• Title/Summary/Keyword: degradation enzyme

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Isolation of Agarivorans sp. KC-1 and Characterization of Its Thermotolerant β-Agarase (한천분해세균 Agarivorans sp. KC-1의 분리 및 내열성 β-아가라제의 특성 규명)

  • Min, Kyung-Cheol;Lee, Chang-Eun;Lee, Dong-Geun;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.28 no.9
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    • pp.1056-1061
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    • 2018
  • This article reports an agar-degrading marine bacterium and characterizes its agarase. The agar-degrading marine bacterium, KC-1, was isolated from seawater on the shores of Sacheon, in Gyeongnam province, Korea, using Marine Broth 2216 agar medium. To identify the agar-degrading bacterium as Agarivorans sp. KC-1, phylogenetic analysis based on the 16S rRNA gene sequence was used. An extracellular agarase was prepared from a culture medium of Agarivorans sp. KC-1, and used for the characterization of enzyme. The relative activities at 20, 30, 40, 50, 60, and $70^{\circ}C$ were 65, 91, 96, 100, 77, and 35%, respectively. The relative activities at pH 5, 6, 7, and 8 were 93, 100, 87, and 82%, respectively. The extracellular agarase showed maximum activity (254 units/l) at pH 6.0 and $50^{\circ}C$ in 20 mM of Tris-HCl buffer. The agarase activity was maintained at 90% or more until 2 hr exposure at $20^{\circ}C$, $30^{\circ}C$ and $40^{\circ}C$, but it was found that the activity decreased sharply from $60^{\circ}C$. A zymogram analysis showed that Agarivorans sp. KC-1 produced 3 agar-degrading enzymes that had molecular weights of 130, 80, and 69 kDa. A thin layer chromatography analysis suggested that Agarivorans sp. KC-1 produced extracellular ${\beta}$-agarases as it hydrolyzed agarose to produce neoagarooligosaccharides, including neoagarohexaose (21.6%), neoagarotetraose (32.2%), and neoagarobiose (46.2%). These results suggest that Agarivorans sp. KC-1 and its thermotolerant ${\beta}$-agarase would be useful for the production of neoagarooligosaccharides that inhibit bacterial growth and delay starch degradation.

The Quality Characteristics of Rice Mash by Mixing Ratios of Rice and Rice Koji (쌀과 미입국의 배합비율에 따른 쌀 당화액의 품질특성)

  • Kim, Jin-Sook;Lee, Ji-Hyun;Chang, Young-Eun;Kim, Gi-Chang;Kim, Kyung-Mi
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.12
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    • pp.2035-2041
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    • 2013
  • The effects of Aspergillus oryzae rice koji (AO) and Asp. kawachii rice koji (AK) as enzyme preparations, on the quality characteristics of rice mash were invested in this study. The amount of AORM (Asp. oryzae rice mash) and AKRM (Asp. kawachii rice mash) were 50, 100, 200% (w/w) based on 100 g of rice. Firstly, in the titer measurement result on the ${\alpha}$-amylase and glucoamylase activities of AO and AK. On the other hand, the acid protease activity has values of 31.56 unit for AO and 849.17 unit for AK. The sugar solid of the AORM and AKRM groups significantly increased as the rice koji ratio on rice was higher, which were shown with values as high as 17.63~20.53 and 17.51~19.28, respectively. Glucose and maltose were detected for free sugar of AORM. Only glucose was found in AKRM. Citric acid, malic acid, and lactic acid were detected as the organic acid of KORM; oxalic acid, citric acid, and succinic acid were detected for AKRM, and the content increased as the rice koji ratio on rice increased (P<0.05). From the above result, rice koji with useful mold is expected to be used broadly in foods by looking at the fact that it has starch degradation ability and organic acid producibility.

