Hidrolisis Protein Isolat Biji Melinjo (Gnetum gnemon L.) menggunakan Alkalase Terimobilisasi dan Aktivitasnya  sebagai Antihipertensi (Hydrolysis of Melinjo Seed (Gnetum gnemon L.) Isolated-Protein using Immobilized Alcalase and Its Activity as Antihyper

Hypertension is ranked third on the cause of death in Indonesia for all ages (6.8 %). Protein isolate of melinjo seeds (Gnetum gnemon L.) hydrolyzed with free alcalase has been known to have antihypertensive activity. However, the use of free alcalase is uneconomical since it can only be used once, thus we studied the immobilized alcalase effectivity to hydrolise the melinjo seeds protein isolate to be used as angiotensin converting enzyme (ACE) inhibitor. The immobilization method used entrapment method with DMDMOS/TMOS as matrix. Characterization of the immobilized enzyme was observed by FTIR and effectiveness of repeated hydrolysis was observed by the value of the degree of hydrolysis. The success of the hydrolysis process was determined by SDS-PAGE electrophoresis. Antihypertensive activity test was determined by the ability to inhibit ACE. The results showed that the alcalase has been immobilized in the matrix of silane and SDS-PAGE protein profile showed melinjo seed proteins had been successfully hydrolyzed. Hydrolysis process repeatedly demonstrated that immobilized alcalase was effective to hydrolyze protein isolates of melinjo seeds twice. Based on ACE inhibitory test, there were no significant differences between the protein isolate hydrolyzed by immobilized alcalase (Gg-PH), protein isolates hydrolyzed with free alcalase (Gg-PHB), and captopril. These findings suggest that the antihypertensive activity in Gg-PH is the same with that of captopril. Keywords: hypertension, antihypertensive, melinjo, enzymes immobilization, ACE inhibitor

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Revealing Antioxidant and Antidiabetic Potency of Melinjo (Gnetum Gnemon) Seed Protein Hydrolysate at Different Stages of Seed Maturation

Current Research in Nutrition and Food Science Journal ◽

10.12944/crnfsj.7.2.17 ◽

2019 ◽

Vol 7 (2) ◽

pp. 479-487 ◽

Cited By ~ 2

Author(s):

Anang Supriyadi ◽

Laras Sekar Arum ◽

Ari Satia Nugraha ◽

Anak Agung Istri Ratnadewi ◽

Tri Agus Siswoyo

Keyword(s):

Seed Protein ◽

Protein Hydrolysate ◽

Protein Profile ◽

Radical Scavenging ◽

Free Radical Scavenging ◽

Antidiabetic Activity ◽

Degree Of Hydrolysis ◽

Protein Extract ◽

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Gnetum Gnemon

roteins hydrolyzed from melinjo seeds (Gnetum gnemon) at green (GM), yellow (YM) and red (RM) stages of maturity were studied for their effectiveness in antioxidant and antidiabetic activities. The seed protein extract was hydrolyzed using alcalase 2.4L, and the resulting hydrolysates with the highest degree of hydrolysis, protein profile, and the most potent contributors to antioxidant and invitro antidiabetic activities were identified. The degree of hydrolysis value of hydrolysates ranged from 52-84%, and the SDS-PAGE protein profile showed two distinct bands in which the band with molecular weight of 30 kDa degraded more intensively. Antioxidant capacity was measured using different standard methods, including radical cation 2,2-azinobis-(3-ethylbenzothizoline-6-sulphonate) (ABTS•+) assay, hydroxyl radical (OH•), and superoxide anion (O2•-) scavenging. The green hydrolysate (GMH) had significantly higher (p<0.05) free radical scavenging (ABTS•+, OH•, and O2•-) activities than that of the yellow hydrolysate (YMH) and red hydrolysate (RMH). However, invitro antidiabetic testing was performed based on the inhibitory activity of α-amylase and α-glucosidase. GMH was found to be more effective than YMH and RMH. These results showed that the antioxidant and antidiabetic activity in hydrolyzed GM protein has high potential to be utilized as natural nutraceuticals.

