• Title/Summary/Keyword: glucanase

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Molecular Cloning and Sequence Analysis of Coelomic Cytolytic Factor-like Gene from the Midgut of the Earthworm, Eisenia Andrei (줄지렁이 중장에서 분리한 Coelomic cytolytic factor-유사 유전자의 클로닝 및 염기서열 분석에 관한 연구)

  • Baek, Nam Sook;Lee, Myung-Sik;Park, Sang-Kil;Kim, Dae-hwan;Tak, Eun-Sik;Ahn, Chi-Hyun;Sun, Zhenjun;Park, Soon Cheol
    • Journal of the Korea Organic Resources Recycling Association
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    • v.16 no.4
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    • pp.64-73
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    • 2008
  • The cDNA of CCF (coelomic cytolytic factor)-like gene (EC 3.2.1.16), a kind of glycosyl hydorlase, was isolated and cloned from the midgut of the earthworm Eisenia anderi. The size of nucleotide sequence appeared to be 1,152 bp and its predicted coding region was composed of 384 amino acid residues including the initiation methionine. The 17 residues at N-terminal end in the deduced amino acid sequence were regarded to be a signal peptide. Based on the amino acid sequence analysis, it appeared that this CCF-like protein could belong to glycosyl hydrolase family 16 (GHF16) and showed a high sequence homology of about 79~99% with CCF and CCF-like proteins from other earthworm species. The CCFs and CCF-like proteins from various earthworm species exhibited a 100% homology in the polysacchride-binding motif and glucanase motif. It has been reported that the CCFs isolated from E. fedita appeared to show a broader pattern recognition specificity than those from other earthworm species because this species resides in decaying organic matter showing very high microbial activity, implying that CCF-like protein isolated in this study from E. andrei might exhibit a broad substrate specificity that is a useful characteristic for industrial application. A phylogenetic analysis using the deduced amino acid sequences of CCF-related proteins through the BLASTX revealed that GHF16 families could be divided into three groups of metazoa, viriplantae and eubacteria subfamily. Subsequently the CCF-related proteins of metazoa subfamily could clearly be subgroup into lophotrochozoan and edysozoan type including a deuterostome origin. Further understanding of the biological properties of E. andrei CCF-like protein should be addressed to regulate the ${\beta}$-D-glucan hydrolysis and production for the industrial uses.

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Effect of Rice Flour Prepared with Enzyme Treatment on Quality Characteristics of Rice Cookies (효소처리 쌀가루를 이용한 쌀 쿠키의 품질 특성)

  • Kim, Mi-Seon;Park, Jong Dae;Lee, Hyun Yu;Kum, Jun Seok
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.9
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    • pp.1439-1445
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    • 2013
  • The aim of this study was to investigate the quality characteristics of rice cookies specifically prepared with rice flour following enzyme treatment. In this study, three types of specific enzyme treatments, that is, ${\alpha}$-amylase, ${\beta}$-amylase, and cellulose+${\beta}$-glucanase were applied on the effect of rice flour with respect to desired quality characteristics of the rice cookies. Based on our study results, we have found that the density of dough in the rice cookies was not significantly different between that of the non-enzyme treated (control) and the enzyme treated group. However, we found that the spread factor of rice cookies, prepared with rice flour treated with ${\beta}$-amylase, was higher than that of the control. It was also found that the moisture content of rice cookies (with added enzyme-treated rice flour) was 3.20 to 3.90%; however, this range is much lower than that observed in the control. Further, we observed that Hunter color's L-values were significantly higher for the control than those of the enzyme-treated cookies. We also found that hardness of enzyme-treated cookies was comparatively better than that of the control. In addition, the sensory acceptability scores of the enzyme-treated cookies were found to be significantly higher than the control in decisive parameters such as aroma, appearance, flavor, taste, texture, and overall acceptability. Based on our findings, we suggest that ${\alpha}$-amylase treated rice flour is an effective ingredient for improving the overall quality of rice cookies.

