Hee-Soo;Park, Joung-Hyun
    • Journal of Pharmacopuncture
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    • v.3 no.2
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    • pp.55-77
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    • 2000
  • This is the study of the effects of Aqua-acupuncture with Gardeniae Fructus on thc recovery of rat's liver which was damaged by 0.3ml/ea of $CCI_4$. Rats were divided into 4 groups; Normal-group(None treated group), Control-group(Not treated after $CCI_4$-intoxicated), Exp. I(Treated with Saline Aqua-acupuncture after $CCI_4$-intoxicated) and Exp. ll(Treated with Gardeniae Fructus Aqua-acupuncture after $CCI_4$-intoxicated). Biochemical assays for each serum enzyme activities of AST, ALT, Albumin, LDH, ${\gamma}$-GT, TG and Total cholesterol were performed. The results were summarized as follows: 1. AST activities in serum significantly decreased in the Gardeniae Fructus Aqua-acupuncture treated group after $CCI_4$-intoxicated. In companson with Saline-treated group after $CCI_4$-intoxicated, the Gardeniae Fructus Aqua-acupuncture treated group *The professor of Dept. of Acupuncture & Moxibustion, 2. At T activities in serum significantly decreased in the Gardeniae Fructus Aqua-acupuncture treated group after $CCI_4$-intoxicated. In com pan son with Saline-treated group after $CCI_4$-intoxicated, the Gardeniae Fructus Aqua-acupuncture treated group after $CCI_4$-intoxicated worked effectively to rat's damaged liver. 3. Albumin in serum increased in the Gardeniae Fructus Aqua- acupurkture treated group after $CCI_4$-intoxicated. 4. LDH in serum significantly decreased in the Gardeniae Fructus Aqua-acupuncture treated group after $CCI_4$-intoxicated. In comparison with Saline-treated group after CClcintox icated, the Gardeniae Fructus Aqua acupuncture treated group after $CCI_4$-intoxicated worked highly effectively to rat's damaged liver. 5. ${\gamma}$-GT In serum significantly decreased In the Gardeniae Fructus Aqua-acupuncture trea ted group after $CCI_4$-intoxicated. In compan son with Saline-treated group after $CCI_4$-intoxicated, the Crardeniae Fructus Aqua-acupuncture treated group after $CCI_4$-intoxicated was not recognized significantly. 6. TG in serum significantly decreased in the Gardeniae Fructus Aqua-acupuncture treated group after $CCI_4$-intoxicated. In comparison with Saline-treated group after $CCI_4$-intoxicated, the Gardeniae Fructus Aqua -acupuncture treated group after $CCI_4$-intoxicated worked highly effectively to rat's damaged liver. 7. Total cholesterol in serum decreased in the Gardenias Fructus Aqua-acupuncture treated group after $CCI_4$-intoxicated. In comparison with Saline-treated group after $CCI_4$-intoxicated, the Gardeniae Fructus Aqua-acupuncture treated group after $CCI_4$-intoxicated worked highly effectively to rat's damaged liver. The results from ahove show that the Gardeniae Fructus Aqua- acupuncture has highly effects on the damaged liver caused by $CCI_4$. Therefore it is expected that the Gardeniae Fructus Aqua- acupuncture could be used to cure the damaged liver.

Cardiovascular Biofunctional Activity and Antioxidant Activity of Gugija (Lycium chinensis Mill) Species and Its Hybrids (구기자 품종과 교배종의 부위별 심혈관관련 질환 예방 기능성 및 항산화 활성비교)

  • Park, Won-Jong;Lee, Bong-Chun;Lee, Ju-Chan;Lee, Eun-Na;Song, Jung-Eun;Lee, Dae-Hyung;Lee, Jong-Soo
    • Korean Journal of Medicinal Crop Science
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    • v.15 no.6
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    • pp.391-397
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    • 2007
  • To develop new high valuable Gugija (Lycium chinensis), biofunctionalities of Gugija standard species and its hybrids were investigated and compared with each water extracts and methanol extracts from Lycii Fructus, Lycii Folium and buds and Lycii Cortex Radicis. Among various biofunctionalities of Gugija standard species, antioxidant activity was showed the highest in methanol extracts from buds of Cheongwoon species (93%) and antihypertensive angiotensin I-converting enzyme (ACE) inhibitory activity was 84.1% in the water extracts from Lycii Cortex Radicis of Cheongyang NO.7. Futhermore, methanol extracts from Lycii Cortex Radicis of Myungan A-2 hybrid showed 93.1% of antioxidant activity and 96.9% of ACE inhibitory activity was also showed in the methanol extracts from Lycii Fructus of DO148-72(A11) hybrid. However, fibrinolytic activity and anticholesteromia HMG-CoA reductase inhibitory activity were weak or not detected in almost of Gugija standard species and its hybrids. Therefore, we finally selected Cheongwoon Gugija standard species (buds) and Myungan A-2 hybrid (Lycii Cortex Radicis) as good antioxidant sources and also DO148-72 (A11) hybrid (Lycii Fructus) as excellent antihypertensive ACE inhibitior sources for manufacturing functional food product.

