scholarly journals Variasi Profil Protein Daging Sapi yang Dibungkus Daun Pepaya dengan Elektrophoresis SDS-Page

2020 ◽  
Vol 22 (2) ◽  
pp. 46
Author(s):  
Tri Ade Saputro
Keyword(s):  
Medicinal Plant ◽  
Proteolytic Enzymes ◽  
Protein Molecule ◽  
Important Food ◽  
Food Ingredient ◽  
Digestive Process ◽  
Sds Page ◽  
Nutritional Needs

Abstrak Daging merupakan bahan pangan yang penting dalam memenuhi kebutuhan gizi.Protein pada daging lebih mudah dicerna dibanding protein yang berasal dari tumbuhan (nabati). Daun pepaya merupakan tanaman obat-obatan karena mengandung senyawa alkaloida dan enzim proteolitik, papain, khimopapain, dan lisosom,  yang  berguna pada proses pencernaan dan  mempermudah kerja usus Dalam pepaya terkandung enzim-enzim protease (pengurai protein) yaitu papain dan kimopapain. Kedua enzim ini mempunyai kemampuan menguraikan ikatan- ikatan dalam melekul protein sehingga protein terurai menjadi polipeptida dan dipeptide.Berdasarkan penelitian yang dilakukan di dapatkan hasil sebagai berikut pada tiap  sampelnya  sampel dengan rendaman  daun papaya selama 10  menit memiliki 11 pita protein, sampel rendaman daun papaya selama 20 menit memiliki 16 pita protein dan sampel rendaman daun papaya selama 30 menit memiliki 15 pita protein. Keywords: prifil protein, daging sapi, SDS page, elektroforesis AbstractBeef is an important food ingredient in meeting nutritional needs. Protein in beef is  easier  to  digest  than  protein  from  plants  (vegetables).  Papaya  leaf    is  a medicinal plant because it contains alkaloids and proteolytic enzymes, papain, khimopapain,  and  lyosomes,  which  are  useful  in  the  digestive  process  and facilitate the work of the intestines. Papaya contains protease enzymes (protein decomposers), papain and khimopapain. Both of these enzymes have the ability to decompose the bonds in the protein molecule so that the protein breaks down into polypeptides and dipeptides. Based on the research done in getting the following results in each sample, samples marinated with papaya leaf for 10 minutes have 11 protein bands, samples marinated during 20 minutes have 16 protein bands and samples marinated for 30 minutes have 15 protein bands. Kata Kunci: prifil protein, beef, SDS page, elektroforesis

scholarly journals Profil Protein Lima Jenis Daging yang Direndam Daun Pepaya Berbasis SDS-PAGE

2019 ◽  
Vol 2 (1) ◽  
pp. 132
Author(s):  
A.Meryam Susanti ◽  
Sri Darmawati ◽  
Endang Tri Wahyuni Maharani
Keyword(s):  
Broiler Chicken ◽  
Protein Profile ◽  
Major Protein ◽  
Protein Profiles ◽  
Important Food ◽  
Peptide Bonds ◽  
Free Range ◽  
Sds Page ◽  
Nutritional Needs ◽  
Minor Protein