Enzymatic characterization of Paenibacillus amylolyticus xylanases GH10 and GH30 for xylan hydrolysis (Paenibacillus amylolyticus 유래 xylanase GH10 및 GH30의 xylan 가수분해 특성)

  • Nam, Gyeong-Hwa;Jang, Myoung-Uoon;Kim, Min-Jeong;Lee, Jung-Min;Lee, Min-Jae;Kim, Tae-Jip
    • Korean Journal of Microbiology
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    • v.52 no.4
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    • pp.463-470
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    • 2016
  • The enzymatic degradation of xylans is the most versatile way to obtain the high value-added functional compounds or the fermentable sugars for renewable energy. The endo-${\beta}$-xylanases are the major enzymes which hydrolyze the internal ${\beta}$-1,4-linkages of xylan backbones to produce the mixtures of xylooligosaccharides including xylobiose and xylotriose. Among them, glucuronoxylanase GH30 can exclusively hydrolyze the internal ${\beta}$-1,4-linkages of xylans decorated with methylglucuronic acid branches. In the present study, two xylanolytic enzyme (PaXN_10 and PaGuXN_30) genes were cloned from Paenibacillus amylolyticus KCTC 3005, and expressed in Escherichia coli, respectively. PaXN_10 (38.7 kDa) belongs to the endo-${\beta}$-xylanases GH10 family, while PaGuXN_30 (58.5 kDa) is a member of glucuronoxylanase GH30. They share the same optimal reaction conditions at $50^{\circ}C$ and pH 7.0. Enzymatic characterization proposed that P. amylolyticus can utilize the hardwood glucuronoarabinoxylans via the cooperative actions of xylanases GH10 and GH30. The extracellular PaGuXN_30 is secreted into the medium and hydrolyzes glucuronoarabinoxylans to release a series of aldouronic acid mixtures with a methylglucuronic acid branch. The resultant products being transported into the microbial cell are successively degraded into the smaller xylooligosaccharides by the intracellular PaXN_10, which will be utilized for the cellular metabolism.

The Relationship between Heme Oxygenase-1 Expression and Response to Cisplatin Containing Chemotherapy in Advanced Non-Small Cell Lung Cancer (진행성 비소세포폐암에서 Heme oxygenase-1 발현과 Cisplatin을 포함하는 항암화학요법의 치료반응과의 연관성)

  • Yang, Doo Kyung;Roh, Mee Sook;Lee, Kyung Eun;Kim, Ki Nam;Lee, Ki Nam;Choi, Pil Jo;Bang, Jung Hee;Kim, Bo Kyung;Seo, Hyo Rim;Kim, Min Ji;Kim, Seul Ki;Lee, Soo-Keol;Son, Choon Hee
    • Tuberculosis and Respiratory Diseases
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    • v.60 no.3
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    • pp.314-320
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    • 2006
  • Background : The overall response (20-30%) to chemotherapy in non-small cell lung cancer (NSCLC) is quite poor. Heme oxygenase-1 (HO-1) is the rate-limiting enzyme in heme degradation. There is increasing evidence suggesting that the induction of HO-1 might have an important protective effect against oxidative stress including cisplatin containing chemotherapy. This study retrospectively investigated the relationship between HO-1 expression and the response to chemotherapy containing cisplatinin advanced NSCLC patients. Material and Methods : The medical records including the responses to chemotherapy of fifty nine cases were evaluated retrospectively, and the tissue samples of these patients were immunohistochemically stained for HO-1. Results : Forty three of the fifty nine patients(72.8%) showed positive staining for HO-1 in their cancer tissues. There was no significant difference according to the cell type, stage and tumor size. In addition, there was no correlation between HO-1 expression and the responses to chemotherapy. Conclusion : HO-1 expression in tumor tissue dose not predict the response to cisplatin containing chemotherapy in advanced NSCLC. Further prospective studies with a larger number of patients will be needed to confirm these results.

The Role of Heme Oxygenase-1 in Lung Cancer Cells (폐암세포주에서 Heme Oxygenase-1의 역할)

