• Title/Summary/Keyword: crude extract

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Fruit and vegetable discards preserved with sodium metabisulfite as a high-moisture ingredient in total mixed ration for ruminants: effect on in vitro ruminal fermentation and in vivo metabolism

  • Ahmadi, Farhad;Lee, Won Hee;Oh, Young-Kyoon;Park, Keunkyu;Kwak, Wan Sup
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.3
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    • pp.446-455
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    • 2020
  • Objective: Our recent series of laboratory- and large-scale experiments confirmed that under aerobic and anaerobic conditions, sodium metabisulfite (SMB) was effective in preserving nutrients and antioxidant capacity of highly perishable fruit and vegetable discards (FVD). Hence, the purpose of this study was to examine how partial inclusion of SMB-treated FVD in total mixed ration (TMR) influences in vitro ruminal fermentation, whole-tract digestibility, nitrogen metabolism, blood metabolites, and voluntary feed intake of sheep. Methods: The FVD were mixed thoroughly with 6 g SMB/kg wet biomass and kept outdoors under aerobic conditions for 7 days. Four TMRs including four levels of SMB-treated FVD (as-fed basis) at 0%, 10%, 20%, and 30% (equaling to 0%, 1.9%, 3.8%, and 5.7% on dry matter basis, respectively), were prepared as replacement for corn grain. The ruminal fermentation metabolites were studied using an in vitro gas production test. Four mature male Corriedale sheep were assigned at random to the 4 diets for two separate sub-experiments; i) digestibility trial with four 21-d periods, and ii) voluntary feed intake trial with four 28-d periods. Results: Inclusion of SMB-treated FVD in the TMR tended to quadratically increase partitioning factor. No effect was seen on total-tract digestibility of organic matter, ether extract, crude protein, and acid detergent fiber, except for neutral detergent fiber digestibility that tended to linearly increase with increasing SMB-treated FVD in the TMR. The progressive increase of FVD preserved with SMB in the diet had no effect on nitrogen metabolism. Treatment had no effect on serum antioxidant capacity and blood metabolites assayed. Voluntary feed intake was not impaired by inclusion of SMB-treated FVD in the TMR. Conclusion: It appears that FVD preserved with SMB can be safely incorporated into TMR as replacement of corn grain without impairment of nutrient metabolism and feed intake.

Production and Separation of Anti-hypertensive Peptide during Chunggugjang Fermentation with Bacillus subtilis CH-1023 (청국장 발효과정 중 항고혈압성 peptide의 생산 및 분리)

  • Cha, Woen-Suep;Bok, Su-Kyung;Kim, Myoung-Uk;Chun, Sung-Sook;Choi, Ung-Kyu;Cho, Young-Je
    • Applied Biological Chemistry
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    • v.43 no.4
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    • pp.247-252
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    • 2000
  • As functionality investigation of Korean traditional soybean fermentation foods, an antihypertensive peptide was separated during Chunggugjang fermentation by Bacillus subtilis CH-1023 and investigated inhibitory effect against angiotensin converting enzyme. After incubation at $20^{\circ}C,\;30^{\circ}C,\;40^{\circ}C,\;50^{\circ}C,\;60^{\circ}C$ for the $0{\sim}72$ hrs, protein content, protease activity and angiotensin converting enzyme inhibitory rate were determined. The protein content and protease activity were increased and reached maximum at 60 hrs fermentation with $40^{\circ}C$ and decreased after the 60 hrs fermentation. The optimum condition for antihypertensive peptide from Chunggugjang was appeared for 60 hrs at $40^{\circ}C$. Crude extract of Chunggugjang was partially purified by Amicon YM-3 membrane filtration and Sephadex G-10, G-25 gel filtration. The purified peptide showed inhibitory rate of 94.3% with 0.5 mg peptide content. The most prominent amino acid composition of the peptide from Chunggugjang was alanine, followed by phenylalanine, histidine.