Meat is an important food for fulfill nutritional needs, many of meats are consumed as a source of highest quality nutrition for humans, especially as a source of protein. Papaya leaves contain the enzyme papain (a protase enzyme that can hydrolyze proteins), so that it can be used to soften meat. The purpose of this study was to look at an overview of protein profiles in five types of meat, namely goat, beef, buffalo, free-range chicken and broiler chicken which were soaked in papaya leaves. The protein profile of five types of meat was analyzed using the SDS-PAGE 12% method. The results showed that the control meat of goat, beef, buffalo, free-range chicken and broiler chicken which were not soaked in papaya leaves showed that there were many major protein bands compared to minor protein bands. Whereas in goat, beef, buffalo, free-range chicken and broiler chicken which have been soaked in papaya leaves, the results were different compared to the control, there were many minor protein bands. While the major bands only have 6 to 9 protein bands. Based on these results indicate that immersion with the enzyme papain contained in papaya leaves can break down peptide bonds, if it works on meat it can be broken down so the meat becomes tender and protein bands in the form of micromolecules.Daging merupakan bahan pangan yang penting dalam memenuhi kebutuhan gizi, banyak dikomsumsi sebagai sumber nutrisi yang berkualitas bagi manusia terutama sebagai sumber protein. Daun pepaya mengandung enzim papain (enzim protase yang dapat menghidrolisa protein), sehingga dapat digunakan untuk melunakkan daging. Tujuan penelitian ini untuk melihat gambaran profil protein pada lima jenis daging yaitu daging kambing, sapi, kerbau, ayam kampung dan ayam potong yang direndam daun pepaya. Profil protein lima macam daging dianalisis menggunakan metode SDS-PAGE 12%. Hasil penelitian menunjukkan pada daging kontrol yaitu daging kambing, sapi, kerbau, ayam kampung dan ayam potong yang tidak direndam daun pepaya menunjukkan terdapat banyak pita protein mayor dibandingkan pita protein minor. Sedangkan pada daging kambing, sapi, kerbau, ayam kampung dan ayam potong yang telah direndam daun pepaya menunjukkan hasil yang berbeda dibandingkan dengan kontrol yaitu pada semua daging terdapat banyak pita protein minor. Sedangkan pita mayor hanya terdapat 6 sampai 9 pita protein saja. Berdasarkan hasil tersebut menunjukkan bahwa perendaman dengan enzim papain yang terdapat dalam daun pepaya dapat memecah ikatan peptida, jika bekerja pada daging dapat diuraikan sehingga daging menjadi empuk, dan pita protein berbentuk mikromolekul.

scholarly journals BACTERIAL AGGLUTINATION BY A LECTIN FROM THE LEAVES OF THE MEDICINAL PLANT, PIMENTA DIOICA (L.) MERR

2018 ◽  
Vol 10 (4) ◽  
pp. 35 ◽  
Author(s):  
Surya P H ◽  
Elyas K K ◽  
Deepti Madayi
Keyword(s):  
Escherichia Coli ◽  
Medicinal Plant ◽  
Biochemical Characterization ◽  
Exchange Chromatography ◽  
Bacterial Typing ◽  
Gram Negative ◽  
Sds Page ◽  
Bacterial Filters ◽  
Pimenta Dioica ◽  
Bacterial Agglutination

Objective: The current investigation involves the purification, characterization of the lectin from the leaves of Pimenta dioica (L.) Merr. (Myrtaceae) a medicinal plant, and its application in bacterial typing.Methods: A lectin was purified from the leaves by cation exchange chromatography. SDS PAGE revealed the molecular weight of the purified lectin. Biochemical characterization was carried out by performing various tests. Hemagglutination inhibition was conducted to detect the sugar specificity. Additionally, bacterial agglutination was performed to predict whether the purified lectin was able to agglutinate the bacterial strains.Results: SDS PAGE analysis revealed the lectin to be a tetramer in the range of 43-66 kDa. The purified lectin agglutinated human, avian, and mouse erythrocytes, and was inhibited by 125 mmol of mannose and xylose. The lectin was stable at 0-60 ° C for 30 min and was unaffected by either 2-Mercaptoethanol (2-ME) or Dithiothreitol (DTT) (50-250µM). A pH of 6.0–8.0 was found optimum for its activity and was nearly independent of metal ions. The purified lectin contained about 20% carbohydrate as estimated by Anthrone method. Purified lectin agglutinated the Gram-negative Escherichia coli and Proteus vulgaris.Conclusion: The isolated lectin was found to possess significant hemagglutinating activity. Due to its ability to agglutinate Gram negative bacteria such as Escherichia coli and Proteus vulgaris, it could be used for bacterial typing and for the design of bacterial filters.

scholarly journals Implementasi Forward Chaining dan Metode Simpleks untuk Menentukan Ransum Unggas yang Ekonomis