L-Lysine Production by 6-Azauracil Resistant Mutant of Corynebacterium glutamicum (6-Azaumcil 내성을 지닌 Corynebacterium glutamicum 변이주에 의한 L-Lysine의 생산)

  • 신현철;김성준전영중이재흥
    • KSBB Journal
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    • v.9 no.4
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    • pp.372-377
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    • 1994
  • To improve L-lysine yield, pyrimidine base analogue(6-azauracil)-resistant mutants were isolated from Corynebacterium glutamicum KFCC10672 Among them the best producer, C. glutamicum CH0516, was selected and tested for L-lysine production in a $7\ell$ fermentor. It was found that the product yield obtained with C. glutamicum CH0516 was higher than that of the parent strain by 3%. In order to elucidate the gain in productivity with the 6-azauracil-resistant mutant enzymatic kinetic parameters such as aspartokinase(AKase) and aspartate carbamoyltransferase (ATCase) were measured. The Km values of AKase with C. glutamicum KFCC10672 and CH0516 were 200.0 mM and 166.7 mM and those of ATCase were 0.13 mM and 0.27 mM, respectively. However, the specific enzyme activities of AKase of C. glutamlcum KFCC10672 and CH0516 were $3.89{\times}10^{-1}$ units/mg and $4.78{\times}10^{-1}$ units/mg, and those of ATCarse were 2.20 units/mg and 1.84 units/mg, respectively. It appears that some increase in product yield with C. gluramicum CH0516 is likely due to the increased Akase activity and decreased ATCase activity.

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Physicochemical and Functional Properties of Kochujang with Broccoli Leaf Powder (브로콜리잎 분말 첨가 재래식 고추장의 이화학적 및 기능적 특성)

  • Oh, You-Sung;Baek, Jin-Woo;Park, Kyeong-Yeol;Hwang, Joon-Ho;Lim, Sang-Bin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.5
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    • pp.675-681
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    • 2013
  • Home-made broccoli Kochujang (HMBK) was prepared with the addition of 5% broccoli leaf powder. Its physicochemical and functional properties were measured in extracts (80% methanol, 80% ethanol, and distilled water) and compared with home-made Kochujang (HMK) and factory-produced Kochujang (FPK). Total phenolic content (TPC) was 22% higher in methanol extract from HMBK (524.2 GAE/100 g) compared to HMK (431.0 GAE/100 g). TPC was 8% higher in ethanol extract from HMBK (541.9 GAE/100 g) compared to HMK (499.9 GAE/100 g). DPPH radical scavenging activity was 1.6 times higher in the methanol extracts from HMBK than HMK. In contrast there was no difference in DPPH radical scavenging activity between HMBK and HMK. Oxygen radical absorbance capacities in methanol and ethanol extracts from HMBK were similar to HMK, but both were higher than extracts from FPK (55% and 23% higher, respectively). Inhibition of angiotensin I converting enzyme activity in methanol extracts from HMBK was similar to HMK, but it was 2.6 times higher than inhibition activities from FPK. Interestingly, only ethanol extract from HMBK showed a 10.7% and 18.3% inhibition on cell growth of the human colon adenocarcinoma grade II cell line (HT-29) and human lung carcinoma cell line (NCI-H1229), respectively. These results indicate home-made Kochujang with broccoli leaf powder contains high total phenolics, antioxidant activities, and cancer cell growth inhibition activities.

The Hepatoprotective Effects of Epimedii Herba through the Antioxidation (음양곽의 항산화작용에 의한 간 보호 효과)

  • Ha Bae Jin;Kim Hee Jin;Lee Sang Hun;Ha Jong-Myung;Lee Sang-Hyeon;Lee Jae-Hwa;Lee Dong-Geun;Park Eun Kyung;Nam Chun Suk
    • Journal of Life Science
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    • v.15 no.4 s.71
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    • pp.572-577
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    • 2005
  • In this study, the effect of Epimedii Herba (EH) on the antioxidative enzymatic activity was investigated. EH (100mg/kg) was intraperitoneally administered into rats for 2 weeks. On the last day, carbon tetrachloride $(50\%\;CCl_4,\;3.3ml/kg,\;i.p.)$ dissolved in olive oil was injected before 12 hours. EH-pread-ministered and $CCl_4-treated$ (EC) group showed higher inhibitory effect in aminotransferase (AST, ALT) activity compared to $CCl_4-treated (CT)$ group. Superoxide dismutase (SOD) and Catalase in EC group were increased compared to those of CT group. The activity of glutathione peroxidase (GPx) was significantly higher than those of CT group compared to EC group. These results suggest that EH has a hepatoprotective effect through scavenging the free radicals induced by $CCl_4$.

