• Title/Summary/Keyword: $In$ $vitro$ degradation

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Enzymatic in vitro glycosylation using peptide-N-glycosidase F

  • Lee, Ji-Yeon;Park, Tae-Hyeon
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.721-724
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    • 2000
  • The possibility of the enzymatic in vitro glycosylation using peptide-N-glycosidase F was examined. Oligosaccharide chains in the glycoproteins are important for the biological activity, solubility, immunogenecity, recognition, and prevention of degradation. After 4 h incubation of deglycosylated glycoprotein with excess glucose oligomer and ammonia in acetone at $50^{\circ}C$, upper shift of protein band was observed on SDS-PAGE. And the different deglycosylation characteristics of glucose oxidase and fetuin were investigated.

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Antioxidative Effects of Cichorium intybus Root Extract on LDL (Low Density Lipoprotein) Oxidation

  • Kim, Tae-Woong;Yang, Ki-Sook
    • Archives of Pharmacal Research
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    • v.24 no.5
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    • pp.431-436
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    • 2001
  • The water extract of Cichorium intybus (WECI) showed a remarkable antioxidative effect on LDL, and inhibitory effects on the production of thiobarbituric acid reactive substance and the Degradation of fatty acids in LDL. Vitamin 1 and unsaturated fatty acids in LDL were protected by adding WECI from the effects of metal catalyzed LDL oxidation. From the results obtained, we conclude that LDL oxidation is inhibited in vitro by the addition of WECI, and that LDL is protected by WECI from oxidative attack, as shown by agarose gel electrohporesis.

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Evaluation of Different Yeast Species for Improving In vitro Fermentation of Cereal Straws

  • Wang, Zuo;He, Zhixiong;Beauchemin, Karen A.;Tang, Shaoxun;Zhou, Chuanshe;Han, Xuefeng;Wang, Min;Kang, Jinhe;Odongo, Nicholas E.;Tan, Zhiliang
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.2
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    • pp.230-240
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    • 2016
  • Information on the effects of different yeast species on ruminal fermentation is limited. This experiment was conducted in a $3{\times}4$ factorial arrangement to explore and compare the effects of addition of three different live yeast species (Candida utilis 1314, Saccharomyces cerevisiae 1355, and Candida tropicalis 1254) at four doses (0, $0.25{\times}10^7$, $0.50{\times}10^7$, and $0.75{\times}10^7$ colony-forming unit [cfu]) on in vitro gas production kinetics, fiber degradation, methane production and ruminal fermentation characteristics of maize stover, and rice straw by mixed rumen microorganisms in dairy cows. The maximum gas production (Vf), dry matter disappearance (IVDMD), neutral detergent fiber disappearance (IVNDFD), and methane production in C. utilis group were less (p<0.01) than other two live yeast supplemented groups. The inclusion of S. cerevisiae reduced (p<0.01) the concentrations of ammonia nitrogen ($NH_3$-N), isobutyrate, and isovalerate compared to the other two yeast groups. C. tropicalis addition generally enhanced (p<0.05) IVDMD and IVNDFD. The $NH_3$-N concentration and $CH_4$ production were increased (p<0.05) by the addition of S. cerevisiae and C. tropicalis compared with the control. Supplementation of three yeast species decreased (p<0.05) or numerically decreased the ratio of acetate to propionate. The current results indicate that C. tropicalis is more preferred as yeast culture supplements, and its optimal dose should be $0.25{\times}10^7$ cfu/500 mg substrates in vitro.

Effects of harvest time and added molasses on nutritional content, ensiling characteristics and in vitro degradation of whole crop wheat