2020 ◽  
Vol 6 (1) ◽  
Author(s):  
Wildan Mahmud
Keyword(s):  
Simplex Method ◽  
Poultry Feed ◽  
Nutritional Content ◽  
Food Ingredient ◽  
Food Ingredients ◽  
Forward Chaining ◽  
Nutritional Needs ◽  
Nutritional Element ◽  
Selection Of

Poultry need food to maintain their lives and to produce. To meet the needs of these nutritional elements, poultry eat from various types of food. Because there is no single food ingredient that contains a complete nutritional element, so we need a variety of appropriate and balanced food ingredients to meet nutritional needs. This can be done by choosing cheap food ingredients without ignoring the quality of the nutritional content that is adjusted to the type and age of poultry. To solve the above problems, an expert system with a forward chaining method can be used to determine the nutritional needs and nutritional content of food ingredients, while linear programming with the simplex method is used to determine the combination of poultry feed-forming ingredients that meet economical poultry nutritional needs. The combination of the forward chaining method and the simplex method can accelerate the preparation of combinations of food ingredients, so that the resulting application can provide a solution in making decisions for the selection of food ingredients forming ration effectively

scholarly journals CONSTRUCTION, EXPRESSION AND PURIFICATION OF RECOMBINANT PRE-MATURE PEPTIDE OF PLANTARICIN F FROM Lactobacillus plantarum S34 IN Escherichia coli

2015 ◽  
Vol 16 (1) ◽  
pp. 31
Author(s):  
Kusdianawati Kusdianawati ◽  
Apon Zaenal Mustopa ◽  
Suharsono Suharsono ◽  
Bugi Ratno Budiarto ◽  
Fatimah Fatimah ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Lactobacillus Plantarum ◽  
Proteolytic Enzymes ◽  
Leader Peptide ◽  
Human Consumption ◽  
Mature Peptide ◽  
Expression And Purification ◽  
Food Preservative ◽  
E Coli ◽  
Sds Page

Plantaricin is one of bacteriocins that have the potential to be used as food preservative. Plantaricin is safe for human consumption because it can be easily degraded by proteolytic enzymes. The objective of this study was to express and purify recombinant pre-mature peptide of plantaricin F from <em>Lactobacillus plantarum</em> S34 in <em>Escherichia coli</em>. Plantaricin gene-specific primer was used to obtain pln F structural gene amplicon from L. <em>plantarum</em> S34. This amplicon was cloned in pET32a vector and expressed in E. coli BL21 (DE3) pLysS. Pre-mature plantaricin F peptide was expressed as Histagged-fusion protein and separated by Co2+-chelating affinity chromatography. L. <em>plantarum</em> S34-derived pre-mature plantaricin F peptide fused with thioredoxin-(His)6tag had successfully been expressed in E. <em>coli</em> BL21 (DE3) pLysS using pET32a as an expression vector. The fused recombinant pln F as pre-mature state expressed had a molecular mass of +24 kDa, meanwhile the fused recombinant that contained only the leader peptide of pln F appeared as +20 kDa based on SDS-PAGE separations. The optimal production of fused recombinant pln F as soluble fraction was obtained when culture condition was added with 0.5 mM of IPTG and incubated at 22°C for 5 hours (OD~1). Furthermore, the expression of fused recombinant pln F as its pre-mature peptide pointed out that the pln F’s leader peptide could be proteolytically cleaved by a system in heterologous cells. Overall, heterologous pln F production as pre-mature peptide fused with thioredoxin-(His)6tag had been well established. From this research, we expect plantaricin F can be expressed and purified in E. coli.

scholarly journals The Reactivity and Allergenic Potential of Hazelnut Peptides

2013 ◽  
Vol 70 (1) ◽  
pp. 25
Author(s):  
Lavinia Florina Calinoiu ◽  
Dan Cristian Vodnar ◽  
Carmen Socaciu
Keyword(s):  
Proteolytic Enzymes ◽  
Peptide Fragments ◽  
Dot Blot ◽  
Specific Antibodies ◽  
Allergenic Potential ◽  
Sds Page ◽  
Allergenic Proteins ◽  
Life Threatening ◽  
Specific Proteins ◽  
Sensitive Method

