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Production of bioactive ginsenoside Rg3(S) and compound K using recombinant Lactococcus lactis

  • Li, Ling;Lee, Soo Jin;Yuan, Qiu Ping;Im, Wan Taek;Kim, Sun Chang;Han, Nam Soo
    • Journal of Ginseng Research
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    • v.42 no.4
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    • pp.412-418
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    • 2018
  • Background: Ginsenoside Rg3(S) and compound K (C-K) are pharmacologically active components of ginseng that promote human health and improve quality of life. The aim of this study was to produce Rg3(S) and C-K from ginseng extract using recombinant Lactococcus lactis. Methods: L. lactis subsp. cremoris NZ9000 (L. lactis NZ9000), which harbors ${\beta}$-glucosidase genes (BglPm and BglBX10) from Paenibacillus mucilaginosus and Flavobacterium johnsoniae, respectively, was reacted with ginseng extract (protopanaxadiol-type ginsenoside mixture). Results: Crude enzyme activity of BglBX10 values comprised 0.001 unit/mL and 0.003 unit/mL in uninduced and induced preparations, respectively. When whole cells of L. lactis harboring pNZBglBX10 were treated with ginseng extract, after permeabilization of cells by xylene, Rb1 and Rd were converted into Rg3(S) with a conversion yield of 61%. C-K was also produced by sequential reactions of the permeabilized cells harboring each pNZBgl and pNZBglBX10, resulting in a 70% maximum conversion yield. Conclusion: This study demonstrates that the lactic acid bacteria having specific ${\beta}$-glucosidase activity can be used to enhance the health benefits of Panax ginseng in either fermented foods or bioconversion processes.

Purification and Characterization of a Novel Salt-tolerant Protease Produced by Saccharomyces sp. B101 Isolated from Baker's Dough Yeast

  • Hwang, Joo-Yeon;Kim, Sang-Moo;Heo, Seok;Kim, Cheon-Jei;Lee, Chi-Ho;Lee, Si-Kyung
    • Food Science and Biotechnology
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    • v.17 no.4
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    • pp.766-771
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    • 2008
  • The proteolytic enzyme from Saccharomyces sp. B101 was purified to homogeneity by ammonium sulfate fractionation, ultrafiltration, diethyl aminoethyl (DEAE)-Sephadex A-50 ion-exchange chromatography, and Sephadex G-100 gel filtration chromatography from the culture supernatant of Saccharomyces sp. B101. The specific activity and the purification fold of the purified enzyme were 4,688.9 unit/mg and 18, respectively. The molecular weight of the purified enzyme was estimated to be 33 kDa by sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The optimum pH and temperature for the enzyme activity were pH 8.5 and $30^{\circ}C$, respectively. The enzyme activity was relatively stable in the pH range of 6.5-8.5 at below $35^{\circ}C$. The salt-tolerance and stability for the enzyme activity were relatively stable even at NaCl concentrations of 10 and 15%. The activity of enzyme was inhibited by $Ag^{2+}$ and $Fe^{2+}$, and activated by $Mn^{2+}$. In addition, the enzyme activity was potently inhibited by ethylenediaminetetraacetic acid (EDTA) and phenylmethyl sulfonylfluoride (PMSF). Based on these findings we concluded that the purified enzyme was a serine protease. Km and Vmax values for hammastein milk casein were 1.02 mg/mL and 278.38 unit/mL, respectively.

Genetic alterations in Wnt family of genes and their putative association with head and neck squamous cell carcinoma