Effects of Exogenous Enzymes on Ruminal Fermentation and Degradability of Alfalfa Hay and Rice Straw

  • Yang, H.E.;Son, Y.S.;Beauchemin, K.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.1
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    • pp.56-64
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    • 2011
  • This study was conducted to evaluate the use of exogenous enzymes as a potential means of improving the ruminal digestion (i.e., degradability) of alfalfa hay and rice straw. Twenty six enzyme-additives were examined in terms of protein concentration and enzymic activities on model substrates. The exogenous enzymes contained ranges of endoglucanase, xylanase, ${\beta}$-glucanase, ${\alpha}$-amylase, and protease activities. Six of the enzyme additives were chosen for further investigation. The enzyme additives and a control without enzyme were applied to mature quality alfalfa hay substrate and subsequently incubated in rumen batch cultures. Five of the enzyme additives (CE2, CE13, CE14, CE19, and CE24) increased total gas production (GP) at 48 h of incubation compared to the control (p<0.05). The two additives (CE14 and CE24) having the greatest positive effects on alfalfa hay dry matter, neutral detergent fibre (NDF) and acid detergent fibre (ADF) degradability were further characterized for their ability to enhance degradation of low quality forages. The treatments CE14, CE24, a 50:50 combination of CE14 and CE24 (CE14+24), and control (no enzyme) were applied to mature alfalfa hay and rice straw. For alfalfa hay, application of the two enzyme additives, alone and in combination, increased GP compared to the control at 48 h fermentation (p<0.05), whereas only CE14 and CE14+24 treatments improved GP from rice straw (p<0.05). Rumen fluid volatile fatty acid concentrations throughout the incubation of rice straw were analyzed. Acetate concentration was slightly lower (p<0.05) for CE14${\times}$CE24 compared to the control, although individually, CE14 and CE24 acetate concentrations were not different from the control. Increases (p<0.05) in alfalfa hay NDF degradability measured at 12 and 48 h of incubation occurred only for CE14 (at 12 h) and for CE14+24 (at 12 and 48 h). Similarly, ADF degradability increased (p<0.05) with CE14 and CE14+24. As for rice straw, increased DM degradability was observed at 12 and 48 h of incubation for all enzyme treatments with an exception for CE14 at 12 h. The degradability of NDF was improved by all the enzyme treatments at either incubation time, while ADF degradability was only enhanced at 48 h. Overall, the enzymes led to enhanced digestion of mature alfalfa and there was evidence of improved digestibility of rice straw, an even lower quality forage.

Effects of dietary enzyme cocktail on diarrhea and immune responses of weaned pigs

  • Kang, Joowon;Cho, Jeeyeon;Jang, Kibeom;Kim, Junsu;Kim, Sheena;Mun, Daye;Kim, Byeonghyeon;Kim, Younghwa;Park, Juncheol;Choe, Jeehwan;Song, Minho
    • Korean Journal of Agricultural Science
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    • v.44 no.4
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    • pp.525-530
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    • 2017
  • Weaning is the most stressful event for nursery pigs because they are moved from familiar to unfamiliar environments. In addition, weaned pigs have immature digestive and immune systems. This situation makes weaned pigs susceptible to diseases and makes the absorption of nutrients from diets difficult. A feed approach, such as dietary enzyme supplementation, can be considered a solution. This study investigated the effects of dietary enzyme cocktail on diarrhea and immune responses of weaned pigs. A total 36 weaned pigs ($5.92{\pm}0.48kg\;BW$; 28 d old) were randomly allotted to 2 dietary treatments (3 pigs/pen, 6 replicates/treatment) in a randomized complete block design. The dietary treatments were a typical diet based on corn and soybean meal (CON) and CON with 0.05% enzyme cocktail (Cocktail; combination of xylanase, ${\alpha}-amylase$, protease, ${\beta}-glucanase$, and pectinase). Pigs were fed their respective diets for 6 wk. Incidence of diarrhea, packed cell volume (PCV), white blood cells (WBC) count, and immunoglobulin content were measured. A significantly lower incidence of diarrhea (p < 0.05) was observed in the Cocktail group as compared with the CON group. The Cocktail group also showed a decreased PCV (p < 0.1) on d 3 after weaning than the CON group. However, no differences were observed for number of WBC and contents of immunoglobulin G, M, and A between the Cocktail and CON groups. Consequently, inclusion of an enzyme cocktail in diets for weaned pigs had a positive influence on gut health by reducing the incidence of diarrhea in the present study.