  • Jung, Jong-Hoon;Kim, Hak-Ryul;Kim, Eun-Jung;Hwang, Ki-Eun;Kim, So-Young;Park, Jung-Hyun;Kim, Hwi-Jung;Yang, Sei-Hoon;Jeong, Eun-Taek
    • Tuberculosis and Respiratory Diseases
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    • v.60 no.3
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    • pp.304-313
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    • 2006
  • Background : Heme oxygenase-1 (HO-1) is an inducible enzyme that catalyzes the oxidative degradation of heme to form biliverdin, carbon monoxide (CO), and free iron. The current evidence has indicated a critical role of HO-1 in cytoprotection and also in other, more diverse biological functions. It is known that the high expression of HO-1 occurs in various tumors, and that HO-1 has an important role in rapid tumor growth because of its antioxidative and antiapoptotic effects. Therefore, the role of HO-1 was analyzed in human lung cancer cell lines, and especially in the A549 cell line. Material and Methods : Human lung cancer cell lines, i.e., A549, NCI-H23, NCI-H157 and NCI-H460, were used for this study. The expression of HO-1 in the untreated state was defined by Western blotting. ZnPP, which is the specific HO inhibitor we used, and the viability of cells were tested for by conducting MTT assaysy. The HO enzymatic activity, as determined via the bilirubin level, was also indirectly measured. Moreover, the generation of intracellular hydrogen peroxide (H2O2) was monitored fluorimetrically with using a scopoletin-horse radish peroxidase (HRP) assay and 2',7'-dichlorofluorescein diacetate (DCFH-DA). We have also transfected small HO-1 interfering RNA (siRNA) into A549 cells, and the apoptotic effects were evaluated by flow cytometric analysis and Western blotting. Results : The A549 cells had a greater expression of HO-1 than the other cell lines, whereas ZnPP significantly decreased the viability of the A549 cells more than the viability of the other lung cancer cells in a dose-dependant fashion. Consistent with the viability, the HO enzymatic activity also was decreased. Moreover, intracellular H2O2 generation via ZnPP was induced in a dose-dependent manner. Apoptotic events were, then induced in the HO-1 siRNA transfected A549 cells. Conclusion : HO-1 provides new important insights into the possible molecular mechanism of the antitumor therapy in lung cancer.

Field Tests for Assessing the Bioremediation Feasibility of a Trichloroethylene-Contaminated Aquifer (관측정 자연표류 실험을 통한 트리클로로에틸렌(Trichloroethylene) 오염 지하수의 생물학적 복원 타당성 연구)

  • Kim Young;Kim Jin-Wook;Ha Chul-Yoon;Kim Nam-Hee;Hong Kwang-Pyo;Kwon Soo-Yul;Ahn Young-Ho;Ha Joon-Su;Park Hoo-Won
    • Journal of Soil and Groundwater Environment
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    • v.10 no.3
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    • pp.38-45
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    • 2005
  • The feasibility of stimulating in situ aerobic cometabolic activity of indigenous microorganisms was investigated in a trichloroethylene (TCE)-contaminated aquifer. A series of single-well natural drift tests (SWNDTs) was conducted by injecting site groundwater amended with a bromide tracer and combinations of toluene, oxygen, nitrate, ethylene and TCE into an existing monitoring well and by sampling the same well over time. Three field tests, Push-pull Transport Test, Drift Biostimulation Test, and Drift Surrogate Activity Test, were performed in sequence. Initial rate of toluene degradation was much faster than the rate of bromide dilution resulting from natural groundwater drift, indicating stimulation of indigenous toluene-oxidizing microorganisms. Transformation of ethylene, a surrogate probing overall activity of TCE transformation, was also observed, and its transformation results in the production of ethylene oxide, suggesting that some tolueneoxidizing microorganisms stimulated may express a orthomonooxygenase enzyme. Also in situ transformation of TCE was confirmed by greater retardation of TCE than bromide after the stimulation of toluene-oxidizing microorganisms. These results indicate that, in this environment, toluene and oxygen additions stimulated the growth and aerobic cometabolic activity of indigenous microorganisms expressing orthomonooxygenase enzymes. The simple, low-cost field test method presented in this study provides an effective method for conducting rapid field assessments and pilot testing of aerobic cometabolism, which has previously hindered application of this technology to groundwater remediation.