  • Aditya, Jain;Smiline Girija, A.S.;Paramasivam, A.;Priyadharsini, J. Vijayashree
    • Genomics & Informatics
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    • v.19 no.1
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    • pp.5.1-5.11
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    • 2021
  • Head and neck squamous cell carcinoma (HNSCC) is the most frequent type of head and neck cancer that usually arises from the mucosal surfaces of several organs including nasal cavity, paranasal sinuses, oral cavity, tongue, pharynx, and larynx. The Wnt signaling pathway is a crucial mechanism for cellular maintenance and development. It regulates cell cycle progression, apoptosis, proliferation, migration, and differentiation. Dysregulation of this pathway correlates with oncogenesis in various tissues including breast, colon, pancreatic as well as head and neck cancers. The present study aims to assess the gene alterations in the Wnt family of genes so as to derive an association with HNSCC. Computational approaches have been utilized for the identification of gene alterations in the Wnt family of genes. Several databases such as cBioportal, STRING, and UALCAN were used for the purpose. The frequency of alteration was high in case of Wnt family member 11 (5%). Gene amplification, deep deletions, missense and truncating mutations were observed in HNSCC patients. There was a marked difference in the gene expression profile of WNT11 between grades as well as normal samples. The survival probability measured using the Kaplan-Meier curve also presented with a significant difference among male and female subjects experiencing a low/medium level expression. The female patients showed less survival probability when compared to the male subjects. This provides the prognostic significance of the WNT11 gene in HNSCC. Taken together, the present study provides clues on the possible association of WNT11 gene alterations with HNSCC, which has to be further validated using experimental approaches.

Dysregulation of Cannabinoid CB1 Receptor Expression in Subcutaneous Adipocytes of Obese Individuals

  • Lee, Yong-Ho;Tharp, William G.;Dixon, Anne E.;Spaulding, Laurie;Trost, Susanne;Nair, Saraswathy;Permana, Paska A.;Pratley, Ridhard E.
    • Animal cells and systems
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    • v.13 no.4
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    • pp.371-379
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    • 2009
  • The endocannabinoid system (ECS) plays a key role in the regulation of appetite, body weight and metabolism. We undertook the present study to further clarify the regulation of the cannabinoid CB1 receptor (CB1, CNR1) in human adipose tissue in obesity. CB1 receptor mRNA expression was ~1.6-fold (p<0.004) and 1.9-fold higher (P<0.05) in subcutaneous adipocytes from obese compared to non-obese subjects in microarray and quantitative real-time PCR studies, respectively. Higher CB1 receptor mRNA expression levels in both adipose tissue (~1.2 fold, P<0.05) and adipocytes (~2 fold, P<0.01) were observed in samples from visceral compared to subcutaneous depots collected from 22 obese individuals. Immunofluorescence confocal microscopy demonstrated the presence of CB1 receptor on adipocytes and also adipose tissue macrophages. These data indicate that adipocyte CB1 receptor is up-regulated in human obesity and visceral adipose tissue and also suggest a potential role for the ECS in modulating immune/inflammation as well as fat metabolism in adipose tissue.

Effects of Dietary Carbohydrase Enzyme Complex and Microbial Phytase Supplementation on Productivity and Nutrient Digestibility in Growing Pigs (탄수화물 분해 복합효소제와 미생물 파이테이즈의 첨가가 육성돈의 생산성 및 영양소 소화율에 미치는 영향)

  • Shim, Y.H.;Chae, B.J.;Lee, J.H.
    • Journal of Animal Science and Technology
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    • v.45 no.4
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    • pp.569-576
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    • 2003
  • An experiment was conducted to investigate the effect of microbial phytase (Natuphos$^{\circledR}$) supplementation, individually and in combination with carbohydrase enzyme complex (composed of enzymes targeted to SBM dietary components such as $\alpha$-galactosides and galactomannans; ENDO-POWER$^{\circledR}$) to corn-soy basis diet with low nutrient levels on growth performance and nutrient digestibility of growing pigs. A total of 48 crossbred weaned pigs (Landrace${\times}$Yorkshire${\times}$Duroc), 29.1$\pm$0.14 kg of initial body weight, were randomly allotted to four dietary treatments, based on weight and age, according to a Randomized Complete Block Design. There were three pens per treatment and 4 pigs per pen. The dietary treatments were 1) CON (control diet with 3,380 kcal/kg of metabolizable energy, 18.96% of crude protein, 1.10% of lysine, 0.75% of calcium and 0.35% of available phosphorus), 2) LP+NTPS (CON diet with 0.15% unit lower available P levels+0.1% phytase (500 FTU/kg; Natuphos$^{\circledR}$)), 3) LEL+ENP (CON diet with 3.0% unit lower ME and lysine levels + 0.1% carbohydrase enzyme complex (ENDO-POWER$^{\circledR}$), and 4) LPEL+ENZ (CON diet with 0.15% unit lower available P levels and 3.0% unit lower ME and lysine levels+0.1% ENDO-POWER$^{\circledR}$ and 0.1% Natuphos$^{\circledR}$ (500 FTU/kg). There was no significant difference (p〉0.05) in average daily gain (ADG), average daily feed intake (ADFI) and feed conversion ratio (FCR) among dietary treatments during the whole experimental period (0 to 4 weeks). Apparent digestibility of gross energy was greater in LP+NTPS and LPEL+ENZ groups than in the LEL+ENP (p<0.05). Apparent digestibility of phosphorus was greater in LP+NTPS than in LEL+ENP (p<0.05). Dry matter excretion was lowest in LPEL+ENZ and phosphorus excretion was lowest in LP+NTPS (p<0.05). Overall, pigs fed on LPEL+ENZ group tended to have better nutrient digestibility (dry matter, gross energy, crude protein and phosphorus) than pigs fed on control group. All dietary enzyme treatment groups showed lower feed cost/body weight gain of pigs than control group. In conclusion, the results from the present study suggest that the simultaneous inclusion of phytase and carbohydrase enzyme complex to diets is advantageous with respect to reducing nutrient excretion of growing pigs and may contribute to increased economic return when added to corn-soy based growing pig diets.