Effects of dietary enzyme cocktail on growth performance, intestinal morphology, and nutrient digestibility of weaned pigs

  • Kim, Yunkang;Baek, Jangryeol;Jang, Kibeom;Kim, Junsu;Kim, Sheena;Mun, Daye;Kim, Byeonghyeon;Kim, Younghwa;Park, Juncheol;Choe, Jeehwan;Song, Minho
    • Korean Journal of Agricultural Science
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    • v.44 no.4
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    • pp.513-518
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    • 2017
  • Soybean, one of most widely used swine feed component in the world, contains non-starch polysaccharides (NSP). The digestive system of weaned pigs is not yet fully developed, and thus weaned pigs cannot easily digest diets based on corn and soybean meal. Dietary exogenous enzymes supplementation has been intensively investigated to assist digestion of anti-nutritional factors, such as NSP. This experiment was conducted to investigate the effects of dietary enzyme cocktail on growth performance, intestinal morphology, and nutrient digestibility of weaned pigs. A total 36 weaned pigs ($5.92{\pm}0.48kg\;BW$; 28 d old) were randomly allotted to 2 dietary treatments (3 pigs/pen, 6 replicates/treatment) in a randomized complete block design. The dietary treatments were a typical diet based on corn and soybean meal (CON) and CON with 0.05% enzyme cocktail (Cocktail; mixture of xylanase, ${\alpha}-amylase$, protease, ${\beta}-glucanase$, and pectinase). Pigs were fed their respective diets for 6 wk. Growth performance, morphology of ileum, apparent ileal digestibility (AID) and apparent total tract digestibility (ATTD) of dry matter, crude protein, and energy of weaned pigs were measured. No significant differences (p > 0.05) were observed for growth performance for the duration of the experimental period, and morphology of ileum, and nutrient digestibility between CON and Cocktail treatment groups. Therefore, the results from the current study indicated that enzyme cocktail supplementation in diets had no influence on growth performance, intestinal morphology, and nutrient digestibility of weaned pigs.

Proteomic analysis of Korean ginseng(Panax ginseng C. A. Meyer) following exposure to salt stress

  • Kim, Sun-Tae;Bae, Dong-Won;Lee, Kyung-Hee;Hwang, Jung-Eun;Bang, Kyong-Hwan;Kim, Young-Chang;Kim, Ok-Tae;Yoo, Nam-Hee;Kang, Kyu-Young;Hyun, Dong-Yun;Lim, Chae-Oh
    • Journal of Plant Biotechnology
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    • v.35 no.3
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    • pp.185-193
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    • 2008
  • We evaluated the response to salt stress of two different ginseng lines, STG3134 and STG3159, which are sensitive and tolerant, respectively, to salt treatment. Plants were exposed to a 5 dS/m salt solution, and chlorophyll fluorescence was measured. STG3134 ginseng was more sensitive than STG3159 to salt stress. To characterize the cellular response to salt stress in the two different lines, changes in protein expression were investigated using a proteomic approach. Total protein was extracted from detached salt-treated leaves of STG3134 and STG3159 ginseng, and then separated by two-dimensional polyacrylamide gel electrophoresis(2-DE). Approximately 468 protein spots were detected by 2-DE and Coommassie brilliant blue staining. Twenty-two proteins were found to be reproducibly up- or down-regulated in response to salt stress. Among these proteins, twelve were identified using MALDI-TOF MS and ESI-Q-TOF and classified into several functional groups: photosynthesis-related proteins(oxygen-evolving enhancer proteins 1 and 2, rubisco and rubisco activase), detoxification proteins(polyphenol oxidase) and defense proteins($\beta$-1,3-glucanase, ribonuclease-like storage protein, and isoflavone reductase-like protein). The protein levels of ribonuclease-like storage protein, which was highly induced in STG3159 ginseng as compared to STG3134, correlated tightly with mRNA transcript levels, as assessed by reverse-transcription(RT)-PCR. Our results indicate that salinity induces changes in the expression levels of specific proteins in the leaves of ginseng plants. These changes may, in turn, playa role in plant adaptation to saline conditions.

