• Title/Summary/Keyword: α-Amylase

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New Action Pattern of a Maltose-forming α-Amylase from Streptomyces sp. and its Possible Application in Bakery

  • Ammar, Youssef Ben;Matsubara, Takayoshi;Ito, Kazuo;Iizuka, Masaru;Limpaseni, Tipaporn;Pongsawasdi, Piamsook;Minamiura, Noshi
    • BMB Reports
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    • v.35 no.6
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    • pp.568-575
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    • 2002
  • An $\alpha$-amylase (EC 3.2.1.1) was purified that catalyses the production of a high level of maltose from starch without the attendant production of glucose. The enzyme was produced extracellularly by thermophilic Streptomyces sp. that was isolated from Thailand's soil. Purification was achieved by alcohol precipiation, DEAE-Cellulose, and Gel filtration chromatographies. The purified enzyme exhibited maximum activity at pH 6-7 and $60^{\circ}C$. It had a relative molecular mass of 45 kDa, as determined by SDS-PAGE. The hydrolysis products from starch had $\alpha$-anomeric forms, as determined by $^1H$-NMR. This maltose-forming $\alpha$-amylase completely hydrolyzed the soluble starch to produce a high level of maltose, representing up to 90%. It hydrolyzed maltotetrose and maltotriose to primarily produce maltose (82% and 62%, repectively) without the attendant production of glucose. The high maltose level as a final end-product from starch and maltooligosaccharides, and the unique action pattern of this enzyme, indicate an unusual maltose-forming system. After the addition of the enzyme in the bread-baking process, the bread's volume increased and kept its softness longer than when the bread had no enzyme.

The non-saponin fraction of Korean Red Ginseng (KGC05P0) decreases glucose uptake and transport in vitro and modulates glucose production via down-regulation of the PI3K/AKT pathway in vivo

  • Park, Soo-Jeung;Lee, Dasom;Kim, Dakyung;Lee, Minhee;In, Gyo;Han, Sung-Tai;Kim, Sung Won;Lee, Mi-Hyang;Kim, Ok-Kyung;Lee, Jeongmin
    • Journal of Ginseng Research
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    • v.44 no.2
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    • pp.362-372
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    • 2020
  • Background: The non-saponin fraction of Korean Red Ginseng has been reported to have many biological activities. However, the effect of this fraction on anti-diabetic activity has not been elucidated in detail. In this study, we investigated the effects of KGC05P0, a non-saponin fraction of Korean Red Ginseng, on anti-diabetic activity in vitro and in vivo. Methods: We measured the inhibition of commercially obtained α-glucosidase and α-amylase activities in vitro and measured the glucose uptake and transport rate in Caco-2 cells. C57BL/6J mice and C57BLKS/Jdb/db (diabetic) mice were fed diets with or without KGC05P0 for eight weeks. To perform the experiments, the groups were divided as follows: normal control (C57BL/6J mice), db/db control (C57BLKS/Jdb/db mice), positive control (inulin 400 mg/kg b.w.), low (KGC05P0 100 mg/kg b.w.), medium (KGC05P0 200 mg/kg b.w.), and high (KGC05P0 400 mg/kg b.w.). Results: KGC05P0 inhibited α-glucosidase and α-amylase activities in vitro, and decreased glucose uptake and transport rate in Caco-2 cells. In addition, KGC05P0 regulated fasting glucose level, glucose tolerance, insulin, HbA1c, carbonyl contents, and proinflammatory cytokines in blood from diabetic mice and significantly reduced urinary glucose excretion levels. Moreover, we found that KGC05P0 regulated glucose production by down-regulation of the PI3K/AKT pathway, which inhibited gluconeogenesis. Conclusion: Our study thereby demonstrated that KGC05P0 exerted anti-diabetic effects through inhibition of glucose absorption and the PI3K/AKT pathway in in vitro and in vivo models of diabetes. Our results suggest that KGC05P0 could be developed as a complementary food to help prevent T2DM and its complications.

Investigation of Antioxidant, Hypoglycemic and Anti-Obesity Effects of Euphorbia Resinifera L.