  • Xia, Chuanqi;Liang, Yixun;Bai, Sarula;He, Yang;Muhammad, Aziz Ur Rahman;Su, Huawei;Cao, Binghai
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.3
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    • pp.354-362
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    • 2018
  • Objective: Wheat is an alternative to corn silage for ruminant feeding in northern China. This study examined the effects of harvest time and added molasses on nutritional content, ensiling characteristics and in vitro degradation of whole crop wheat (WCW). Methods: Fresh WCW at the milk-ripe stage was harvested at 0700 h (i.e., in the morning [Mo]) and 1700 h (i.e., in the afternoon [Af]), and then immediately used to prepare silage and make hay. Commercial molasses was added to Af WCW at 0%, 2%, 4%, and 6% (fresh weight) proportions. The WCW treated with molasses was mixed thoroughly prior to ensiling. Results: Dry matter (DM), neutral detergent fiber, water soluble carbohydrate (WSC) content (p<0.01), accumulative gas production in 72 h ($GP_{72h}$, 77.46 mL/g vs 95.15 mL/g) and dry matter disappearance in vitro (69.15% vs 76.77%) were lower (p<0.05), while crude protein (CP) content was higher for WCW silage (WCWS) compared to WCW (p<0.01). The propionic acid and butyric acid concentrations in WCWS from Mo WCW were 1.47% and 0.26%, respectively. However, the propionic and butyric acid concentrations were negligible, while the ammonia nitrogen/total nitrogen ($NH_3-N/TN$, p<0.01) concentration was lower and the rate of gas production at 50% of the maximum (17.05 mL/h vs 13.94 mL/h, p<0.05) was higher for Af WCWS compared to Mo WCWS. The incubation fluid's $NH_3-N$ concentration was lower in WCWS and Af WCW compared to Mo WCW (p<0.05). The CP and WSC content increased with increasing molasses levels (p<0.05). Furthermore, the pH (p<0.01) and time when gas production was 50% of the maximum (2.78 h vs 3.05 h, p<0.05) were lower in silage treated with 4% molasses than silage without molasses. Conclusion: Harvesting wheat crops in the afternoon and adding molasses at 4% level to WCW optimally improved ensiling characteristics, leading to well-preserved silage.

Evaluation of Anastomotic Strength and in-vitro Degradability with Microvascular Anastomosis Coupler Based on Injection Molding Condition made by Biodegradable Polycaprolactone(PCL) (생체분해성 폴리카프로락톤(PCL) 미세혈관 문합커플러의 사출성형조건에 따른 문합강도 및 in-vitro 분해능 평가)

  • Ahn, Geun-Seon;Han, Gig-Bong;Oh, Seung-Hyun;Park, Jong-Woong;Kim, Cheol-Woong
    • Transactions of the KSME C: Technology and Education
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    • v.1 no.2
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    • pp.167-177
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    • 2013
  • The use of mechanical anastomosis coupler instead of sutures has been increasing in microvascular anastomosis surgery. However, non-biodegradable anastomosis coupler has problems such as not only inflammatory reaction but also remaining permanently in operation wound. Therefore, we fabricated biodegradable anastomosis coupler using injection molding process to overcome the limitation of non-biodegradable anastomosis coupler. In various injection molding process conditions, the shrinkage was calculated with different cylinder temperatures and injection molding pressures and anastomotic strength was measured. As a result, changes in shrinkage hole part larger than the pin part. In addition, the shrinkage in the cylinder at higher temperatures increase. However, the higher the injection pressure, shrinkage tends to decrease, respectively. In-vitro degradation behavior of PCL anastomotic coupler evaluated for 12 weeks, water uptake was increased and molecular weight was accompanied by a reduction in mass of the biological degradation and reduction of anastomotic strength was confirmed. However, decreased levels of anastomotic strength enough to exceed the requirements of preclinical surgery, PCL microvascular anastomosis coupler suitable candidate materials that could identify.

In vitro Fermentation of Rumen Microorganisms Cultured in Medium Supplemented with Bacterio-mineral Water (BMW) Produced from Bio-reacted Swine Manure