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Inhibition Effect against the Rat Blood Plasma Oxidation of the $Makgeolli$ ($Takju$) Korean Rice Wine (막걸리(탁주)의 쥐 혈장 과산화물 생성 억제효과)

  • Wang, Seung-Jin;Lee, Hyoung-Jae;Cho, Jeong-Yong;Jang, Mi-Young;Park, Keun-Hyung;Moon, Jae-Hak
    • Food Science and Preservation
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    • v.19 no.1
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    • pp.116-122
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    • 2012
  • The antioxidant activities of $makgeolli$ and other alcoholic beverages were compared. Based on the same volume (70 ${\mu}L$ eq.) of the alcoholic beverages, the 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid ammonium salt) (ABTS+) radical-scavenging activities were as follows: whisky > $makgeolli$ crude extract (MCE) > rice wine (RW) > clarified $makgeolli$ (CM) > soju. Based on the same alcohol concentration (6%) of the alcoholic beverages, however, $makgeolli$ showed the highest activity. In addition, based on the same volume (70 ${\mu}L$ eq.), the inhibition effects against the formation of cholesteryl ester hydroperoxide (CE-OOH) were as follows: soju > whisky > RW > MCE > CM. Based on the same alcohol concentration (6%), however, the inhibition effects against the formation of CE-OOH were as follows: RW > MCE > soju > whisky > CM. Therefore, it was suggested that $makgeolli$ may contain radical-scavenging- and metal-ion-chelate-type antioxidants and may increase the antioxidant activity in the blood.

Antioxidant and Antimicrobial Effects of Solvent Fractions from Smilax china L. Leaves (청미래덩굴 잎(Smilax china L.) 용매 분획물의 항산화 및 항균 효과)

  • Kim, Joo-Young;Seoung, Gi-Un;Chung, Shin-Kyo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.10
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    • pp.1614-1618
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    • 2014
  • To utilize Smilax china L. leaves as a natural preservatives, solvent fractions from crude methanol extract were prepared and investigated their antioxidant and antimicrobial activities against Staphylococcus aureus. Antioxidant activities were examined by 1,1-diphenyl-1-picrylhydrazyl radical scavenging and ferric ion reducing antioxidant power assays. Ethyl acetate fraction from Smilax china L. leaves exhibited the strongest antioxidant and antimicrobial activities among the fractions (P<0.05), as well as the highest total phenolic and total flavonoid contents. Caffeic acid, ferulic acid, quercetin, and kaempferol contents in the ethyl acetate fraction from Smilax china L. leaves were determined by TLC and HPLC. In aqueous phase, as well as the n-butanol and n-hexane fractions, quercetin, ferulic acid, and kaempferol were detected in small amounts. Ferulic acid, which showed the highest content, is the major active compound from Smilax china L. leaves.

Effect of Jeseupwiryeongtang-gagam on Atopic Dermatitis-like Skin NC/Nga Mouse Induced by Dermatophagoides Farinae Crude Extract (제습위령탕가감이 집 먼지 진드기 추출물로 유도된 NC/Nga mouse의 아토피 피부염에 미치는 영향)

  • Na, Dong-Gyu;Han, Jae-Kyung;Kim, Yun-Hee
    • The Journal of Pediatrics of Korean Medicine
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    • v.22 no.2
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    • pp.81-114
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    • 2008
  • Objectives : The purpose of this study is to investigate the effect of Jeseupwiryeongtang-gagam(JWTG) on atopic dermatitis by in vivo experiment using NC/Nga atopic dermatitis mouse, which has histological and clinical similarities to the atopic dermatitis of human. Methods : To investigate the effect of JWTG on AD, we evaluated atopic dermatitis-like skin lesions by clinical skin index and analyzed immunological parameters in peripheral blood mononuclear cells(PBMCs), splenocytes, draining lymph node(DLN) and performed skin histology in ears and dorsal skin of atopic dermatitis-like skin NC/Nga mouse in vivo. Results and Conclusions : In vivo, clinical skin severity score were significantly lower in JWTG group than control group. IgE, IL-6, $TNF-{\alpha}$, IgM, IgG2a and IgG2b levels in serum were decreased remarkably in JWTG group than control group and $IFN-{\gamma}$ production, secreted in Th1 cell were increased by JWTG. After experiment ended, we analyzed immunological cells ($CD3^+$, $CD19^+$, $CD4^+$, $CD8^+$, $CD3^+$$CD69^+$, $CD4^+$$CD25^+$ and $CD49b^+$) by flow cytometry. It resulted that total absolute number of $CD3^+$, $CD19^+$, $CD4^+$ and $CD8^+$ cells were recovered as normal and $CD3^+$$CD69^+$ were decreased significantly compared with control group in isolated DLN and PBMCs from NC/Nga mouse and total absolute number of $Gr-1^+$, $CD11b^+$ and $CD3^+$ in dorsal skin of NC/Nga mouse were decreased by JWTG. We analyzed ear, DLN, and neck-back skin after biopsy and dyeing by hematoxyline/eosin(H&E) and toluidine staining (mast cells marker) and obtained results that JWTG were effective to histological symptoms (dermal and epidermal thickening, hyperkeratosis and inflammatory cell infiltration). Ear thickness was decreased significantly than the control group and the size of inflammatory lymphocytes cells(ILC) and plasma cells(PC) in DLN were also decreased.

