ISOLATION of a AMYLASE INHIBITORS from MUNGBEAN and SOYBEAN and INHIBITORY EFFECT on HUMAN SALIVARY and PORCINE PANCREATIC AMYLASE

2013 ◽  
Vol 14 (1) ◽  
Author(s):  
Budiasih Wahyuntari ◽  
Martinus Nicoadi Tekol Tekol
Keyword(s):  
Inhibitory Effect ◽  
Pancreatic Amylase ◽  
Sds Page ◽  
Amylase Inhibitors

Penghambat alfa amilase adalah salah satu komponen dalam suplemen makanan yang telah lama digunakan untuk terapi kegemukan karena penghambat amilase mempengaruhi metabolisme karbohidrat dalam sistem pencernaan. Sejumlah peneliti melaporkan terdapat dua grup penghambat amilase, yaitu protein dan non protein. Penghambat protein dilaporkan terdapat dalam kelompok kacang-kacangan dan biji-bijian. Tujuan penelitian ini adalah untuk mengisolasi penghambat protein yang terdapat dalam kacang hijau dan kedele. Kacang hijau dan kedele merupakan kacang-kacangan yang penting dalam makanan popular di Indonesia. Penghambat protein diendapkan dengan konsentrasi bertingkat garam ammonium sulfat [(NH)4SO4] dari 30-70%. Penghambat protein diuji terhadap amilase saliva manusia (HSA) danamilase pankreas babi (PPA), serta kestabilannya terhadap pemanasan pada 100oC selama 30 menit. Hasil penelitian menunjukkan bahwa semua endapan jenuh dari semua konsentrasi amonium sulfat yang diuji menghambat PPA, tetapi tidak semua endapan jenuh tersebut dapatmenghambat HSA. Hanya semua endapan jenuh (NH)4SO4 dari kacang hijau yang dapat menghambat HSA, dan penghambatan tertinggi terhadap HSA adalah endapan jenuh (NH)4SO4 50%. Endapan jenuh (NH)4SO4 40 % dari kedele putih dan endapan jenuh (NH)4SO4 60% dari kedele hitam dengan masing-masing penghambatan 98.67; 26.86 and27.63%. Endapan jenuh (NH)4SO4 60-70% dari kacang hijau, 50% dari kedele putih dan 50% dari kedele hitam menghambat PPA 100%. Pemanasan penghambat pada 100oC selama 30 menit hampir tidak mempengaruhi penghambatannya terhadap PPA. Profil protein juga diamati menggunakan analisis SDS/PAGE.

Purification and Characterization of Lupus Anticoagulant Like Protein from Agkistrodon Halys Brevicaudus Venom

1996 ◽  
Vol 76 (06) ◽  
pp. 0993-0997
Author(s):  
Zhao-Yan Li ◽  
Xiao-Wei Wu ◽  
Tie-Fu Yu ◽  
Eric C-Y Lian
Keyword(s):  
Amino Acid ◽  
Lupus Anticoagulant ◽  
Inhibitory Effect ◽  
Clotting Factor ◽  
Acid Oxidase ◽  
Crude Venom ◽  
Amino Acid Oxidase ◽  
Sds Page ◽  
The Individual ◽  
Reducing Condition

SummaryBy means of CM-Sephadex C-25, DEAE-Sephadex A-50, Sephadex G-200, and Sephadex G-75 chromatographies, a lupus anticoagulant like protein (LALP) from Agkistrodon halys brevicaudus was purified. On SDS-PAGE, the purified LALP had a molecular weight of 25,500 daltons under non-reducing condition and 15,000 daltons under reducing condition. The isoelectric point was pH 5.6. Its N terminal amino acid sequencing revealed a mixture of 2 sequences: DCP(P/S)(D/G)WSSYEGH(C/R)Q(Q/K). It was devoid of phospho-lipaseA, fibrino(geno)lytic, 5′-nucleotidase, L-amino acid oxidase, phosphomonoesterase, phosphodiesterase and thrombin-like activities, which were found in crude venom. In the presence of LALP, PT, aPTT, and dRVVT of human plasma were markedly prolonged and its effects were concentration-dependent but time-independent. The inhibitory effect of LALP on the plasma clotting time was enhanced by decreasing phospholipid concentration in TTI test. The individual clotting factor activity was not affected by LALP when higher dilutions of LALP-plasma mixture were used for assay. Russell’s viper venom time was shortened when high phospholipid confirmatory reagent was used. Therefore, the protein has lupus anticoagulant property.