Effect of Agariemycetes Supplementation on Egg Quality and Fecal Noxious Gas Concentration in Laying Hens (사료내 Agariemycetes 첨가 급여가 산란계의 계란 품질 및 분내 유해 가스 농도에 미치는 영향)

  • Jung, Ji-Hong;Kim, Hyo-Jin;Wang, Jian Ping;Kim, Chae-Hyun;Jung, Cheong-Hwan;Kim, In-Ho
    • Korean Journal of Poultry Science
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    • v.37 no.2
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    • pp.145-150
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    • 2010
  • This study was conducted to evaluate effects of dietary Agariemycetes on egg quality and fecal noxious gas concentration in laying hens. A total of two hundred forty laying hens were randomly allocated to four treatments with five replicates of twelve chicks per pen. The experiment lasted 42 days with a 7-day adjustment period. Dietary treatments were as follows: 1) CON (basal diet), 2) A1 (basal diet + Agariemycetes 0.1%), 3) A3 (basal diet + Agariemycetes 0.3%) and 4) A5 (basal diet + Agariemycetes 0.5%). There was no significant difference in egg production and egg weight between CON and Agariemycetes treatments. During the second week of the experiment, eggshell breaking strength was higher in A3 and A5 treatments than that in CON treatment (P<0.05). Besides, eggshell breaking strength in P5 treatment was higher when compared with that in CON treatment (P<0.05) in the sixth week. During the second week of the experiment, eggshell thickness was higher in Agariemycetes treatments than CON treatment (P<0.05). Also, A5 treatment was higher than other treatments in the fourth week of the experiment (P<0.05). At the end of the second week, yolk color unit in A5 treatment was lower compared with CON treatment (P<0.05). During the fourth week of the experiment, CON treatment was higher than A3 treatment (P<0.05). During the second week of the experiment, Haugh unit in A3 treatment was greater than that in CON treatment (P<0.05). At the end of six weeks, Haugh unit in CON treatment was lower than other treatments (P<0.05). Fecal ammonia and hydrogen sulfide were increased in Agariemycetes treatments when compared with CON treatment (P<0.05). Compared with CON and A5 treatments, fecal mercaptans was higher than in other treatments in first of the experiment (P<0.05). In conclusion, this study indicated that the addition of Agariemycetes in the diet improved the egg quality and decreased the feacal noxious gas concentration in laying hens.

