• Title/Summary/Keyword: Enzyme Reaction

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Hydrolysis Methods for the Efficient Manufacture of Sugar Solutions from the Freshwater Alga Water-net (Hydrodictyon reticulatum) (담수조류 그물말로부터 당 용액의 효율적 제조를 위한 가수분해 방법)

  • Kim, Ji-Hyun;Kim, Sul Ki;Ko, Eun Hye;Kim, Jin-Cheol;Kim, Jin-Seog
    • Weed & Turfgrass Science
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    • v.2 no.2
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    • pp.176-183
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    • 2013
  • To explore hydrolysis methods for the efficient manufacture of sugar solutions from the freshwater alga Water-net (Hydrodictyon reticulatum, HR), acid hydrolysis, enzymatic hydrolysis, and combined hydrolysis (acid followed by enzymatic hydrolysis) were investigated. In the one-step acid hydrolysis, the reaction of 8% solids content using 2% sulfuric acid at $120^{\circ}C$ for 1 hour was desirable. In this case, glucose 27.44 g 100 g $DM^{-1}$ could be obtained from the HR-d13 samples. In the two-step acid hydrolysis, the primary hydrolysis (HR powder : 72% sulfuric acid = 1 g : 1.5 mL) was carried out for 1 hour at $60^{\circ}C$, and then the secondary hydrolysis was done for 1 hour at $120^{\circ}C$ after addition of distilled water 23.5 mL. In this case, glucose 35.11 g/100 g DM could be obtained from the HR-d13 samples. In the combined hydrolysis, 25% solids content using 2% hydrochloric acid were reacted for 1 hour at $120^{\circ}C$, and then citrate buffer and hydrolysis enzyme complexes (E1 1.0 mL+E2 0.2 mL $g^{-1}$ dried matter) were added and reacted for 1 - 2 days at $50^{\circ}C$. In this case, glucose 33.5 g 100 g $DM^{-1}$ could be obtained from the HR-d23+26 samples. In conclusion, combined hydrolysis was likely to be more useful saccharification method of HR biomass at a practical level, considering the glucose productivity, generation of fermentation-inhibiting substances (hydroxyl methyl furfural, furfural), and limited use of strong acid.

Efficient virus elimination for apple dwarfing rootstock M.9 and M.26 via thermotherapy, ribavirin and apical meristem culture (사과 왜성대목 M.9 및 M.26의 고온, ribavirin, 생장점 배양을 통한 바이러스 제거)

  • Kwon, Young Hee;Lee, Joung Kwan;Kim, Hee Kyu;Kim, Kyung Ok;Park, Jae Seong;Huh, Yoon Sun;Park, Eui Kwang;Yoon, Yeo Joong
    • Journal of Plant Biotechnology
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    • v.46 no.3
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    • pp.228-235
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    • 2019
  • Apple (Malus pumila) is one of the most economically important fruits in Korea. but virus infection has decreased the sustainable production of apples and caused serious problems such as yield loss and poor fruit quality. Virus or viroid infection including apple chlorotic leaf spot virus (ACLSV), apple stem pitting virus (ASPV), apple stem grooving virus (ASGV), apple mosaic virus (ApMV) and apple scar skin viroid (ASSVd) have been also reported in Korea. In many cases, as apple gets infected with virus and viroid with no specific symptoms, the damage and symptoms caused by the viruses are not detected. In our research, viruses in the rootstock were eliminated for a virus-free apple dwarfing rootstock of M.9 and M.26. The virus elimination methods were apical meristem culture, thermotherapy ($37^{\circ}C$, 6 weeks) and chemotherapy($Ribavirin^{(R)}$). The detection of apple viruses was accomplished by Enzyme-linked Immuno-Sorbent Assay (ELlSA) and reverse transcription-polymerase chain reaction (RT-PCR). RT- PCR method was 10 ~ 30% more sensitive than the ELISA method. The efficiency of virus elimination was enhanced in apical meristem culture method. The acquisition rate of virus-free apple dwarfing rootstocks was 30 ~ 40% higher in apical meristem culture. After the meristem culturing of M.9, the infection ratio of ACLSV, ASPV and ASGV was 45%, 60% and 50%, respectively. In the apple dwarfing rootstock of M.26, the infection ratio of ACLSV, ASPV and ASGV was 40%, 55% and 55%, respectively. Based on this study, the best method for the production of virus-free apple dwarfing rootstocks was the apical meristem culture.