  • Benrahou, Kaoutar;El Guourrami, Otman;Mrabti, Hanae Naceiri;Cherrah, Yahia;My El Abbes, Faouzi
    • Journal of Pharmacopuncture
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    • v.25 no.3
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    • pp.242-249
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    • 2022
  • Objectives: The aim of this work is to evaluate the in vitro antioxidant, hypoglycemic, and antiobesity effects of Euphorbia resinifera extracts and investigate the phenolic constituents and the toxicity of these extracts. Methods: Phytochemical screening was performed to detect polyphenols and flavonoids. Antioxidant activity was evaluated by four methods (DPPH, ABTS, H2O2, and xanthine oxidase inhibition). The hypoglycemic effect was determined by the inhibition of α-amylase and α-glucosidase enzymes in vitro and via a starch tolerance study in normal rats. The antiobesity effect was estimated by in vitro inhibition of lipase. Results: Phytochemical screening revealed that the ethanolic extract was rich in polyphenols (99 ± 0.56 mg GEA/g extract) and tannins (55.22 ± 0.17 mg RE/g extract). Moreover, this extract showed higher antioxidant activity in different tests: the DPPH assay (IC50 = 53.81 ± 1.83 ㎍/mL), ABTS assay (111.4 ± 2.64 mg TE/g extract), H2O2 (IC50 = 98.15 ± 0.68 ㎍/mL), and xanthine oxidase (IC50 = 10.26 ± 0.6 ㎍/mL). With respect to hypoglycemic effect, the aqueous and ethanolic extracts showed IC50 values of 119.7 ± 2.15 ㎍/mL and 102 ± 3.63 ㎍/mL for α-amylase and 121.4 ± 1.88 and 56.6 ± 1.12 ㎍/mL for α-glucosidase, respectively, and the extracts lowered blood glucose levels in normal starch-loaded rats. Additionally, lipase inhibition was observed with aqueous (IC50 = 25.3 ± 1.53 ㎍/mL) and ethanolic (IC50 = 13.7 ± 3.03 ㎍/mL) extracts. Conclusion: These findings show the antioxidant, hypoglycemic, and hyperlipidemic effects of E. resinifera extracts, which should be investigated further to validate their medicinal uses and their pharmaceutical applications.

Analysis of Chlorogenic Acid Content and Biological Activities of Aralia elata Ethanol Extract (두릅 에탄올 추출물의 Chlorogenic acid 함량 분석 및 생리활성)

  • Lee, Jeong Ho;Jeong, Kyoung Ok;Im, So Yeon;Jin, Da Mon;Lee, Wang Ro
    • Korean Journal of Plant Resources
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    • v.35 no.5
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    • pp.574-585
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    • 2022
  • This study was conducted to quantify chlorogenic acid content and evaluate biological activity, such as antioxidant, antibacterial, anti-inflammatory, and digestive enzyme activity of Aralia elata ethanol extract (AEE). The SC50 of DPPH and ABTS radical scavenging activities of AEE were 4.79±0.05 mg/mL, 5.79±0.05 mg/mL; total polyphenol and total flavonoid contents were 170.0±1.8 mgGAE/g, 105.5±4.1 mgQE/g, respectively. Nitric oxide (NO) was increased in RAW 264.7 cells and Caco-2 cells with treatment of LPS, and production of NO was inhibited by AEE in a concentration-dependent manner. Production of NO was reduced by 60.0±1.1% in RAW 264.7 cells and 50.7±2.8% in Caco-2 cells at of AEE. Similarly, the production of inflammatory cytokines (TNF-α, IL-1β and IL-6) was inhibited in a concentration dependent manner. Antibacterial activity increased as the dose concentration of AEE increased, and the MIC was 75 mg/mL for L. monocytogenes, and 100 mg/mL for S. typhimurium and H. pylori. In addition, amylase and protease enzyme activity was observed in AEE and increased enzyme activity was observed according to the concentration of the extract. AEE contained 7.06±0.01 mg/g of chlorogenic acid. As a result of the experiment, it is judged that it can be used as basic data for the development of health food using Aralia elata.

Physiological Characteristics and Anti-Diabetic Effect of Pediococcus pentosaceus KI62

  • Kim, Seulki;Hong, Sang-pil;Lim, Sang-Dong
    • Food Science of Animal Resources
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    • v.41 no.2
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    • pp.274-287
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    • 2021
  • The purpose of this study is to examine the physiological characteristics and anti-diabetic effects of Pediococcus pentosaceus KI62. The α-amylase and α-glucosidase inhibitory activity of P. pentosaceus KI62 was 94.86±3.30% and 98.59±0.52%, respectively. In MRS broth containing 3% maltodextrin inoculated by P. pentosaceus KI62, the amounts of short chain fatty acids (SCFA) were propionic acid 18.05±1.85 mg/kg, acetic acid 1.12±0.07 g/100 mL, and butyric acid 2.19±0.061 g/kg, and those of medium chain fatty acids (MCFA) were C8 0.262±0.031 mg/kg, C10 0.279±0.021 mg/kg, and C12 0.203±0.009 mg/kg. Compared to sixteen antibiotics, P. pentosaceus KI62 had the highest sensitivity to penicillin-G and rifampicin, as well as the highest resistance to vancomycin and ampicillin. The strain also showed higher leucine arylamidase and valine arylamidase activities than other enzyme activities, but it did not produce β-glucuronidase which is carcinogenic enzymes. The survival rate of P. pentosaceus KI62 in 0.3% bile was 91.67%. Moreover, the strain showed a 98.63% survival rate in pH 2.0. P. pentosaceus KI62 exhibits resistance to Escherichia coli, Salmonella Typhimurium, Listeria monocytogenes, and Staphylococcus aureus at rates of 29.41%, 38.10%, 51.72%, and 50.47%, respectively. P. pentosaceus (23.31%) showed a similar adhesion ability to L. rhamnosus GG, the positive control (24.49%). These results show that P. pentosaceus KI62 has possibility as a probiotic with anti-diabetic effects.