Structure-based design of human pancreatic amylase inhibitors from the natural anthocyanin database for type 2 diabetes

2020 ◽  
Vol 11 (4) ◽  
pp. 2910-2923 ◽  
Author(s):  
Lianghua Xie ◽  
Jianling Mo ◽  
Jingdan Ni ◽  
Yang Xu ◽  
Hongming Su ◽  
...  
Keyword(s):  
Type 2 Diabetes ◽  
Design Approach ◽  
Pancreatic Amylase ◽  
Amylase Inhibitor ◽  
Amylase Inhibitors

Malvidin 3-O-arabinoside is identified as a novel human pancreatic amylase inhibitor from the natural anthocyanin database with a structure-based design approach.

Inhibitory Effect of Hexane Fraction from Myagropsis myagroides on Pancreatic ��-Amylase In Vitro

2015 ◽  
Vol 25 (3) ◽  
pp. 328-333 ◽  
Author(s):  
Won-Min Pak ◽  
Koth-Bong-Woo Ri Kim ◽  
Min-Ji Kim ◽  
Ji-Young Cho ◽  
Dong-Hyun Ahn
Keyword(s):  
Inhibitory Effect ◽  
Hexane Fraction ◽  
Pancreatic Amylase ◽  
Myagropsis Myagroides

scholarly journals Impact of B-Ring Substitution and Acylation with Hydroxy Cinnamic Acids on the Inhibition of Porcine α-Amylase by Anthocyanin-3-Glycosides

Foods ◽  
2020 ◽  
Vol 9 (3) ◽  
pp. 367 ◽  
Author(s):  
Julia A. H. Kaeswurm ◽  
Lisa Könighofer ◽  
Melanie Hogg ◽  
Andreas Scharinger ◽  
Maria Buchweitz
Keyword(s):  
Inhibitory Effect ◽  
Postprandial Blood Glucose ◽  
Titration Calorimetry ◽  
Plant Metabolites ◽  
Diabetes Mellitus Type Ii ◽  
Secondary Plant Metabolites ◽  
Vis Spectroscopy ◽  
Postprandial Blood Glucose Level ◽  
The Impact ◽  
Amylase Inhibitors

An inhibitory effect on α-amylase and α-glucosidase is postulated for polyphenols. Thus, ingestion of those secondary plant metabolites might reduce postprandial blood glucose level (hyperglycemia), which is a major risk factor for diabetes mellitus type II. In addition to a previous study investigating structure−effect relationships of different phenolic structures, the effect of anthocyanins is studied in detail here, by applying an α-amylase activity assay, on the basis of the conversion of 2-chloro-4-nitrophenyl-4-O-ß-galactopyranosyl maltoside (GalG2CNP) and detection of CNP release by UV/Vis spectroscopy and isothermal titration calorimetry (ITC). All anthocyanin-3-glucosides showed a mixed inhibition with a strong competitive proportion, Kic < 134 µM and Kiu < 270 µM; however, the impact of the B-ring substitution was not statistically significant. UV/Vis detection failed to examine the inhibitory effect of acylated cyanidins isolated from black carrot (Daucus carota ssp. Sativus var. Autrorubens Alef.). However, ITC measurements reveal a much stronger inhibitory effect compared to the cyanidin-3-glucoside. Our results support the hypothesis that anthocyanins are efficient α-amylase inhibitors and an additional acylation with a cinnamic acid boosts the observed effect. Therefore, an increased consumption of vegetables containing acylated anthocyanin derivatives might help to prevent hyperglycemia.

A Novel Dysfibrinogenemia with Abnormal γ-Chain(Fibrinogen Nagoya).

1979 ◽  
Author(s):  
Junki Takamatsu ◽  
Kanji Oqata ◽  
Tadashi Kamiya ◽  
Katsuo Koie ◽  
Takagi Takashi ◽  
...  
Keyword(s):  
Amino Acid ◽  
Inhibitory Effect ◽  
Chain Structure ◽  
Japanese Family ◽  
Electrophoretic Mobilities ◽  
Sds Page ◽  
History Of ◽  
The Difference ◽  
Fibrin Monomers ◽  
Major Defect

Six individuals in 3 generations of Japanese family had prolonged thrombin clotting time, but no history of hemorrhagic or thrombotic disease. Very low fibrinogen levels were obtained by thrombin clottable protein, while immunological procedures gave normal values of fibrinogen. The serum contained 40-80μg/ml of unclottable fibrinogen related antigens. The patients’ plasma had an inhibitory effect on the fibrin formation in normal plasma. Major defect of this fibrinogen was a delayed aggregation of fibrin monomers.On CM-chromatography (CM-52) of the S-carboxymethylated fibrinogen, three different γ-chains, named γx, γL and γR, were separated. They did not differ in their electrophoretic mobilities in SDS-PAGE, but were distinguishable in PAGE containing 8M urea. Moreover, the amino acid compositions and tryptic peptide mappings of each chain revealed a little difference from those of normal fibrinogen γ chains, suggesting the difference in amino acid substitution or oligosaccharide chain structure.Based on these findings, we designated this fibrinogen as fibrinogen Nagoya; its possible identity without other dysfibrinogenemia has not been excluded.