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Functional properties of protein isolates from camelina (Camelina sativa (L.) Crantz) and flixweed (sophia, Descurainis sophia L.) seed meals

Food Production, Processing and Nutrition ◽

10.1186/s43014-021-00076-8 ◽

2021 ◽

Vol 3 (1) ◽

Author(s):

Na Thi Ty Ngo ◽

Fereidoon Shahidi

Keyword(s):

Functional Properties ◽

Protein Extraction ◽

Protein Profile ◽

Protein Solubility ◽

Protein Isolate ◽

Absorption Capacity ◽

Foaming Properties ◽

Oil Absorption ◽

Protein Isolates ◽

Water Holding

AbstractCamelina and flixweed (sophia) seed protein isolates were prepared using both the conventional extraction and ultrasonic-assisted extraction methods at 40 kHz for 20 min, and their functional properties investigated. SDS-PAGE showed that both ultrasound-assisted and conventional extractions resulted in a similar protein profile of the extract. The application of ultrasound significantly improved protein extraction/content and functional properties (water holding capacity, oil absorption capacity, emulsifying foaming properties, and protein solubility) of camelina protein isolate and sophia protein isolate. The water-holding and oil absorption capacities of sophia protein isolate were markedly higher than those of camelina protein isolate. These results suggest that camelina protein isolate and sophia protein isolate may serve as natural functional ingredients in the food industry. Graphical Abstract

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Enhanced Solubility of Rapeseed Meal Protein Isolates Prepared by Sequential Isoelectric Precipitation

Foods ◽

10.3390/foods9060703 ◽

2020 ◽

Vol 9 (6) ◽

pp. 703

Author(s):

Hristo Kalaydzhiev ◽

Radoslav Georgiev ◽

Petya Ivanova ◽

Magdalena Stoyanova ◽

Cristina L. M. Silva ◽

...

Keyword(s):

Protein Solubility ◽

Rapeseed Meal ◽

Protein Isolate ◽

Protein Fractions ◽

Isoelectric Precipitation ◽

Protein Isolates ◽

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Enhanced Solubility ◽

Meal Protein ◽

Biochemical Analyses

The solubility of plant protein isolates is a key determinant of their potential application. Two protein isolates (PI) from ethanol-treated industrial rapeseed meal, PI10.5–2.5 and PI2.5–8.5, were prepared by sequential isoelectric precipitation of alkali-extracted proteins (pH 12) starting from pH 10.5 to 2.5 or from pH 2.5 to 8.5, respectively. Biochemical analyses revealed that PI2.5–8.5 contained a higher amount of crude protein (72.84%) than PI10.5–2.5 (68.67%). In the same protein isolate, the level of total phenols (0.71%) was almost two-fold higher than that in PI10.5–2.5 (0.42%). No glucosinolates were established in both protein isolates. SDS-PAGE analysis demonstrated that PI10.5–2.5 contained 10 to 15 kDa protein fractions in a relatively higher amount, while PI2.5–8.5 was enriched in 18 to 29 kDa protein fractions. PI10.5–2.5 exhibited high solubility, varying from 41.74% at pH 4.5 to 65.13% at pH 6.5, while PI2.5–8.5 was almost two-fold less soluble under the same conditions. Up to pH 5.5, the addition of NaCl at 0.03 and 0.25 M diminished the solubility of PI2.5–8.5, while the solubility of PI10.5–2.5 was increased. The supplementation of PI10.5–2.5 with 0.25 M NaCl enhanced the protein solubility to 56.11% at pH 4.5 and 94.26% at pH 6.5. The addition of 0.03 M NaCl also increased the solubility of this protein isolate but to a lower extent. Overall, the approach for sequential precipitation of proteins influenced the biochemical characteristics, protein fractional profile and solubility of prepared protein isolates.

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Hydrolyzation of duck meat protein using Bacillus cereus TD5B protease, pepsin, trypsin and their potency as an angiotensin converting enzyme inhibitor

Journal of the Indonesian Tropical Animal Agriculture ◽

10.14710/jitaa.44.3.266-276 ◽

2019 ◽

Vol 44 (3) ◽

pp. 266

Author(s):

A. Winarti ◽

F. Rahmawati ◽

N. A. Fitriyanto ◽

J. Jamhari ◽

Y. Erwanto

Keyword(s):