The aim of this paper was to focus on proteins present in some food products, like hazelnuts and to investigate their allergenic potential. Several techniques were used to characterize these extracted proteins, with respect to their composition, degradability by digestive proteolytic enzymes and their reactivity with specific antibodies. It was important to analyse which proteins were present in the hazelnuts, to see if there were proteins present to trigger an allergic reaction and if the digestion enzymes trypsin and pepsin influence the presence of the (allergic) protein compounds. Allergies to tree nuts and seeds can cause life-threatening and sometimes fatal reactions. To examine the properties of Hazelnut protein it was important to solubilize it by extraction. After extraction, it was investigated how hazelnut protein can be modified by proteases and what the effect was on the immune reaction. The Bradford method is a fast and sensitive method to determine the concentration of soluble protein. When the Bradford reagent (Coomassie Brilliant Blue) binds to the protein, the colour changes from red to purple and the absorption maximum changes from 495 to 595 nm. The value obtained as the final concentration of proteins was 7.3495. SDS-PAGE is a method to separate mixtures of proteins by electrophoresis. Protein molecules are negatively charged by binding of SDS molecules; subsequently they are separated in an electric field. Their differences in size (molecular weight) leads to separation. In this case the method is used to follow proteolytic degradation of hazelnut proteins (allergens) by intestinal proteases (trypsin, pepsin). A different, more specific and sensitive method is immunoblotting (Western Blot) in which the SDS-PAGE separated proteins are transferred from the gel to a membrane and specific antibodies are used in a series of reactions to visualize specific allergens on this membrane. The remarked spots represented a positive identification of allergenic proteins. This means that peptide fragments of various size, produced during the digestion of a protein can still be immunological active. As it was shown there was still reactivity between proteins and specific antibodies. The Dot Blot is a simple immunoblotting technique used to detected specific proteins in a mixture of different proteins and/or other molecules. No separation technique prior to the actual immuno-detection is necessary. Also, Dot Blot confirmed the presence of allergenic proteins made visible through the light spots on the membrane.

Bacteriocin ST91KM, produced by Streptococcus gallolyticus subsp. macedonicus ST91KM, is a narrow-spectrum peptide active against bacteria associated with mastitis in dairy cattle

2008 ◽  
Vol 54 (7) ◽  
pp. 525-531 ◽  
Author(s):  
Reneé Pieterse ◽  
Svetoslav D. Todorov ◽  
Leon M.T. Dicks
Keyword(s):  
Staphylococcus Aureus ◽  
Antimicrobial Activity ◽  
Streptococcus Agalactiae ◽  
Staphylococcus Epidermidis ◽  
Mode Of Action ◽  
Proteolytic Enzymes ◽  
Streptococcus Dysgalactiae ◽  
Streptococcus Gallolyticus ◽  
Streptococcus Uberis ◽  
Sds Page

Streptococcus gallolyticus subsp. macedonicus ST91KM produces a bacteriocin (macedocin ST91KM) active against Streptococcus agalactiae , Streptococcus dysgalactiae subsp. dysgalactiae , Streptococcus uberis , Staphylococcus aureus , and Staphylococcus epidermidis . Macedocin ST91KM is, according to tricine-SDS PAGE, between 2.0 and 2.5 kDa in size. Antimicrobial activity remained unchanged after 2 h of incubation at pH 2.0–10.0 and after 100 min at 100 °C. The peptide was inactivated after 20 min at 121 °C and when treated with proteolytic enzymes. Treatment with α-amylase had no effect on activity, suggesting that the mode of action does not depend on glycosylation. Amplification of the genome of strain ST91KM with primers designed from the macedocin precursor gene (mcdA) produced 2 fragments (approximately 375 and 220 bp) instead of one 150-bp fragment, as recorded for macedocin produced by Streptococcus gallolyticus subsp. macedonicus ACA-DC 198. Strain ACA-DC 198 was not available. However, DNA amplified from strain LMG 18488 (ACA-DC 206), genetically closely related to strain ACA-DC 198, revealed 99% homology to the mcdA of strain ACA-DC 198 (accession No. DQ835394). Macedocin ST91KM may thus be a second putative bacteriocin described for Streptococcus gallolyticus subsp. macedonicus.