Inhibitory Effects of Schisandrae Fructus Ethanol Extract on the Production of Matrix Metalloproteinases in in vitro and in vivo Osteoarthritis Models (In vitro 및 in vivo 퇴행성관절염 모델에서 오미자 에탄올 추출물에 의한 matrix metalloproteinases의 생성 억제)

  • Jeong, Jin-Woo;Lee, Hye Hyeon;Kim, Hong Jae;Lee, Ki Won;Kim, Ki Young;Kim, Sung Goo;Hong, Su-Hyun;Kim, Bum Hoi;Park, Cheol;Choi, Yung Hyun
    • Journal of Life Science
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    • v.27 no.10
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    • pp.1207-1214
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    • 2017
  • Schisandrae Fructus (SF), the fruit of Schisandra chinensis (Turcz.) Baill., is widely used in traditional medicine for the treatment of a number of chronic diseases. SF extracts have been recently reported to attenuate the inflammatory responses in SW1353 human chondrocyte cells in in vitro and monosodium iodoacetate (MIA)-induced cartilage degradation in in vivo osteoarthritis (OA) models. However, their protective and therapeutic potentials against OA in primary culture chondrocytes and animal models remain unclear. Therefore, we investigated the effects of the ethanol extract of SF on the activity of matrix metalloproteinases (MMPs), biomarkers for diagnosis of OA, on interleukin $(IL)-1{\beta}-induced$ primary cultured rat cartilage chondrocytes and MIA-induced osteoarthritis in a rat model. Our data indicated that SF treatment significantly reduced the mRNA expression and enzyme activity of MMP-1, -3 and -13 in $IL-1{\beta}-induced$ primary cultured rat cartilage chondrocytes. The chondro-protective effects of SF were then analyzed in a rat OA model using a single intra-articular injection of MIA in the right knee joint. According to our results, the elevated levels of MMP-1 and -3 were markedly ameliorated by SF administration. Collectively, these findings indicate that SF could be a candidate for the treatment of OA.

The effect of environmental condition to the mycelial browning of Lentinula edodes (Berkeley) Sing. during sawdust bag cultivation (환경조건이 표고톱밥배지의 갈변에 미치는 영향)

  • Kim, Young-Ho;Jhune, Chang-Sung;Park, Soo-Chul;You, Chang-Hyun;Sung, Jae-Mo;Kong, Won-Sik
    • Journal of Mushroom
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    • v.7 no.3
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    • pp.115-121
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    • 2009
  • Recently sawdust cultivation of Shiitake mushroom (Lentinula edodes ) is getting increased because log cultivation is getting difficult to get oak logs. It is important to make mycelia browning on the substrate surface in sawdust cultivation. This browned surface plays an important role like as artificial bark of the oak log, which protects the other pests and suppresses water evaporation in the substrate. The period for mycelia browning is so long that the sawdust cultivation of Shiitake mushroom can not spread well into the mushroom farms. In this article we would like to discuss about the effect of environmental condition to the mycelial browning during sawdust bag cultivation for the To reduce the period required for browning of substrates, sawdust substrates was illuminated light with difference intensity. One hundred Lux light illumination was needed for producing normal yield of fruit body but fruit body yield was low and abnormally shaped fruit body was produced when cultured under the dark condition of incubation. Illumination over 200lux is necessary for the successful browning of substrates during incubation. Optimum incubation temperature for browning of substrates and fruiting was $25^{\circ}C$. The treatment of cotton plug with different size to identify the effect of aeration on the browning of substrates and fruiting showed rapid mycelial growth and reduced the periods for browning as the size of cotton plug was bigger. However, yield of fruit body was the highest at 16mm diameter cotton plug as compared to 20mm of that. $CO_2$ content in vessel of substrates was low as the size of cotton plug was bigger during incubation. $CO_2$ content during incubation of substrate was highest in periods between 8 week and 14 week after inoculation of shiitake when substrate was changed color into brown. $C_2H_4$ content in vessel with substrates was highest at 8mm diameter cotton plug and it was increased by order of 12, 16, 20, 0, 4 mm diameter cotton plug during substrate incubation. Sawdust substrate was soaked in cold water for different time to identify soaking effect of sawdust substrate on fruit body yield and activities of enzymes in these substrates were investigated. The fruit body yield was increased up to 40% by soaking substrates in comparison with unsoaked substrates. The soaked substrates showed 165, 175g/1,000ml at treatment of 4 and 15 hours, respectively. Cellulose activities in soaked substrates were not changed with soaking time, but activities of laccase, lignin degradation enzyme, were drastically increased up to 4 times in comparison with unsoaked substrates.