Molecular Weight ◽

Bacillus Cereus ◽

Protein Concentration ◽

Ace Inhibitor ◽

Inhibiting Activity ◽

Converting Enzyme ◽

Meat Protein ◽

Hydrolysis Process ◽

Microbial Protease ◽

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This study was aimed to explore their potency of protein-bioactive of native ducks-meat after enzymatic hydrolysis by Bacillus-cereus TD5B-protease, Pepsin, and Trypsin as an angiotensin-converting enzyme (ACE) inhibitor. The samples: ducks-meats from 10 months age of male Mojosari and Magelang-Duck. The experiments: individually hydrolysis of meat-protein using protease-enzyme (0.1 % w/w) from Bacillus-cereus TD5B, Pepsin, or Trypsin. The observed parameters: protein concentration, protein molecular weight, ACE-inhibitor activity, and IC-value (IC50). Data of protein concentration were statistically analyzed using T-Test, while data of SDS-PAGE and ACE-inhibiting activity were analyzed descriptively. The results showed that soluble protein concentration increased due to the hydrolysis process, from 0.826±0.108 mg/mL to 1.050±0.197 mg/mL (Microbial-protease), 2.122±0.141 mg/mL (pepsin), 1.641±0.071 mg/mL (trypsin) for Mojosari-duck and 0.642±0.038 mg/mL to 1.171±0.534 mg/mL(Microbial-protease), 2.100±0.376 mg/mL(pepsin), 1.725±0.092 mg/mL(trypsin) for Magelang-duck. The SDS-PAGE pattern showed that there was a decrease of molecular weight of duck-meats due to the hydrolysis process, from the range of 196.53-43.88 kDa to the range of 71.35-10.12 kDa. Duck-meat protein hydrolysate had ACE-inhibiting activity 71.7%(Mojosari-Microbial-Protease) IC50 54μg/mL, 57%(Mojosari-Pepsin) IC50 151μg/mL, 75.8%(Mojosari-Trypsin) IC50 51μg/mL and 52.8%(Magelang-Microbial-Protease) IC50 83μg/mL, 78,5%(Magelang-Pepsin) IC50 85μg/mL, 83.9%(Magelang-Trypsin) IC50 22μg/mL. In conclusion, hydrolysate of Magelang duck-meat used Trypsin had better potency as an ACE-inhibitor.

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KAJIAN PENGARUH VARIASI KONSENTRASI ASAM SITRAT TERHADAP KEKUATAN GEL PRODUK GELATIN KULIT AYAM BROILER DIKAITKAN DENGAN POLA PROTEINNYA

Jurnal Kimia ◽

10.24843/jchem.2016.v10.i01.p16 ◽

2016 ◽

Cited By ~ 1

Author(s):

Tutut Hardikawati ◽

Ni Made Puspawati ◽

Ketut Ratnayani

Keyword(s):

Citric Acid ◽

Protein Profile ◽

Gel Strength ◽

Hydrolysis Process ◽

Collagen Protein ◽

Sds Page ◽

Chicken Skin ◽

Thermal Extraction ◽

Hydrolysis Of ◽

Collagen Tissue

Gelatin is a biopolymer that can be generated from partially hydrolysis of collagen tissue. Extraction of gelatin consists of pretreatment and thermal extraction steps. Pretreatment process used sodium hydroxide to remove non collagen protein in matrix sample, sulfuric acid to demineralize, and citric acid to hydrolyse. The aim of this research was to study the effect of variation in concentrations of citric acid used in hydrolysis process on the gel strength and protein profile of gelatin products extracted from broiler chicken skin. The variation of the concentration citric acid used was 0.7 % (GA); 1.5% (GB); and 3.0% b/v (GC) respectively. The gel strength was measured using CT3 Texture Analyzer and protein profile of gelatin product was analyzed by SDS-PAGE method. The result showed that variation in concentration of citric acid used in the pretreatment process affected the gel strength and protein profile of gelatin product. Increasing the concentration of citric acid used in pretreatment process decreased the gel strength and molecular weight of gelatin product. Gel strength of each gelatin product was 265.81 g bloom for GA ; 196.05 g bloom for GB (1.5%), and 35.32 g bloom for GC (3.0 %) respectively. The electropherogram of both GA (0.7%) and GB (1.5%) revealed similar pattern of protein bands but the thickness of each bands was different. On the other hands, GC (3.0%) did not show any protein bands on the eletropherogram. The best gelatin product obtained in this experiment was found by using 0.7 % b/v citric acid (GA) in the pretreatment process. The gelatin product (GA) had characteristics as follows: yield 15.73%; moisture 7.30%; ash 0.51%; protein content 97.95%; fat content 0.62%; gel strength 265. 81 g bloom and thicker protein bands than others.