The Development of Differentiating Method between Fresh and Frozen Beef by Using the Mitochondrial Malate Dehydrogenase Activity (Mitochondrial Malate Dehydrogenase 활성을 이용한 냉장우육과 냉동우육의 판별법 개발)

  • Han, Kyu-Ho;Kim, Nam-Kyu;Lee, Si-Kyung;Cho, Jin-Kook;Choi, Kang-Duk;Jeons, You-Jin;Lee, Chi-Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.10
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    • pp.1599-1605
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    • 2005
  • The object of this study is to develop the method for differentiating fresh meat from frozen meat by using the measurement of the mitochondrial malate dehydrogenase in the Korean native cattle. The principle of this experiment is based on the fact that the enzyme proteins associated with mitochondrial membrane could be released by freezing. The methods of differentiating fresh meat from thawed, frozen meat were studied by measurements of mitochondrial malate dehydrogenase activity of meat press juice. Fresh and frozen beef were stored at 4, -4, -18 and -77$^{\circ}C$ for 15-day storage period. A meat press machine using air pressure was manufactured especially for these experiments, and sufficient amount of drip (about 0.15 mL/g) from 1.5 g of beef sample was efficiently obtained under a pressure of 8 kg/$cm^{2}$ generated by the meat pressing machine. The mitochondrial malate dehydrogenase activities of frozen meat drip i년ices stored at -18 and -77$^{\circ}C$ were significantly higher than those of fresh and frozen meat samples at -4$^{\circ}C$ (p < 0.05) during 10-min reaction period. However, the enzyme activities of the frozen meat drip juices (-18 and -77$^{\circ}C$) disappeared after 5 minutes of the reaction, which was not observed from the fresh and -4$^{\circ}C$ frozen meats. The enzyme activity maintained until 12 minutes for the fresh and -4$^{\circ}C$ frozen meats. From these results, the mitochondrial malate dehydrogenase could be considered as an indicator to differentiate fresh beef from frozen one.

Nutrition Practice to Alleviate the Adverse Effects of Stress on Laying Performance, Metabolic Profile and Egg Quality in Peak Producing Hens: II. The Probiotic Supplementation

  • Hayirli, Armagan;Esenbuga, N.;Macit, M.;Yoruk, M.A.;Yildiz, A.;Karaca, H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.12
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    • pp.1752-1760
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    • 2005
  • In this experiment, the effects of cage density (CD) and probiotic supplementation (PS) on laying performance, metabolic profile, and egg quality in peak-producing hens were evaluated. After blocking according to the cage location, Lohman layers (n = 180, 46 wks of age) were allocated randomly to two levels of CD (540 vs. 360 $cm^2$/hen) and three levels of PS (0, 0.15, and 0.30%). Probiotic contained Enterococcus faecium culture (10${\times}$10$^9$ cfu/g). Egg production (EP) and feed consumption (FC) were measured daily; egg weight (EW) was measured bi-weekly; BW was measured before and after the experiment; and blood samples were obtained at the end of the experiment. The data were analyzed using two-way ANOVA. Increasing CD decreased FC (125.0 vs. 120.8 g/d, p<0.0001) and FCR (1.93 vs. 1.87, p<0.0001) and did not alter EP, EW, and BW. Increasing level of PS linearly decreased FC (p<0.02) and FCR (p<0.006). Averages were 123.9, 123.2, and 121.6 g/d for FC and 1.91, 1.92, and 1.86 for FCR in hens supplemented with 0, 0.15, and 0.30% probiotic, respectively. Hens placed in high-density cages had greater serum corticosterone concentration than hens placed in normal-density cages (12.8 vs. 11.3 $\mu$g/dL, p<0.04); CD did not affect concentrations of other metabolites. Increasing level of PS linearly increased serum glucose, albumin, and creatine concentrations and quadratically increased total protein, globulin, Ca, and P concentrations. Average concentrations (mg/dL) were 260, 297, and 305 for glucose; 6.28, 8.09, and 7.58 for total protein; 1.98, 2.48, and 2.38 for albumin; 4.30, 5.62, and 5.19 for globulin; 0.40, 0.52, and 0.54 for creatine; 16.0, 16.5, and 16.3 for Ca; and 6.27, 8.14, and 7.17 for P in hens supplemented with 0, 0.15, and 0.30% probiotic, respectively. There was no effect of CD on egg quality. Increasing level of PS linearly improved yolk color (YC) and quadratically increased albumen index (AI) and Haugh unit (HU). The mean values were 9.67, 9.75, and 10.58 for YC; 8.94, 6.93, and 8.72% for AI; and 85.6, 74.9, and 82.9 for HU for hens supplemented with 0, 0.15, and 0.30% probiotic, respectively. There was also CD by PS effect on FC, EP, and serum glucose, total protein, albumin, globulin, creatine, Ca and P concentrations. In conclusion, increased CD partially depressed laying performance and caused stress. Probiotic supplementation improved laying performance and metabolic profile. It also partially alleviated the adverse effects of stress resulting from increased caging density.