Quantitative Changes of PR Proteins and Antioxidative Enzymes in Response to Glomus intraradices and Phytophthora capsici in Pepper (Capsicum annuum L.) Plants

  • Zheng, Hu-Zhe;Kim, Yong-Woong;Lee, Hyun-Jin;Park, Ro-Dong;Jung, Woo-Jin;Kim, Young-Cheol;Lee, Sang-Hyun;Kim, Tae-Hwan;Kim, Kil-Yong
    • Journal of Microbiology and Biotechnology
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    • v.14 no.3
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    • pp.553-562
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    • 2004
  • To investigate protective activity in pepper plants, which were pre-inoculated with arbuscular mycorrhizal (AM) fungi Glomus intra radices (Gi), against pathogenic strain Phytophthora capsici (Pc), pathogenesis-related (PR) proteins and antioxidant enzymes were examined. The growth of root and shoot was the highest in peppers inoculated with G. intraradices, compared with non-inoculated control plants and those challenged by the pathogen with and without mycorrhizae after nine days of infection. Mycorrhizal colonization rate was reduced by about 10% in pathogen-challenged plants, but disease pressure was reduced. The activities of PR proteins, $\beta$-1- 3-glucanase and chitinase, were increased in Pc-treated plants compared to Gi+Pc-treated plants in leaves, but those in roots were suppressed. Superoxide dismutase activity and $H_2O_2${/TEX> content in Gi+Pc and Pc-treated plants were gradually increased in leaves. However, those in roots continuously increased up to 5 days, and then decreased dramatically. Peroxidase activity in leaves and roots increased after P. capsici infection both in plants inoculated with or without G. intraradices. These results suggest that AM fungi, G. intra radices, potentially act as one of the protective agents against plant pathogens. Changes of PR proteins and antioxidative enzymes in mycorrhizae-inoculated pepper appear to be regulated differently in leaves and roots by pathogen infection.

Effects of dietary carbohydrases on productive performance and immune responses of lactating sows and their piglets

  • Lee, Jeong Jae;Choi, Seong Ho;Cho, Jin Ho;Choe, Jeehwan;Kang, Joowon;Kim, Soyun;Park, Sangwoo;Kyoung, Hyunjin;Seo, Dongoh;Cho, Jee-Yeon;Park, Il-Hun;Oh, Sangnam;Kim, Hyeun Bum;Song, Minho
    • Journal of Animal Science and Technology
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    • v.61 no.6
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    • pp.359-365
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    • 2019
  • This study was conducted to evaluate effects of dietary multi-carbohydrases (MCS) in a lactating sow diet on productive performance and immune responses of sows and their piglets. A total of 12 sows (218.37 ± 5.5 kg BW; 2 parity) were randomly assigned to 2 dietary treatments: a diet based on corn-soybean meal (CON) and CON with 0.01% MCS. The MCS contained xylanase (2,700 units/g), β-glucanase (700 units/g), and cellulase (800 units/g). Sows were fed the dietary treatments for 28 days (weaning) after farrowing. Blood samples were collected from sows on d 0, 3, and 7 after farrowing and randomly selected 2 nursing piglets in each sow on d 3, 7, and 14 after birth. Measurements were productive performance of sows, frequency of diarrhea of piglets, and immune responses of sows and their piglets. Sows fed MCS had lower (p < 0.05) their body weight change than those fed CON. Piglets from sows fed MCS had higher (p < 0.05) average weight gain and body weight at weaning day and lower (p < 0.10) frequency of diarrhea than those from sows fed CON. Sows fed MCS had lower number of white blood cells (WBC) on d 3 (p < 0.05) and TGF-β1 on d 7 (p < 0.10) during lactation than those fed CON. Similarly, piglets from sows fed MCS had also lower (p < 0.05) number of WBC on d 3 and d 7 and TGF-β1 and C-reactive protein on d 7 during lactation than those from sows fed CON. In addition, piglets from sows fed MCS had higher (p < 0.10) immunoglobulin G and M on d 7 during lactation those from sows fed CON. In conclusion, addition of dietary MCS in the lactating sow diet based on corn and soybean meal improved productive performance of sows and their litters and modulated their immune responses.