scholarly journals Effects of Different Softening Processes on the Hardness and Quality of Thawed Neritic Squid (Uroteuthis edulis) Muscle

Processes ◽  
2020 ◽  
Vol 8 (2) ◽  
pp. 135
Author(s):  
Mark J. Grygier ◽  
Yu-Wen Fan ◽  
Wen-Chieh Sung
Keyword(s):  
Proteolytic Enzymes ◽  
Senior Citizens ◽  
Enzyme Treatment ◽  
Hot Air Drying ◽  
Air Drying ◽  
Hot Air ◽  
Sds Page ◽  
Chewing Ability ◽  
Orbital Shaking

While attempting to develop a soft, seafood-based product as a potential food item for senior citizens, we evaluated the effects of different softening methods on the hardness and quality of thawed muscular mantle tissue of the neritic squid (Uroteuthis edulis) versus controls. Comparisons were made among injection with proteolytic enzymes (papain, bromelain); soaking in these enzymes or in alkali solutions (NaHCO3, NaOH); various soaking regimes combined with either orbital shaking under vacuum, ultrasonic processing, or ultrasonic cleaning; or hot-air drying and rehydration. Elderly panelists’ sensory impressions of thawed and heat-sterilized squid mantle subjected to these treatments were recorded, together with the total volatile basic nitrogen (TVBN), pH, color, protein breakdown profile (SDS-PAGE), and histological characteristics of thawed squid mantle subjected to the same treatments but not heat-sterilized. TVBN measurements showed that squid mantle remained in a close-to-fresh state under all treatments except for hot-air drying. The pH and hardness decreased and the muscles turned white when soaked in either enzymes or alkali. Orbital shaking under vacuum while soaking in 0.3% papain for 12 h produced the softest product, the next softest being obtained by injection with 0.3% papain. After orbital shaking under vacuum in 1.0% papain, protein degradation was confirmed by SDS-PAGE, and broken muscle fibers were evident in histological sections. Sensory evaluation panelists with unimpeded chewing ability rated mushy, papain-treated squid mantle poorly. Soaking in 2% NaHCO3 in an ultrasonic processor, following by washing out of the alkali, proved to be a better tenderizing method than either enzyme treatment or hot-air drying for neritic squid mantle intended for consumption by senior citizens.

Novel bacteriocins produced by Geobacillus stearothermophilus

2009 ◽  
Vol 4 (2) ◽  
pp. 196-203 ◽  
Author(s):  
Karina Pokusaeva ◽  
Nomeda Kuisiene ◽  
Dziuginta Jasinskyte ◽  
Kazimiera Rutiene ◽  
Jordana Saleikiene ◽  
...  
Keyword(s):  
Antimicrobial Agents ◽  
Direct Detection ◽  
Proteolytic Enzymes ◽  
Geobacillus Stearothermophilus ◽  
Inhibitory Peptides ◽  
Heat Stable ◽  
Sds Page ◽  
Ph Range ◽  
Agriculture And Food ◽  
Logarithmic Growth

AbstractFour novel heat-stable bacteriocin-like substances were found to be produced by Geobacillus stearothermophilus strains isolated from oil-wells in Lithuania. Geobacillus stearothermophilus 32A, 17, 30 and 31 strains were identified as producers of bacteriocins with bactericidal activity against closely related Geobacillus species and several pathogenic strains: Bacillus cereus DSM 12001 and Staphylococcus haemolyticus P903. The secretion of the analysed bacteriocins started during early logarithmic growth and dropped sharply after the culture entered the stationary phase of growth. The antimicrobial activity of the bacteriocins against sensitive indicator cells disappeared after treatment with proteolytic enzymes, indicating their proteinaceous nature. Bacteriocins were stable throughout the pH range between 4 and 10, and no loss in activity was noted following temperature exposures up to 100°C. Direct detection of antibacterial activity on SDS-PAGE suggests that the inhibitory peptides have a molecular weight of 6–7.5 kDa. Such bacteriocins with broad activity spectra, including antipathogenic action, are attractive to the biotechnology industry as they could be used as antimicrobial agents in medicine, agriculture and food products.