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Seroprevalence of Toxoplasmosis with ELISA and Rapid Diagnostic Test among Residents in Gyodong-do, Inchon city, Korea: A Four-Year Follow-up

  • Kim, Yeong Hoon;Lee, Ji hoo;Ahn, Seong kyu;Kim, Tong-Soo;Hong, Sung-Jong;Chong, Chom-Kyu;Ahn, Hye-Jin;Nam, Ho-Woo
    • Parasites, Hosts and Diseases
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    • v.55 no.3
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    • pp.247-254
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    • 2017
  • ELISA has been used for the diagnosis of toxoplasmosis, but it is being gradually replaced by a rapid diagnostic test (RDT). We compared and analyzed ELISA and RDT results using the sera collected during 4 consecutive years from residents of Gyodong-do (Island), Incheon-city, Korea. Sera from 921, 993, 940, and 838 adult residents were collected on a yearly basis (2010-2013). ELISA was performed by using a crude extract of T. gondii RH strain antigen and IgG/IgM RDT mounted with recombinant fragment of major surface antigen (SAG1), GST-linker-SAG1A, were applied to the sera. Comparison between groups was analyzed by the Student's t-test. The positive seroprevalence surged from 14.7% (135/921, 2010), 23.1% (231/993, 2011), 23.6% (222/940, 2012), and 32.1% (269/838, 2013) by ELISA. In contrast, RDT showed a more moderate increasing trend from 21.7% (200/921, 2010), 25.5% (253/993, 2011), 28.9% (272/940, 2012) and 33.1% (277/838, 2013). Discrepancies between ELISA and RDT were noted near the cut-off value. At the OD 0.15-0.24 range, RDT could detect 16.1% (169/1051) more positives, which suggests an early or acute toxoplasmosis, but at the OD 0.25-0.34 range, ELISA could detect 35.9% (92/256) more positives of possible chronic infections. Over the OD > 0.35 ELISA and RDT agreed in the majority of the cases. This surge in seroprevalence may be caused by the organic agriculture in addition to eating behavior or increase in pets among Koreans. These facts may be applied on a full-scale national survey using RDT to supplement ELISA to define the characteristics of the infection.

Pathogene Resistance of cotton GST cDNA in Transgenic Scrophularia buergeriana Misrule (목화 Glutathione S-Transferase (GST) 유전자로 형질 전환된 현삼의 내병성 특성)

  • 강원희;임정대;이성호;유창연
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.6
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    • pp.297-304
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    • 2001
  • Scrophularia buergeriana Misrule has been contaminated with various pathogens in condition of field and storage period. This study was carried out for production of multiple stress resistance plant containing disease resistance that CGST gene expressed in transgenic Scrophularia buergeriana Misrule genome. Glutathione S-Transferases (GSTs) detoxify endobiotic and xenobiotic compounds by covalent linking of tripeptide glutathione to hydrophobic substrate. GST enzymes have been identified and characterized in insects, bacteria, and many plant species. A cDNA clone of GST was introduced into Scrophularia buergeriana Miquel by transformation with Agrobacterium tumefaciences. In coporation of the CGST gene into S. buergeriana Misrule was confirmed by PCR analysis of genomic DNA. Influence of exposure to darkness on the regeneration potential and transformation frequence were assessed. The activity of GST in transgenic plants was two times higher than that of non-transgenic plants. As a result of anti-microbe assays, the crude extract protein of transgenic plants showed the antimicrobial effects higher than control plants.

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Purification and Biochemical Characteristics of a 45 kDa Fibrinolytic Enzyme from a Halophile (호염성균 유래 45 kDa 혈전용해효소의 순수분리와 생화학적 특성)