Effect of Exogenous Fibrolytic Enzyme Application on the Microbial Attachment and Digestion of Barley Straw In vitro

  • Wang, Y.;Ramirez-Bribiesca, J.E.;Yanke, L.J.;Tsang, A.;McAllister, T.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.1
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    • pp.66-74
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    • 2012
  • The effects of exogenous fibrolytic enzymes (EFE; a mixture of two preparations from Trichoderma spp., with predominant xylanase and ${\beta}$-glucanase activities, respectively) on colonization and digestion of ground barley straw and alfalfa hay by Fibrobacter succinogenes S85 and Ruminococcus flavefaciens FD1 were studied in vitro. The two levels (28 and 280 ${\mu}g$/ml) of EFE tested and both bacteria were effective at digesting NDF of hay and straw. With both substrates, more NDF hydrolysis (p<0.01) was achieved with EFE alone at 280 than at 28 ${\mu}g$/ml. A synergistic effect (p<0.01) of F. succinogenes S85 and EFE on straw digestion was observed at 28 but not 280 ${\mu}g$/ml of EFE. Strain R. flavefaciens FD1 digested more (p<0.01) hay and straw with higher EFE than with lower or no EFE, but the effect was additive rather than synergistic. Included in the incubation medium, EFE showed potential to improve fibre digestion by cellulolytic ruminal bacteria. In a second batch culture experiment using mixed rumen microbes, DM disappearance (DMD), gas production and incorporation of $^{15}N$ into particle-associated microbial N ($^{15}N$-PAMN) were higher (p<0.001) with ammoniated (5% w/w; AS) than with native (S) ground barley straw. Application of EFE to the straws increased (p<0.001) DMD and gas production at 4 and 12 h, but not at 48 h of the incubation. EFE applied onto S increased (p<0.01) $^{15}N$-PAMN at 4 h only, but EFE on AS increased (p<0.001) $^{15}N$-PAMN at all time points. Prehydrolysis increased (p<0.01) DMD from both S and AS at 4 and 12 h, but reduced (p<0.01) $^{15}N$-PAMN in the early stage (4 h) of the incubation, as compared to non-prehydrolyzed samples. Application of EFE to barley straw increased rumen bacterial colonization of the substrate, but excessive hydrolytic action of EFE prior to incubation decreased it.

Acetone, Butanol, Ethanol Production from Undaria pinnatifida Using Clostridium sp. (Clostridium 종을 이용한 미역으로부터 아세톤, 부탄올, 에탄올 (ABE) 생산)

  • Kwon, Jeong Eun;Gwak, Seung Hee;Kim, Jin A;Ryu, Ji A;Park, Sang Eon;Baek, Yoon Seo;Heo, A Jeong;Kim, Sung-Koo
    • Microbiology and Biotechnology Letters
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    • v.45 no.3
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    • pp.236-242
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    • 2017
  • The conversion of marine biomass to renewable energy has been considered an alternative to fossil fuels. Butanol, in particular, can be used directly as a fuel. In this experiment, the brown alga Undaria pinnatifida was selected as a biomass for biobutanol production. Hyper thermal (HT) acid hydrolysis was used as an acid hydrolysis method to produce monosaccharides. The optimal pretreatment conditions for U. pinnatifida were determined as slurry with 10% (w/v) U. pinnatifida content and 270 mM $H_2SO_4$, and heating at $160^{\circ}C$ for 7.5 min. Enzymatic saccharification was carried out with Celluclast 1.5 L, Viscozyme L, and Ultraflo Max. The optimal saccharification condition was 12 U/ml Viscozyme L. Fermentations were carried out for the production of acetone, butanol, and ethanol by Clostridium acetobutylicum KCTC 1724, Clostridium beijerinckii KCTC 1785, and Clostridium tyrobutyricum KCTC 5387. The fermentations were carried out using a pH-control. The optimal ABE fermentation condition determined using C. acetobutylicum KCTC 1724 adapted to 160 g/l mannitol. An ABE concentration of 9.05 g/l (0.99 g/l acetone, 5.62 g/l butanol, 2.44 g/l ethanol) was obtained by the consumption of 24.14 g/l monosaccharide with $Y_{ABE}$ of 0.37 in pH 5.0.