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Promoter -202 A/C Polymorphism of Insulin-like Growth Factor Binding Protein-3 Gene and Non-small Cell Lung Cancer Risk (인슐린양 성장 인자 결합 단백-3 유전자 -202 좌위의 다형성에 따른 비소세포폐암의 위험도)

  • Moon, Jin Wook;Chang, Yoon Soo;Han, Chang Hoon;Kang, Shin Myung;Park, Moo Suk;Byun, Min Kwang;Chung, Wou Young;Park, Jae Jun;Yoo, Kyeong Nam;Shin, Ju Hye;Kim, Young Sam;Chang, Joon;Kim, Sung Kyu;Kim, Hee Jung;Kim, Se Kyu
    • Tuberculosis and Respiratory Diseases
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    • v.58 no.4
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    • pp.359-366
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    • 2005
  • Background : IGFBP-3 inhibits the mitogenic and anti-apoptotic activity of IGF by blocking the binding of IGF to its receptor. However, under certain circumstances, IGFBP-3 can enhance the activity of IGF by protecting IGF from its degradation. More than half of the interindividual variations in IGFBP-3 levels are known to be genetically determined by the polymorphism at -202 locus of IGFBP-3 gene. Method : We attempted to ascertain whether A-202C polymorphic variation of IGFBP-3 gene constitutes a risk factor for non-small cell lung cancer (NSCLC), using PCR-restriction fragment length polymorphism (RFLP). Our study included 104 NSCLC patients and 104 age-, gender-, and smoking status-matched control subjects. Result : In the 104 NSCLC subjects, the genotypic frequencies at the -202 site were as follows: AA = 67 (64.4%), AC = 35 (33.7%), and CC = 2 (1.9%). We did detect significant differences in the genotypic distribution between the NSCLC and the control subjects (p<0.05), and the NSCLC risk correlated significantly with AA genotype at the -202 locus (AA>AC>CC). Using CC genotype as a reference, the odds ratio (OR) for the subjects with AC genotype was 2.60 (95% CI: 0.89 - 8.60), and the OR associated with AA genotype was 5.89 (95% CI: 1.92 - 21.16). Conclusion : These results indicate that the dysregulation of IGF axis should now be considered as another important risk factor for NSCLC, and a potential target for novel antineoplastic therapies and/or preventative strategies in high-risk groups.

The change of grain quality and starch assimilation of rice under future climate conditions according to RCP 8.5 scenario (RCP 8.5 시나리오에 따른 미래 기후조건에서 벼의 품질 및 전분 동화 특성 변화)

  • Sang, Wan-Gyu;Cho, Hyeoun-Suk;Kim, Jun-Hwan;Shin, Pyong;Baek, Jae-Kyeong;Lee, Yun-Ho;Cho, Jeong-Il;Seo, Myung-Chul
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.20 no.4
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    • pp.296-304
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    • 2018
  • The objective of this study was to analyze the impact of climate change on rice yield and quality. Experiments were conducted using SPAR(Soil-Plant-Atmosphere-Research) chambers, which was designed to create virtual future climate conditions, in the National Institute of Crop Science, Jeonju, Korea, in 2016. In the future climate conditions($+2.8^{\circ}C$ temp, 580 ppm $CO_2$) of year 2051~2060 according to RCP 8.5 scenario, elevated temperature and $CO_2$ accelerated the heading date by about five days than the present climate conditions, resulted in a high temperature environment during grain filling stage. Rice yield decreased sharply in the future climate conditions due to the high temperature induced poor ripening. And the spikelet numbers, ripening ratio, and 1000-grain weight of brown rice were significantly decreased compared to control. The rice grain quality was also decreased sharply, especially due to the increased immature grains. In the future climate conditions, expression of starch biosynthesis-related genes such as granule-bound starch synthase(GBSSI, GBSSII, SSIIa, SSIIb, SSIIIa), starch branching enzyme(BEIIb) and ADP-glucose pyrophosphorylase(AGPS1, AGPS2, AGPL2) were repressed in developing seeds, whereas starch degradation related genes such as ${\alpha}-amylase$(Amy1C, Amy3D, Amy3E) were induced. These results suggest that the reduction in yield and quality of rice in the future climate conditions is likely caused mainly by the poor grain filling by high temperature. Therefore, it is suggested to develop tolerant cultivars to high temperature during grain filling period and a new cropping system in order to ensure a high quality of rice in the future climate conditions.