scholarly journals Purification and characterization of pig kidney aminopeptidase P. A glycosyl-phosphatidylinositol-anchored ectoenzyme

1990 ◽  
Vol 267 (2) ◽  
pp. 509-515 ◽  
Author(s):  
N M Hooper ◽  
J Hryszko ◽  
A J Turner
Keyword(s):  
Chelating Agents ◽  
Inhibitory Effect ◽  
Purification And Characterization ◽  
Glycosyl Phosphatidylinositol ◽  
Pig Kidney ◽  
Sds Page ◽  
Cyclic Phosphate ◽  
Aminopeptidase P ◽  
Mild Acid

Aminopeptidase P (EC 3.4.11.9) was solubilized from pig kidney membranes with bacterial phosphatidylinositol-specific phospholipase C (PI-PLC) and then purified by a combination of anion-exchange and hydrophobic-interaction chromatographies. Contaminating peptidase activities were removed by selective affinity chromatography. The purified enzyme was apparently homogeneous on SDS/PAGE with an Mr of 91,000. Enzymic deglycosylation revealed that aminopeptidase P is a glycoprotein, with up to 25% by weight of the protein being due to the presence of N-linked sugars. The phospholipase-solubilized aminopeptidase P was recognized by an antiserum to the cross-reacting determinant (CRD) characteristic of the glycosyl-phosphatidylinositol anchor. This recognition was abolished by mild acid treatment or deamination with HNO2, indicating that the CRD was due exclusively to the inositol 1,2-cyclic phosphate ring epitope generated by the action of PI-PLC. The activity of aminopeptidase P was inhibited by chelating agents and was stimulated by Mn2+ or Co2+ ions, confirming the metallo-enzyme nature of this peptidase. Selective inhibitors of other aminopeptidases (actinonin, amastatin, bestatin and puromycin) had little or no inhibitory effect.

scholarly journals cAMP suppresses p21ras and Raf-1 responses but not the Erk-1 response to granulocyte-colony-stimulating factor: possible Raf-1-independent activation of Erk-1

1997 ◽  
Vol 322 (1) ◽  
pp. 79-87 ◽  
Author(s):  
Xavier F. CSAR ◽  
Alister C. WARD ◽  
Brigitte W. HOFFMANN ◽  
Graeme G. GUY ◽  
John A. HAMILTON
Keyword(s):  
Inhibitory Effect ◽  
Intracellular Camp ◽  
Two Dimensional ◽  
Granulocyte Colony Stimulating Factor ◽  
Colony Stimulating Factor ◽  
Camp Analogue ◽  
Extracellular Signal ◽  
Sds Page ◽  
Stimulating Factor

The cAMP analogue 8-bromo-cAMP (8BrcAMP) inhibits granulocyte-colony-stimulating factor (G-CSF)-stimulated DNA synthesis in myeloid NFS-60 cells. We examined the effect of 8BrcAMP addition on the G-CSF-stimulated extracellular signal-related protein kinase 1 (Erk-1), p21ras and Raf-1 activation. The Erk-1 activity was not down-regulated by the increase in intracellular cAMP levels, whereas p21ras and Raf-1 activities were, suggesting that Erk-1 activity might not be dependent on upstream p21ras and/or Raf-1 activity in this system. To explore this possibility further, we sought to determine whether there were downstream substrates of Raf-1 that were distinguishable from those of Erk-1 by using two-dimensional SDS/PAGE analysis of the protein phosphorylation patterns of NFS-60 cell cytosolic extracts treated with exogenous Raf-1 or Erk-1 in the presence of [γ-32P]ATP. The two phosphorylation patterns were found to have many differences. To gain further insights into the possible relevance of these phosphorylation patterns and as an approach to exploring in more detail the inhibitory effect of 8BrcAMP, two-dimensional SDS/PAGE analysis was performed on the cytosolic extracts of 32P-labelled NFS-60 cells treated with G-CSF, in the absence or presence of 8BrcAMP. It was found that the phosphorylated proteins whose appearance was specific to the action of exogenous Raf-1 were sensitive to the action of 8BrcAMP in vivo, whereas those whose appearance was specific to the action of exogenous Erk-1 alone, or common to the actions of Raf-1 and Erk-1, were 8BrcAMP-insensitive. The results are consistent with a Raf-1-independent pathway for Erk-1 activation in G-CSF-treated myeloid cells, and a number of potential downstream substrates of these kinases have been identified for further characterization.