  • Kim, Do-Hyoung;Park, Jeong-Uck;Seo, Min-Jeong;Kim, Min-Jeong;Lee, Hye-Hyeon;Choi, Yung-Hyun;Joo, Woo-Hong;Jeong, Yong-Kee
    • Journal of Life Science
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    • v.20 no.2
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    • pp.183-189
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    • 2010
  • A fibrinolytic enzyme producing Bacillus sp. J-19 was isolated from the popular Korean seasoning, pickled anchovy. The fibrinolytic enzyme was purified to homogeneity by chromatographic methods including ethanol precipitation and gel-filtration using Sephadex G-50. Compared to the crude enzyme extract, the specific activity of the enzyme increased 1021-fold with a recovery of 23%. The purified enzyme was estimated to be approximately 45 kDa by SDS-PAGE. Especially, the amidolytic activity in the presence of the synthetic substrate for serine protease (H-D-Ile-Pro-Arg-pNA, S-2288) represented approximately 17 U/mg. In addition, more than the 60% activity of the 45 kDa fibrinolytic activity was maintained in the presence of up to 30% (w/v) sodium chloride. These findings could provide a unique fibrinolytic enzyme, leading to a potential thrombolytic agent.

Use of the Synthetic Gene Encoding the Truncated Human Rotavirus VP8* Protein in Escherichia coli for Production of Vaccine Candidates or Development of Diagnostic Antibodies (합성 유전자를 이용하여 Escherichia coli에서 백신 후보의 생산 혹은 진단용 항체의 개발을 위한 인간 rotavirus VP8* 부분 단백질의 발현)

  • Kim, Sang-Rae;Lee, Bheong-Uk
    • Journal of Life Science
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    • v.28 no.4
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    • pp.478-482
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    • 2018
  • Human rotavirus is a causative agent of acute diarrhea among children. The artificial gene encoding the truncated $VP8^*$ protein of human rotavirus A (serotype 1 strain WA) was synthesized according to the Escherichia coli codon preference. The synthetic $VP8^*$ gene also possessed the NdeI and HindIII restriction sites for the convenient in-frame cloning for translation and a 6-histidine tag at C-terminus for Ni+ affinity purification. Molecular weight of the truncated $VP8^*$ protein deduced from the nucleotide sequences of the artificial gene was a 19.7-kDa. This synthetic $VP8^*$ DNA fragment was inserted into the pT7-7 expression vector and transformed into E. coli BL21 (DE3). Transformants harboring the synthetic gene encoding the $VP8^*$ protein was induced by supplement of a final concentration of 0.05 mM ITPG at $20^{\circ}C$. Protein crude extract from the E. coli transformants was subjected to Western blotting with the mouse anti-rotavirus capsid antibody, showing ~20-kDa $VP8^*$ protein band. The truncated $VP8^*$ protein band was also observed by Western blotting using the rabbit polyclonal antibody serum made against the truncated $VP8^*$ protein. This study suggested that the synthetic gene could be used as an easy way to produce the antigenic vaccine candidate for control of virus-associated diseases or to develop antibodies for diagnostic purpose.

Biological Analysis of Enzymatic Extracts from Capsosiphon Fulvescens Using the Microbulbifer sp. AJ-3 Marine Bacterium (해양미생물 Microbulbifer sp. AJ-3을 이용한 매생이 효소분해산물의 생리활성 연구)

  • Bae, Jeong-Mi;Cho, Eun-Kyung;Kim, Hye-Youn;Kang, Su-Hee;Choi, Young-Ju
    • Journal of Life Science
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    • v.22 no.5
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    • pp.627-633
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    • 2012
  • $Microbulbifer$ sp. AJ-3 was used to acquire the degrading products from $Capsosiphon$ $fulvescens$ (DPCF), which were investigated to determine its physiological activities. A crude enzyme extract from $Microbulbifer$ sp. AJ-3 hydrolyzes polysaccharide substrates such as agar, agarose, alginic acid, fucoidan, laminaran, starch, and chitin. Among them, agarose, laminaran, and alginic acid showed higher activities, especially alginic acid. The antioxidant activity of DPCF was measured by using 2,2-diphenyl-1-picryl-hydrazyl (DPPH) and superoxide dismutase (SOD)-like activities and were about 32% and 93% at 2 mg/ml, respectively. In addition, the nitrite-scavenging activity of DPCF was about 82%, 53%, and 12% at pH levels of 1.2, 3.0, and 6.0, respectively. To determine the influence of DPCF on alcohol metabolism, the generating activity of reduced-nicotinamide adenine dinucleotide (NADH) by alcohol dehydrogenase (ADH) was measured. The facilitating rate of ADH activity by DPCF was 130% at 2 mg/ml. The tyrosinase inhibitory activity of DPCF was slightly increased in a dose-dependent manner and was about 28% at 2 mg/ml. These results indicated that the enzymatic products from DPCF have a strong antioxidant, nitrite scavenging, and alcohol metabolizing activity.