Silymarin attenuates escitalopram (cipralex) induced pancreatic injury in adult male albino rats: a biochemical, histological, and immunohistochemical approach

  • Rasha Mamdouh Salama;Sara Gamal Tayel
    • Anatomy and Cell Biology
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    • v.56 no.1
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    • pp.122-136
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    • 2023
  • Depression is a prevalent global problem since ages, predominately treated with SSRI. Cipralex, is an antidepressant of the SSRIs class used as a remedy for mood, depression and anxiety. Silymarin (SIL), a natural free radical scavenging, has an antioxidant and anti-inflammatory properties. This hypothesis evaluates, for the first time, the role of cipralex on the structure of the endocrine and exocrine components of the pancreas and assess the beneficial effects of SIL on these changes. Forty-five rats were divided into control, cipralex, and cipralex plus SIL groups. During sacrifice, all rats and pancreases were weighed and the ratio of pancreatic weight (PW) to rat weight (RW) was calculated, blood samples were collected to estimate fasting glucose, insulin and amylase levels, the specimens were prepared for histological, immunohistochemical (inducible nitric oxide synthase [iNOS], tumour necrosis factor-alpha [TNF-α], caspase 3, proliferating cell nuclear antigen [PCNA], and anti-insulin antibody), and morphometrical studies. Cipralex group exhibited marked destruction of the pancreatic architecture of the exocrine and endocrine parts, with a dense collagen fiber deposition. Also, there is highly significant decrease (P<0.001) of PW/RT ratio, insulin, and amylase levels, the number and diameter of islets of Langerhans, the number of PCNA positive immunoreactive cells, and the number of insulin positive β-cells. Furthermore, a highly significant increase of glucose level, iNOS, TNF-α, and caspase-3 positive immunoreactive cells in the islets of Langerhans and acinar cells were observed. SIL improves the pancreatic histological architecture, weight loss, biochemical, and immunohistochemical analyses. Administering SIL is advantageous in managing cipralex induced pancreatic injury via its anti-inflammatory, antioxidant, and anti-apoptotic qualities.

Effect of Oral Administration of Intact Casein on Gastrointestinal Hormone Secretion and Pancreatic α-Amylase Activity in Korean Native Steer

  • Lee, S.B.;Choi, C.W.;Jin, Y.C.;Wang, T.;Lee, K.H.;Ku, M.B.;Hwang, J.H.;Kim, K.H.;Vega, R.S.A.;Lee, H.G.
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.5
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    • pp.654-660
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    • 2013
  • Three Korean native steers ($779{\pm}24$ kg) fitted with duodenal cannulas were used in a $3{\times}3$ Latin square design to investigate the influence of oral administration of soluble proteins, intact casein (IC) and acid hydrolyzed casein (AHC), on gastro-intestinal hormone (GIH) secretion in the blood and pancreatic ${\alpha}$-amylase activity in the duodenum. Oral treatment consisted of a basic diet (control), IC (C+100% protein), or AHC (C+80% amino acid, 20% peptide) for 21 d. Blood and duodenum samples were collected for measurement of serum GI hormones, and pancreatic ${\alpha}$-amylase activity was determined at 900, 1030, 1330, 1630, and 1930 h after feeding on d 21 of treatment. The levels of serum cholecystokinin (CCK) and secretin in the IC treatment group were higher compared to the other treatment groups (p<0.05). In addition to the changes in CCK and secretin levels upon IC treatment, the pancreatic ${\alpha}$-amylase activity in the duodenum was higher in the IC group compared to the control diet group (p<0.05). The response of serum ghrelin to IC and AHC treatment was in accordance with the response of serum secretin. The level of peptide fragments flowing in the duodenum was higher in the IC treatment group than the other treatment groups (p<0.05). In conclusion, this study demonstrated that an increase in duodenal CCK and secretin upon IC oral administration increased pancreatic ${\alpha}$-amylase secretion. In addition, ghrelin may be associated with GI hormone secretion in Korean native steers.