Effects of Weaning and Spatial Enrichment on Behavior of Turkish Saanen Goat Kids

  • Tolu, Cemil;Gokturk, Semra;Savas, Turker
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.6
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    • pp.879-886
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    • 2016
  • As is in all economic activities, the highest yield per unit area is the main goal in animal production, while addressing the temperamental needs of animals often is ignored. Animal welfare is not only an ethical fact; it also has an economic value. Spatial environmental enrichment contributes positively to animal welfare by addressing their behavioral and mental requirements. The present study was conducted to determine the effects of weaning and spatial environmental arrangements on behaviors of goat-kids. Experimental groups were arranged in structured and unstructured spatial environments. Roughage feeder, semi-automatic concentrate feeder, bunk, bridge, and wood block were placed in the structured environment. No equipment was placed in the unstructured environment and paddock sides were enclosed with an iron sheet to prevent bipedal stance and to provide environmental isolation. In the study 10 male and 10 female Turkish Saanen goat kids were used in each group. Spatial environmental arrangements did not have significant impacts on the growth performance of kids (p>0.05). All objects in the structured group were accepted by the kids. Average use ratios of roughage feeder, semi-automatic concentrate feeder, bunk, bridge and wood block were observed as 19.3%, 14.0%, 12.6%, 3.8%, and 0.7%, respectively. There were significant differences between before- and after-weaning in use of all objects except for underneath bridge ($p{\leq}0.05$). Concentrate feed consumption, locomotion, and resting behaviors in kids showed significant differences by structural group and growth period. Roughage consumption was similar between groups, while it differed by growth period ($p{\leq}0.05$). Interaction frequency was significantly higher in structured group (p = 0.0023). Playing behavior significantly differentiated based on the growth period rather than on groups ($p{\leq}0.05$). Playing behavior significantly decreased after weaning. Abnormal oral activity was significantly higher in the structured group before weaning ($p{\leq}0.05$). Despite there being no installations facilitating climbing and bipedal stance, the kids of the unstructured group were able to exhibit 1/3 as much bipedal stance behavior as the kids of the structured group through leaning over slippery paddock wall or over their groupmates. Bipedal stance behavior of unstructured group was similar before and after weaning, while bipedal stance behavior before weaning was about 2 times that of after weaning in structured group. It was concluded that unstructured environmental arrangement limited the behavior repertoire of the goat kids.

Effect of Stocking Density on the Feather Condition, Egg Quality, Blood Parameters and Corticosterone Concentration of Laying Hens in Conventional Cage (케이지 사육시 사육밀도가 산란계의 깃털손상도, 계란품질, 혈액성상 및 스트레스 호르몬 농도에 미치는 영향)

  • Son, Ji Seon;Kim, Chan Ho;Kang, Hwan Ku;Kim, Hyun Soo;Jeon, Jin Joo;Hong, Eui Chul;Kang, Bo Seok
    • Korean Journal of Poultry Science
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    • v.47 no.2
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    • pp.83-93
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    • 2020
  • The purpose of this study was to evaluate the effects of stocking density on feather condition, egg quality, blood parameters, and corticosterone levels of laying hens reared in conventional cages. A total of 500, 28 weeks-old Hy-Line Brown laying hens were evenly assigned to two stocking densities (750 and 500 ㎠/bird). There were 50 replicates of each treatment, each consisting of 750 ㎠/birds (n=6) or 500 ㎠/bird (n=4) birds in a cage, respectively. We observed an increase in the laying period of, the group that was assigned 500 ㎠/bird, however, this group experienced significant feather damage when compared with the group allotted 750 ㎠ bird. There was no effect of stocking densities on egg quality. These results suggest that the 500 ㎠/bird stocking density in the conventional cage can negatively affect feather condition and cause the birds to experience more stress than the group allotted 750 ㎠/bird.