Conditioned Media of RAW 264.7 Cells Stimulated with Phellinus linteus Extract Regulates the Epithelial-mesenchymal Transition in Prostate Cancer Cells (상황버섯에 의해 활성화된 RAW 264.7 대식세포주 배양액의 인간 전립선암 세포주의 epithelial-mesenchymal transition 조절)

  • Kang, Taewoo;An, Hyun-Hee;Park, Sul-Gi;Yu, Sun-Nyoung;Hwang, You-Lim;Kim, Ji-Won;Ahn, Soon-Cheol
    • Journal of Life Science
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    • v.29 no.8
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    • pp.904-915
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    • 2019
  • Prostate cancer (PCa) is one of the most metastatic tumor. Although hormone therapy or surgical castration is mostly conducted to treat PCa, it has a lot of side effects. Recently, many researchers have been exploring the tumor microenvironment to remedy these circumstances. Immune cells, especially macrophages, are an important composition of the tumor microenvironment. Under normal conditions, macrophages exhibit mild tumoricidal activity against tumors. However, once activated by interferon gamma or lipopolysaccharides, macrophages can kill cancer cells directly or indirectly by secreting cytokines and chemokines. In this study, murine macrophage RAW 264.7 cells were treated with Phellinus linteus extract. To analyze their pro-inflammatory phenotype, we were used several assays such as a real-time polymerase chain reaction, an enzyme-linked immunosorbent and nitric oxide assay. Prostate cancer cells were treated with the RAW 264.7-conditioned media, which was identified as a pro-inflammatory nature, for 48 h, and the expression of epithelial-mesenchymal transition (EMT)-related genes was determined. Not only N-cadherin, Snail, Twist, Slug, and Cadherin 11, which are mechenchymal-related proteins, were decrease, but epithelial marker of E-cadherin was increased. In addition, the mRNA level of vimentin, ccl2, and vegfa were decreased, as the EMT is closely related to the migration and invasion of cancer cells. In conclusion, the RAW 264.7-conditioned media stimulated with P. linteus extract inhibited migration and invasion and regulated the EMT pathway in human prostate cancer cells.

Anti-Hemolytic and Antimicrobial Effects against Multidrug-Resistant Bacteria of Enterococcus faecalis Isolated from Human Breast Milk (모유에서 분리한 Enterococcus faecalis의 다제내성 균에 대한 항용혈 및 항균 효과)