scholarly journals Protease Inhibitors Purified from the Canola Meal Extracts of Two Genetically Diverse Genotypes Exhibit Antidiabetic and Antihypertension Properties

Molecules ◽  
2021 ◽  
Vol 26 (7) ◽  
pp. 2078
Author(s):  
Saira Hussain ◽  
Ata-ur Rehman ◽  
David J. Luckett ◽  
S.M. Saqlan Naqvi ◽  
Christopher L. Blanchard
Keyword(s):  
Protease Inhibitors ◽  
Serine Proteases ◽  
Plant Pathogens ◽  
Gel Filtration ◽  
Ace Inhibition ◽  
Inhibitory Effect ◽  
Canola Meal ◽  
Native Page ◽  
Dpp Iv ◽  
Sds Page

Valorization of vegetable oil waste residues is gaining importance due to their high protein and polyphenol contents. Protease inhibitors (PIs), proteins from these abundantly available waste residues, have recently gained importance in treating chronic diseases. This research aimed to use canola meal of genetically diverse Brassica napus genotypes, BLN-3347 and Rivette, to identify PIs with diverse functionalities in therapeutic and pharmacological applications. The canola meal PI purification steps involved: native PAGE and trypsin inhibition activity, followed by ammonium sulfate fractionation, anion exchange, gel filtration, and reverse-phase chromatography. The purified PI preparations were characterized using SDS-PAGE, isoelectric focusing (IEF), and N terminal sequencing. SDS-PAGE analysis of PI preparations under native reducing and nonreducing conditions revealed three polymorphic PIs in each genotype. The corresponding IEF of the genotype BLN-3347, exhibited three acidic isoforms with isoelectric points (pI) of 4.6, 4.0, and 3.9, while Rivette possessed three isoforms, exhibiting two basic forms of pI 8.65 and 9.9, and one acidic of pI 6.55. Purified PI preparations from both the genotypes displayed dipeptidyl peptidase-IV (DPP-IV) and angiotensin-converting enzyme (ACE) inhibition activities; the BLN-3347 PI preparation exhibited a strong inhibitory effect with lower IC50 values (DPP-IV 37.42 µg/mL; ACE 129 µg/mL) than that from Rivette (DPP-IV 67.97 µg/mL; ACE 376.2 µg/mL). In addition to potential human therapy, these highly polymorphic PIs, which can inhibit damaging serine proteases secreted by canola plant pathogens, have the potential to be used by canola plant breeders to seek qualitative trait locus (QTLs) linked to genes conferring resistance to canola diseases.

scholarly journals The Effect of Bioactive Polyphenols from Anacardium occidentale Linn. Leaves on α-Amylase and Dipeptidyl Peptidase IV Activities

2020 ◽  
Vol 20 (5) ◽  
pp. 1010
Author(s):  
Nur Imanina Abdullah Thaidi ◽  
Hanapi Mat Jusoh ◽  
Ahmad Badruddin Ghazali ◽  
Deny Susanti ◽  
Normah Haron
Keyword(s):  
Diabetes Mellitus ◽  
Type 2 Diabetes ◽  
Dipeptidyl Peptidase ◽  
Inhibitory Effect ◽  
Dipeptidyl Peptidase Iv ◽  
Anacardium Occidentale ◽  
Mature Leaves ◽  
Young Leaves ◽  
Amylase Inhibitors

Anacardium occidentale Linn. (A. occidentale L.) leaves possess bioactive polyphenols which are associated with antidiabetic potency for the management of type 2 diabetes mellitus (T2DM). In this study, free, soluble ester, and insoluble-bound phenolic fractions from young and mature leaves of A. occidentale L. were extracted. Subsequently, all fractions were investigated for their inhibitory effect on α-amylase and dipeptidyl peptidase IV (DPPIV) activities. Both free (72.45 ± 3.6%) and soluble ester (83.40 ± 4.7%) phenolic fractions in the mature leaves extracts had significantly demonstrated greater α-amylase inhibitors than the young leaves. Likewise, soluble ester (4.09 ± 0.34 µg/mL) and insoluble-bound (4.87 ± 0.32 µg/mL) phenolic fractions in the mature leaves extracts were significantly more effective in inhibiting DPPIV than the young leaves. As for fractions comparison, insoluble-bound derived from the young leaves extract was a more potent α-amylase inhibitor than free and soluble ester phenolic fractions (p < 0.0001). Besides, soluble ester and insoluble-bound phenolic fractions showed a stronger inhibitor of DPPIV than the free phenolic (p < 0.001), irrespective of the maturity of the leaves. In conclusion, this study showed that A. occidentale L. extracts possessed antidiabetic properties, which may potentially be used as an alternative treatment for T2DM management.

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