  • Kim, Chang-Hyun;Park, Joong Kook;Lee, Gi Yeong;Seo, In Joon
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.10
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    • pp.1435-1439
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    • 2005
  • Bacterio-mineral water (BMW) produced from manure has been known to exert a number of positive effects on animal production and odor control. An experiment was conducted to examine the effects of BMW produced from bio-reacted swine manure on in vitro gas production, cellulose degradation, microbial growth and fibrolytic enzyme activities of mixed rumen microorganisms. The five levels of 0, 0.001, 0.005, 0.01 and 1.0% BMW were supplemented into serum vials containing mixed rumen microorganisms. Incubations were carried out anaerobically at $39^{\circ}C$ without shaking for 0, 12, 24, 48, 72 and 96 h. There were no significant (p>0.05) differences among the treatments for the initial rate of gas production. At 72 h incubation, the gas production tended (p<0.1) to be increased by the 0.01 and 1.0% BMW treatments compared with control and the 0.001% BMW treatment. At the end of incubation (96 h), the sample supplemented with 0.01% BMW was higher (p<0.05) than control (0% BMW) in the gas production. The microbial growth rate was increased by all the BMW treatments, while 0.01% BMW was most effective in stimulating the growth rate. Although the addition of BMW on the filter paper DM degradation was not significantly influenced throughout the incubation period except the 48 h incubation, DM degradation tended to be increased by all BMW treatments compared with control. The addition of both 0.005 and 0.01% BMW highly increased (p<0.05) CMCase activity compared with control after 24 h and 48 h incubation, while at the 72 h incubation the 0.01% BMW addition only significantly increased (p<0.05). After 72 h incubation, the xylanase activity was significantly (p<0.05) increased with the addition of 1.0% BMW compared with the addition of 0.001 and 0.005% BMW, while at the other incubation times, the xylanase activity was not different among the treatments. In conclusion, the 0.01% BMW of supplementation level would be the suitable addition level to stimulate rumen fermentation increasing microbial growth and cellulose degradation.

Effects of Replacing Rice Straw with By-products of Medical Herbs on the in vitro Fermentation Characteristics (한약재 부산물의 대체 수준이 in vitro 발효특성에 미치는 영향)

  • Lee, Shin-Ja;Shin, Nyeon-Hak;An, Jung-Jun;Chu, Gyo-Moon;Moon, Yea-Hwang;Lee, Sung-Sill
    • Journal of agriculture & life science
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    • v.45 no.3
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    • pp.69-79
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    • 2011
  • This study was conducted to estimate effects of by-products of medical herbs replacing rice straw on in vitro fermentation characteristics. Each trial was composed of five treatments including medical herbs : rice straw (%) = 20 : 80 (T1), 40 : 60 (T2), 50 : 50 (T3), 100 : 0 (T4) and the control. Each treatment had eight fermentation times (3, 6, 9, 12, 24, 36, 48 and 72 hours) with three replications. The gas production and DM degradation were significantly (P<0.05) increased by supplementation, especially T4, during the whole fermentation periods. Methane production increased along with addition of by-products similar to the gas production and DM degradation. The pH values ranged from 5.39 to 6.80 and were significantly (P<0.05) decreased by supplementation of by-products of medical herbs. Microbial growth rates reached the peak at between 36 and 48h, thereafter tended to decrease. Although there were no significant differences in the enzyme activities, there was a tendency of increase in T4 treatment. From above results, the replacement levels, particularly 100% replacement of rice straw by by-products of medical herbs, resulted in improving the in vitro fermentation characteristics such as increasing gas production, microbial growth and DM degradation. Also it may help digestion by increasing enzyme activities.

Effects of Methylcellulose on Fibrolytic Bacterial Detachment and In vitro Degradation of Rice Straw

  • Kim, Min Ji;Sung, Ha Guyn;Upadhaya, Santi Devi;Ha, Jong K.;Lee, Sung Sill
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.10
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    • pp.1459-1465
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    • 2013
  • Two in vitro experiments were conducted to evaluate the effect of methylcellulose (MC) on i) bacterial detachment from rice straw as well as ii) inhibition of bacterial attachment and fiber digestibility. To evaluate the effect of MC on fibrolytic bacterial detachment (Exp 1), in vitro bacterial cultures with 0.1% (w/v) MC solution were compared with cultures without MC after 8 h incubation. The effect of MC on inhibition of bacterial attachment was determined by comparing with real-time PCR the populations of F. succinogenes, R. flavefaciens and R. albus established on rice straw pre-treated with 0.1% MC with those on untreated straw after incubation for 0, 6 and 12 h (Exp 2). The major fibrolytic bacterial attachment on rice straw showed significantly lower populations with either the addition of MC to the culture or pre-treated rice straw compared to controls (p<0.05). Also, the digestibility of rice straw with MC was significantly lower compared with control (p<0.05). The F. succinogenes population did not show detachment from rice straw, but showed an inhibition of attachment and proliferation on rice straw in accordance with a decrease of fiber digestion. The detachments of Ruminococcus species co-existed preventing the proliferations with subsequent reduction of fiber degradation by MC during the incubation. Their detachments were induced from stable colonization as well as the initial adhesion on rice straw by MC in in vitro ruminal fermentation. Furthermore, the detachment of R. albus was more sensitive to MC than was R. flavefaciens. These results showed the certain evidence that attachment of major fibrolytic bacteria had an effect on fiber digestion in the rumen, and each of fibrolytic bacteria, F. succinogenes, R. flavefaciens and R. albus had a specific mechanism of attachment and detachment to fiber.