scholarly journals Purification and Characterization of an Extracellular Protease from Penicillium chrysogenum Pg222 Active against Meat Proteins

2002 ◽  
Vol 68 (7) ◽  
pp. 3532-3536 ◽  
Author(s):  
María J. Benito ◽  
Mar Rodríguez ◽  
Félix Núñez ◽  
Miguel A. Asensio ◽  
María E. Bermúdez ◽  
...  
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Penicillium Chrysogenum ◽  
Proteolytic Enzymes ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Maximum Activity ◽  
Extracellular Protease ◽  
Collagenolytic Activity ◽  
Sds Page

ABSTRACT An extracellular protease from Penicillium chrysogenum (Pg222) isolated from dry-cured ham has been purified. The purification procedure involved several steps: ammonium sulfate precipitation, ion-exchange chromatography, filtration, and separation by high-performance liquid chromatography. Based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis and gel filtration, the purified fraction showed a molecular mass of about 35 kDa. The hydrolytic properties of the purified enzyme (EPg222) on extracted pork myofibrillar proteins under several conditions were evaluated by SDS-PAGE. EPg222 showed activity in the range of 10 to 60°C in temperature, 0 to 3 M NaCl, and pH 5 to 7, with maximum activity at pH 6, 45°C, and 0.25 M NaCl. Under these conditions the enzyme was most active against tropomyosin, actin, and myosin. EPg222 showed collagenolytic activity but did not hydrolyze myoglobin. EPg222 showed higher activity than other proteolytic enzymes like papain, trypsin, and Aspergillus oryzae protease. The N-terminal amino acid sequence was determined and was found to be Glu-Asn-Pro-Leu-Gln-Pro-Asn-Ala-Pro-Ser-Trp. This partial amino acid sequence revealed a 55% homology with serine proteases from Penicillium citrinum. The activity of this novel protease may be of interest in ripening and generating the flavor of dry-cured meat products.

Chitosan Metal Complex: An Artificial Metalloprotease Hydrolyzing Trypsin Inhibitor from Soybean

2012 ◽  
Vol 430-432 ◽  
pp. 414-418
Author(s):  
Peng Liang ◽  
Dong Feng Wang ◽  
Xing Yu Wang ◽  
Mei Ding ◽  
Li Zhang ◽  
...  
Keyword(s):  
Reaction Time ◽  
Metal Complex ◽  
Metal Complexes ◽  
Trypsin Inhibitor ◽  
Rate Constants ◽  
Proteolytic Enzymes ◽  
Protein Cleavage ◽  
Reaction Conditions ◽  
Sds Page ◽  
Optimum Reaction

Most popular agents for protein cleavage are proteolytic enzymes, but they require more rigorous hydrolytic conditions, and it is very difficulty to separate them from the productions. In this paper, an new artificial metalloprotease, Cu(II) complex of cyclen (Cu(II)Cyc) using chitosan as supporter, was designed and synthesized successfully. The hydrolytic efficient under different conditions was measured and confirmed with HPLC, SDS-PAGE, further more. Some reaction conditions, such as pH, temperature, were researched, and the results show that the optimum reaction time was 48 h, temperature was 60 °C respectively, The observed rate constants for trypsase inhibitor cleavage was 1.006×10-2 h-1 under above conditions, and the observed rate constants was 2.120×10-2 h-1 when the pH was 9.0. This paper suggested that the chitosan metal complexes could efficiently accelerate the hydrolysis reaction.

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