Synthesis of L-threo-3,4-Dihydroxyphenylserine(L-threo-DOPS) with Thermostabilized Low-Specific L-Threonine Aldolase from Streptomyces coelicolor A3(2)

  • Baik, Sang-Ho;Yoshioka, Hideki;Yukawa, Hideaki;Harayama, Shigeaki
    • Journal of Microbiology and Biotechnology
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    • v.17 no.5
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    • pp.721-727
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    • 2007
  • Stability-enhanced mutants, H44, 11-94, 5A2-84, and F8, of L-threonine aldolase(L-TA) from Streptomyces coelicolor A3(2)(SCO1085) were isolated by an error-prone PCR followed by a high-throughput screening. Each of these mutant, had a single amino acid substitution: H177Y in the H44 mutant, A169T in the 11-94 mutant, D104N in the 5A2-84 mutant and F18I in the F8 mutant. The residual L-TA activity of the wild-type L-TA after a heat treatment for 20 min at $60^{\circ}C$ was only 10.6%. However, those in the stability-enhanced mutants were 85.7% for the H44 mutant, 58.6% for the F8 mutant, 62.1% for the 5A2-84 mutant, and 67.6% for the 11-94 mutant. Although the half-life of the wild-type L-TA at $63^{\circ}C$ was 1.3 min, those of the mutant L-TAs were longer: 14.6 min for the H44 mutant, 3.7 min for the 11-94 mutant, 5.8 min for the 5A2-84 mutant, and 5.0 min for the F8 mutant. The specific activity did not change in most of the mutants, but it was decreased by 45% in the case of mutant F8. When the aldol condensation of glycine and 3,4-dihydroxybenzaldehyde was studied by using whole cells of Escherichia coli containing the wild-type L-TA gene, L-threo-3,4-dihydroxyphenylserine(L-threo-DOPS) was successfully synthesized with a yield of 2.0 mg/ml after 20 repeated batch reactions for 100 h. However, the L-threo-DOPS synthesizing activity of the enzyme decreased with increased cycles of the batch reactions. Compared with the wild-type L-TA, H44 L-TA kept its L-threo-DOPS synthesizing activity almost constant during the 20 repeated batch reactions for 100 h, yielding 4.0 mg/ml of L-threo-DOPS. This result showed that H44 L-TA is more effective than the wild-type L-TA for the mass production of L-threo-DOPS.

A Newly Identified Glutaminase-Free L-Asparaginase (L-ASPG86) from the Marine Bacterium Mesoflavibacter zeaxanthinifaciens

  • Lee, Su-Jin;Lee, Youngdeuk;Park, Gun-Hoo;Umasuthan, Navaneethaiyer;Heo, Soo-Jin;Zoysa, Mahanama De;Jung, Won-Kyo;Lee, Dae-Won;Kim, Hanjun;Kang, Do-Hyung;Oh, Chulhong
    • Journal of Microbiology and Biotechnology
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    • v.26 no.6
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    • pp.1115-1123
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    • 2016
  • L-Asparaginase (E.C. 3.5.1.1) is an enzyme involved in asparagine hydrolysis and has the potential to effect leukemic cells and various other cancer cells. We identified the L-asparaginase gene (L-ASPG86) in the genus Mesoflavibacter, which consists of a 1,035 bp open reading frame encoding 344 amino acids. Following phylogenetic analysis, the deduced amino acid sequence of L-ASPG86 (L-ASPG86) was grouped as a type I asparaginase with respective homologs in Escherichia coli and Yersinia pseudotuberculosis. The L-ASPG86 gene was cloned into the pET-16b vector to express the respective protein in E. coli BL21 (DE3) cells. Recombinant L-asparaginase (r-L-ASPG86) showed optimum conditions at 37-40℃, pH 9. Moreover, r-L-ASPG86 did not exhibit glutaminase activity. In the metal ions test, its enzymatic activity was highly improved upon addition of 5 mM manganese (3.97-fold) and magnesium (3.35-fold) compared with the untreated control. The specific activity of r-L-ASPG86 was 687.1 units/mg under optimum conditions (37℃, pH 9, and 5 mM MnSO4).