  • Yi, Eun-Ji;Lee, Jeong-eun;Jo, So-Yeon;Kim, Soo-bin;Yu, Du-na;Kook, Moochang;Kim, Ae Jung
    • Microbiology and Biotechnology Letters
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    • v.49 no.4
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    • pp.519-527
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    • 2021
  • In this study, the hemolysis of Enterococcus faecalis BMSE-HMP strains, isolated from human breast milk, was investigated, and the anti-hemolytic and antimicrobial effects on multidrug-resistant (MDR) bacteria were investigated. The enzyme activity of E. faecalis BMSE-HMP 4 strains was measured, and it was found that the activities of esterase and esterase lipase were the highest. In addition, no hemolytic reaction was observed in any of the isolates. Subsequently, the anti-hemolytic activity against MDR strains causing hemolysis was evaluated. E. faecalis BMSE-HMP002 had the highest anti-hemolytic activity against Staphylococcus aureus CCARM 3855 at 75.71 ± 10.00%. The anti-hemolytic activity against Escherichia coli DC 2 CCARM 0238 and Pseudomonas aeruginosa CCARM 0223 showed that the activity of BMSE-HMP001 was highest at 76.92 ± 2.99% and 87.93 ± 1.93%, respectively. Examination of the antimicrobial effects against the MDR bacteria Staphylococcus spp., Escherichia spp., Pseudomonas spp., Salmonella spp., Klebsiella spp., Enterobacter spp., and E. faecalis BMSE-HMP strains showed antimicrobial effects against both gram-positive and gram-negative strains. Breastfeeding delivers enterococci into the intestinal tract of newborns by lactation, and its usefulness is attracting attention as it has been reported that enterococci have a potential effect on neonatal immune development. In this study, the hemolytic and antimicrobial effects of E. faecalis BMSE-HMP strains on MDR bacteria were investigated, to confirm their potential as useful lactic acid bacteria. Additional studies on the antibiotic resistance and toxicity of the E. faecalis BMSE-HMP strains, isolated in this study, are necessary to prove it safe for use.

Inhibitory effect of water-soluble mulberry leaf extract on hepatic lipid accumulation in high-fat diet-fed rats via modulation of hepatic microRNA-221/222 expression and inflammation (고지방식이 급여 쥐에서 수용성 뽕나무 잎 추출물의 간 microRNA-221/222 발현 및 염증 조절을 통한 간 지질 축적억제 효과)

  • Lee, Mak-Soon;Kim, Cheamin;Ko, Hyunmi;Kim, Yangha
    • Journal of Nutrition and Health
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    • v.55 no.2
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    • pp.227-239
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    • 2022
  • Purpose: This study investigated the effects of water-soluble mulberry leaf extract (ME) on hepatic lipid accumulation in high-fat diet-fed rats via the regulation of hepatic microRNA (miR)-221/222 and inflammation. Methods: Male Sprague-Dawley rats (4 weeks old) were randomly divided into 3 groups (n = 7 each) and fed with 10 kcal% low-fat diet (LF), 45 kcal% high-fat diet (HF), or HF + 0.8% ME for 14 weeks. Lipid profiles and cytokine levels of the liver and serum were measured using commercial enzymatic colorimetric and enzyme-linked immunosorbent assay, respectively. The messenger RNA (mRNA) and miR levels in liver tissue were assayed by real-time quantitative reverse-transcription polymerase chain reaction. Results: Supplementation of ME reduces body weight and improves the liver and serum lipid profiles as compared to the HF group. The mRNA levels of hepatic peroxisome proliferator-activated receptor-gamma, sterol regulatory element binding protein-1c, fatty acid synthase, and fatty acid translocase, which are genes involved in lipid metabolism, were significantly downregulated in the ME group compared to the HF group. In contrast, the mRNA level of hepatic carnitine palmitoyl transferase-1 (involved in fatty acid oxidation) was upregulated by ME supplementation. Furthermore, administration of ME significantly downregulated the mRNA levels of inflammatory mediators such as hepatic tumor necrosis factor alpha (TNF-α), interleukin 6 (IL-6), monocyte chemoattractant protein-1, and inducible nitric oxide synthase. The serum levels of TNF-α, IL-6, and nitric oxide were also significantly reduced in ME group compared to the HF group. Expression of hepatic miR-221 and miR-222, which increase in the inflammatory state of the liver, were also significantly inhibited in the ME group compared to the HF group. Conclusion: These results indicate that ME has the potential to improve hepatic lipid accumulation in high-fat diet-fed rats via modulation of inflammatory mediators and hepatic miR-221/222 expressions.