Multifunctional Probiotic and Functional Properties of Lactiplantibacillus plantarum LRCC5314, Isolated from Kimchi

  • Yoon, Seokmin;Cho, Hyeokjun;Nam, Yohan;Park, Miri;Lim, Ahyoung;Kim, Jong-Hwa;Park, Jaewoong;Kim, Wonyong
    • Journal of Microbiology and Biotechnology
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    • v.32 no.1
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    • pp.72-80
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    • 2022
  • In this study, the survival capacity (acid and bile salt tolerance, and adhesion to gut epithelial cells) and probiotic properties (enzyme activity-inhibition and anti-inflammatory activities, inhibition of adipogenesis, and stress hormone level reduction) of Lactiplantibacillus plantarum LRCC5314, isolated from kimchi (Korean traditional fermented cabbage), were investigated. LRCC5314 exhibited very stable survival at ph 2.0 and in 0.2% bile acid with 89.9% adhesion to Caco-2 intestinal epithelial cells after treatment for 2 h. LRCC5314 also inhibited the activities of α-amylase and α-glucosidase, which are involved in elevating postprandial blood glucose levels, by approximately 72.9% and 51.2%, respectively. Treatment of lipopolysaccharide (LPS)-stimulated RAW 264.7 cells with the LRCC5314 lysate decreased the levels of the inflammatory factors nitric oxide, tumor necrosis factor (TNF-α), interleukin (IL)-1β, and interferon-γ by 88.5%, 49.3%, 97.2%, and 99.8%, respectively, relative to those of the cells treated with LPS alone. LRCC5314 also inhibited adipogenesis in differentiating preadipocytes (3T3-L1 cells), showing a 14.7% decrease in lipid droplet levels and a 74.0% decrease in triglyceride levels, as well as distinct reductions in the mRNA expression levels of adiponectin, FAS, PPAR/γ, C/EBPα, TNF-α, and IL-6. Moreover, LRCC5314 reduced the level of cortisol, a hormone with important effect on stress, by approximately 35.6% in H295R cells. L. plantarum LRCC5314 is identified as a new probiotic with excellent in vitro multifunctional properties. Subsequent in vivo studies may further demonstrate its potential as a functional food or pharmabiotic.

Immunostimulatory activity and structural characteristics of neutral polysaccharides isolated from ginseng leaves fermented by Cordyceps sinensis (동충하초균으로 발효한 인삼잎에서 분리한 중성다당의 면역활성 및 구조적 특성)

  • Cha, Ha Young;Park, Hye-Ryung;Shin, Kwang-Soon
    • Korean Journal of Food Science and Technology
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    • v.53 no.5
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    • pp.570-577
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    • 2021
  • This study elucidated the biological activities and structural characteristics of polysaccharides isolated from ginseng leaves fermented using Cordyceps sinensis (GLF). GLF comprised at least 18 glycosyl linkages, including 4-linked glucose residues (24.0%). To characterize the neutral polysaccharides in GLF, it was further fractionated by anion exchange chromatography, and the unabsorbed fraction (GLF1) was isolated. Peritoneal macrophages stimulated with GLF1 produced various cytokines in a dose-dependent manner. The properties and activities of the four subfractions (PHI, PHIA1-PHIA3) obtained after sequential enzymatic digestion were examined. PHI and PHIA3 primarily comprised glucose, whereas PHI exhibited an iodine-color reaction. Furthermore, the PHIA1-3 fractions indicated that cytokine production was completely inhibited. These results suggest at the immune activities of GLF1 may be due to the α-(1→4)-glucan branched at the C(O)6 position, which was produced by C. sinensis.

Changes in Microorganisms, Enzyme Activities and Taste Components of Kochujang Added with Maesil Extract during Fermentation

  • Lee, Min-Ji;Seog, Eun-Ju;Lee, Jun-Ho
    • Preventive Nutrition and Food Science
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    • v.12 no.1
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    • pp.51-57
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    • 2007
  • The effects of maesil extract addition on the palatability and quality of kochujang, a traditional Korean condiment, were investigated in terms of the microbial characteristics, enzyme activities, and taste components during 100 days of fermentation. Viable cell counts of bacteria and yeast in maesil extract-added kochujang (maesil kochujang) were increased in proportion to added maesil extract from 1 to 5% during fermentation, up to 80 and 20 days, respectively and maintained $5.0{\sim}14.5{\times}10^6$ CFU/g. Activities of α-amylase, $\beta$-amylase, and protease were also increased in proportion to added maesil extract up to 20, 20, and 60 days, respectively and were higher than those of control during the aging process. The major organic acids in maesil kochujang were citric and malic acid. Composition and content of free sugar were not changed remarkably in general by the addition of maesil extract except maltose. The major free sugars of maesil kochujang were in the order of glucose>sucrose>maltose, and glucose content decreased significantly as the ratio of maesil extract increased, while maltose content increased significantly (p<0.05).