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The Identification of Main Proteins Fractions from Pea Protein Isolates

Bulletin of University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca Agriculture ◽

10.15835/buasvmcn-agr:3752 ◽

1970 ◽

Vol 66 (2) ◽

Author(s):

Raul IANCHICI ◽

Rodica SEGAL ◽

Simona CRISAN ◽

Monica ZDREMTAN

Keyword(s):

Amino Acids ◽

Molecular Weight ◽

Human Body ◽

Nutritional Value ◽

Essential Amino Acids ◽

Protein Isolate ◽

Protein Isolates ◽

Pea Protein ◽

Molecular Weights ◽

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Pea’s proteins represent a valuable source of edible proteins which are well tolerating by human body, and contain all essential amino acids. Mature pea’s seeds are very rich in proteins that can be extracted in order to be used to improve the nutritional value of other foods. Pea’s proteins contain several fractions of albumins and globulins. We have used electrophoresis to separate and identify these fractions, to establish the molecular weight of each fraction, and also the proportion between them. The electrophoresis was conducted following the SDS-PAGE protocol. Using pea protein isolate as sample, we have found a number of 12 protein fractions with molecular weights ranging between 12500 and 140000 Daltons. From these, six fractions are prevalent.

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442 PB 247 BIOCHEMICAL CHARACTERIZATION OF CULTIVATED OPUNTIA SPECIES

HortScience ◽

10.21273/hortsci.29.5.494e ◽

1994 ◽

Vol 29 (5) ◽

pp. 494e-494

Author(s):

J.O. Kuti ◽

C.M. Galloway

Keyword(s):

Biochemical Characterization ◽

Protein Profile ◽

Seed Proteins ◽

Spectrophotometric Analysis ◽

Protein Profiles ◽

Genetic Studies ◽

Banding Patterns ◽

Molecular Weights ◽

Sds Page

The use of protein profiles and isozyme banding patterns as genetic markers in cultivated Opuntia species was investigated using SDS-PAGE and spectrophotometric analysis of seeds and stem (cladode) tissues. Twenty morphologically different entries belonging to six Opuntia species were analyzed for total protein profile and three enzyme systems (superoxide dismustase [SOD], phosphoglucomutase [PGM] and UDPG ppase). Seed proteins, mostly low molecular weights, were 3-fold that of cladode proteins. Marked differences in protein molecular weight were found among the entries. PGM activity, found only in the cladode tissues, differred among the entries. No UDPG ppase activity was found in either seeds or cladode tissues. Within the entries surveyed, identical SOD banding patterns were observed indicating some degree of similarity among the species. The preliminary results suggest that isozyme and protein profiles can be used as markers in genetic studies of cultivated Opuntia species.

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Comparative Analysis of Electrophoretic Profile of Major Proteins of Milk from Alpine and Carpathian Goats

Bulletin of University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca Veterinary Medicine ◽

10.15835/buasvmcn-vm:12447 ◽

2017 ◽

Vol 74 (1) ◽

pp. 20 ◽

Cited By ~ 1

Author(s):

Alina NASALEAN ◽

Laurentiu OGNEAN ◽

Sergiu MUNTEAN ◽

Stefana BALICI ◽

Horea MATEI

Keyword(s):

Protein Profile ◽

Milk Proteins ◽

Goat Milk ◽

Protein Composition ◽

Raw Milk ◽

Biologically Active ◽

Protein Fractions ◽

Animal Nutrition ◽

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Percentage Data

The milkâ€™s proteins provide nutritional and biologically active values, essential in human and animal nutrition. In the case of goat milk, the proteinsâ€™ concentration and quality represent basic indices for the evaluation of the nutritional and biologically active values. The proposal is to comparatively analyse the protein profile of milk. The milk was collected from two different breeds: French Alpine and Romanian Carpathian. During March and April 2016 there were collected samples of raw milk in hygienic and sanitation conditions. There were two lots: first lot has 10 Carpathian goats and the second lot has 10 Alpine goats. The protein composition of goat milk was established with SDS-PAGE, after the evaluation of the total proteinsâ€™ concentration with the Bradford method. The quantitative and percentage data obtained with electrophoresis revealed few differences between those 8 identified protein fractions. Between those two lots, regarding the levels of Î²-CN, k-CN and Î²-lactoglobulines there were significant differences. The other protein fractions have values almost identical. Statistical analysis of obtained data shaped the differences in the protein profile at those two breeds. Based on those differences it is to note the superior potential of the Alpine breed regarding the content in biologically active milk proteins. Regarding the obtained data, this study brings new contributions for the evaluation and analysis of protein profile as a nutritive and biologically active component of goat milk, confirming its character as a functional aliment.

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