Anti-inflammatory and Antioxidative Effects of Lotus Root Extract in LPS-PG-Stimulated Human Gingival Fibroblast-1 Cells (치주염 원인균 LPS-PG로 유도된 인체 치은섬유아세포에서 연뿌리 추출물에 대한 항염증 및 항산화 효과)

  • Lee, Young-Kyung;Kim, Chul Hwan;Jeong, Dae Won;Lee, Ki Won;Oh, Young Taek;Kim, Jeong Il;Jeong, Jin-Woo
    • Korean Journal of Plant Resources
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    • v.35 no.5
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    • pp.565-573
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    • 2022
  • Gingival inflammation is one of the main causes that can be related to various periodontal diseases. Human gingival fibroblast (HGF) is the major constituent in periodontal connective tissue and secretes various inflammatory mediators, such as nitric oxide (NO) and prostaglandin E2 (PGE2), upon lipopolysaccharide stimulation. This study is aimed at investigating the anti-inflammatory and antioxidative activities of Lotus Root extract (LRE) in Porphyromonas gingivalis derived lipopolysaccharide (LPS-PG)-stimulated HGF-1 cells. The concentration of NO and PGE2, as well as their responsible enzymes, inducible NO synthase (iNOS), and cyclooxygenase-2 (COX-2), was analyzed by Griess reaction, ELISA, and western blot analysis. LPS-PG sharply elevated the production and protein expression of inflammatory mediators, which were significantly attenuated by LRE treatment in a dose-dependent manner. LRE treatment also suppressed activation of Toll-like receptor 4 (TLR4)/myeloid differentiation primary response gene 88 (MyD88) and nuclear factor-κB (NF-κB) in LPS-PG-stimulated HGF-1 cells. In addition, one of phase II enzyme, NAD(P)H quinone dehydrogenase (NQO)-1, and its transcription factor, Nuclear factor erythroid 2-related factor 2 (Nrf2), were significantly induced by LRE treatment. Consequently, these results suggest that LRE ameliorates LPS-PG-induced inflammatory responses by attenuating TLR4/MyD88-mediated NF-κB, and activating NQO-1/Nrf2 antioxidant response element signaling pathways in HGF-1 cells.

Taste Compounds and Antioxidant Properties in Extracts of Angelica keiskei and Oenanthe javanica Juice By-Products According to Extraction Methods (추출 방법에 따른 명일엽과 돌미나리 착즙박의 정미성분 및 항산화 특성)

  • Hyun Jung Lee;Ha Na Ryoo;Hyeon Gyu Lee
    • Journal of Food Hygiene and Safety
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    • v.38 no.6
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    • pp.517-527
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    • 2023
  • This study aimed to examine the possibility of upcycling extracts of Angelica keiskei and Oenanthe javanica juice by-products through comparing enzyme extraction (EE) and complex extraction (CE) methods to increase the extraction yield and flavor of materials. A higher extraction yield was obtained for free amino acid content with EE and CE for A. keiskei and O. javanica juice by-products, respectively, and a higher extraction efficiency was achieved with juice by-products than with extracts prepared from raw materials before juice production. The content of major amino acids varied depending on the extraction method used. When used according to the characteristics of the extract, their use as a functional material was confirmed along with improvement in the flavor of the food. Consistently high extraction yields for organic acid and sugar levels were obtained with CE in A. keiskei and O. javanica juice by-products. The DPPH radical scavenging ability and TPC were consistently high with CE in A. keiskei and O. javanica juice by-products; the increase in extracted content was likely because of the reaction between the ethanol used for CE and the phenolic compounds. However, because the antioxidant capacity of the juice by-product extracts was somewhat lower than that of the extracts from raw materials before juice production, the amount used should be reviewed. The TFC was found to be higher in extracts obtained with EE than with CE for A. keiskei juice by-products; however, no significant difference was observed between EE and CE in the O. javanica juice by-products. Through this study, the taste compounds and antioxidant properties of extracts obtained from juice by-products produced after the production of A. keiskei and O. javanica green juice were analyzed, and the availability of high value-added materials was confirmed. Based on these research results, expanding specific R&D for practical use should be explored.