Preparation of Hypoallergenic Whey Protein Hydrolysate by a Mixture of Alcalase and Prozyme and Evaluation of Its Digestibility and Immunoregulatory Properties

  • Jiyeon Yang;Se Kyung Lee;Young Suk Kim;Hyung Joo Suh;Yejin Ahn
    • Food Science of Animal Resources
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    • v.43 no.4
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    • pp.594-611
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    • 2023
  • Whey protein (WP) has nutritional value, but the presence of β-lactoglobulin (β-LG) and α-lactalbumin (α-LA) cause allergic reactions. In this study, hypoallergenic whey protein hydrolyate (HWPH) was prepared by decomposing β-LG and α-LA of WP using exo- and endo-type proteases. The enzyme mixing ratio and reaction conditions were optimized using response surface methodology (RSM). Degradation of α-LA and β-LG was confirmed through gel electrophoresis, and digestion, and absorption rate, and immunostimulatory response were measured using in vitro and in vivo systems. Through RSM analysis, the optimal hydrolysis conditions for degradation of α-LA and β-LG included a 1:1 mixture of Alcalase and Prozyme reacted for 10 h at a 1.0% enzyme concentration relative to substrate. The molecular weight of HWPH was <5 kDa, and leucine was the prominent free amino acid. Both in vitro and in vivo tests showed that digestibility and intestinal permeability were higher in HWPH than in WP. In BALB/c mice, as compared to WP, HWPH reduced allergic reactions by inducing elevated Type 1/Type 2 helper T cell ratio in the blood, splenocytes, and small intestine. Thus, HWPH may be utilized in a variety of low allergenicity products intended for infants, adults, and the elderly.

Combination of Grapefruit and Rosemary Extracts Has Skin Protective Effect through MMPs, MAPKs, and the NF-κB Signaling Pathway In Vitro and In Vivo UVB-exposed Model

  • Yoon, Yeo-Cho;Choi, Hee-Jeong;Park, Ji-Hyun;Diniyah, Nurud;Shin, Hyun-A;Kim, Mi-Yeon
    • Korean Journal of Plant Resources
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    • v.32 no.6
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    • pp.633-643
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    • 2019
  • Long-term ultraviolet (UV) exposure accelerates the phenomenon of skin photo-aging by activating collagenase and elastase. In this study, we aimed to investigate the effects of a combination of grapefruit and rosemary extracts (cG&Re) on UVB-irradiated damage in HaCaT cells and dorsal mouse skin. In HaCaT cells, cG&Re recovered UVB-reduced cell viability and inhibited protein expression of mitogen-activated protein kinases (MAPKs), such as extracellular signal-regulated kinases (p-Erk), c-Jun N-terminal kinases (p-JNK), and a class of MAPKs (p-P38). Also, cG&Re suppressed UVB-induced collagen and elastin degradation by decreasing matrix metalloproteinases (MMPs) and nuclear factor kappa light chain enhancer of activated B cells (NF-κB) expression, which is a transcription factor. Similar results were observed in dorsal mouse skin. Taken together, our data indicate that cG&Re prevent UVB-induced skin photo-aging due to collagen/elastin degradation via activation of MAPKs, MMPs, and the NF-κB signaling pathway in vitro and in vivo.