Premium Essay

Isolation and Characterization of Proteins

In:

Submitted By stevenvlfrt
Words 1129
Pages 5
Isolation and Characterization of Proteins: Hydrolysis and Chromatography
Niña Maria Inez Torno, Kimberly Victoriano, Steven John Villafuerte, Klarissa Viloria, Liana Yamzon
Group 8 2F-Pharmacy Biochemistry Laboratory

ABSTRACT
Chemical analysis can be either qualitative or quantitative in nature. In qualitative analysis, we work to identify the substances present in a given sample. We are only concerned whether certain substances are present or absent while in quantitative analysis, we are concerned in determining the amount of each component present in a sample. This experiment aims to isolate and characterize the intact protein in wheat flour which is Gluten through hydrolysis and chromatography. Gluten was isolated then hydrolyzed by proteolytic enzyme and was subjected to different qualitative tests. Paper Chromatography was also performed to analyze the different amino acid components of gluten. Tests showed that Gluten has several amino acid components.
INTRODUCTION

Proteins are a class of organic compounds which are present in and vital to every living cell. In the form of skin, hair, callus, cartilage, muscles, tendons and ligaments, proteins hold together, protect, and provide structure to the body of a multi-celled organism. In the form of enzymes, hormones, antibodies, and globulins, they catalyze, regulate, and protect the body chemistry. In the form of hemoglobin, myoglobin and various lipoproteins, they affect the transport of oxygen and other substances within an organism.
It is also known as polypeptides, are organic compounds made of amino acids arranged in a linear chain and folded into a globular form. The amino acids in a polymer are joined together by covalent bonds called peptide bonds between the carboxyl and amino groups of adjacent amino acid residues. The physical and chemical properties unique to each amino acid are the result

Similar Documents

Premium Essay

Isolation, Identification of Microbes Isolated from Thar Desert

...ISOLATION,IDENTIFICATION AND CHARACTERIZATION AND APPLICATIONS OF MICROBES ISOLATED FROM THAR DESERTSOIL,INDIA INTRODUCTION This study was done to identify the characterization, isolation and identification of microorganism from the desert soil. In the present study, thermal resistant and osmotolerant bacteria from the Thar soils were reported. Bacteria were isolated from 20 soil samples of 100gm each, collected from sand dunes. As Thar Desert were also evaluated for their plant growth promontory effects in terms of seed germination, shoot and root length elongation and total biomass production. It also tells about the occurrence of nodulated legumes. Most of the bacteria grow best in normal environmental conditions, but some grow in extreme conditions like extremes of pH, temperature, high salt reservoirs of microorganisms, as bacteria, protozoans, yeasts, molds, and microscopic worms are routinely found in this environment and are continuously submitted to environmental and anthropogenic perturbations which can lead to serious modifications of microbial taxonomic and functional diversity present in desert soil, therefore some of the fungi like Thermophilic fungi are also present in these conditions. Moreover DNA Based Identification and Characterization of microorganisms were also done from the soil of Thar Desert. Along with this, application of the microorganism isolated from the thar desert are like in biofertiliser, for medicinal parouses etc., example Ricinus communis...

Words: 800 - Pages: 4

Free Essay

A Comparitive Study on Invertase Production, Characterization & Optimization by Penicillium Brevicompactum & Penicillium Chrysogenum on Pineapple Peel Waste.

...INTRODUCTION Microorganisms are used extensively to provide a vast range of products and services. They have proved to be particularly useful because of the ease of their mass cultivation, speed of growth, use of cheap substrates (which in many cases are wastes) and the diversity of potential products. Their ability to readily undergo genetic manipulation has also opened up almost limitless further possibilities for new products and services from the fermentation industries (Trevor Palmor, 2004). Microorganisms are a large and disease group that exist as its single cell or cell cultures. These include bacteria, fungi, algae, protozoa and infectious agents at the borderline of life. Microbes are present most abundantly in soil, atmosphere and water plays a important role in the biochemical agent for the conversion of complex organic compounds into simple organic compounds (Glazer and Nikaido, 1995). Microorganisms are closely associated with health and welfare of human beings. Some Microorganisms are beneficial and others are detrimental. For example, Microorganisms are involved in the making of yoghurt, cheese and wine, in the production of Penicillin, interferons and alcohol, and the processing of industrial and domestic wastes. Microorganisms can cause disease, spoil food and deteriorate materials like iron, pipes, glass lenses and wood pilings (Glazer and Nikaido, 1995). Most of these Microorganisms irrespective of their origins, were subsequently modified by conventional...

Words: 11029 - Pages: 45

Premium Essay

Advantages Of Proteomics

...introduced in 1995. Proteomics is the characterization and identification of all proteins that expressed by a genome or tissue and understanding how these proteins function (Mohanty, 2005). Besides, the purpose of proteomics is not only to recognize all the proteins in a cell but also to generate a complete three-dimensional (3D) map of the cell indicating where the proteins are located. To achieve these goals require the involvement of a large number of different fields such as biochemistry, molecular biology and bioinformatics. Many different areas of study are now grouped under the proteomic such as sequence and structural proteomics, expression proteomics, interaction proteomics and functional...

Words: 847 - Pages: 4

Premium Essay

Cloning and Expression of Α-Amylase Gene from Bacillus Subtilis in Pichia Pastoris and Escherichia Coli.

...Cloning and expression of α-Amylase gene from Bacillus subtilis in Pichia pastoris and Escherichia coli. Introduction Enzyme is a type of catalyst that present in living organisms used for many biotechnological functions in various industrial processing. It has a special characteristic that allows the chemical reaction to speed up without being altered, thus significantly improve the industrial productivity (Roy et al. 2012). Among various enzymes available in market, α-amylase has received a special attention in commercial production due to its widely used applications. α-Amylase contributed to 50% of the world enzyme production and has a great importance in many industries such as in food processing, laundry and also in pharmaceutical (Asgher et al. 2007). α-Amylase enzyme acts on α-1,4 glycosidic bonds in starch substrate backbone leading to the formation of soluble maltodextrins, glucose and maltose (Vidyalakshmi et al. 2009). This characteristic is extremely useful especially in industries that require the hydroxylation of starch such as the production of sugar syrups. The α-amylase enzyme can be obtained from various sources such as plants animals and microbes (Ahmed et al. 2011). However, the naturally occurring enzyme is still insufficient to support all the industrial production and therefore, it is crucial to find a new alternative sources, which is cost-efficient and high yield capacity to meet the supply demand (Yin et al. 2003). In industries, the microbial...

Words: 1791 - Pages: 8

Free Essay

Research

...International Journal of Biotechnology. Photon 114 (2015) 444-451 https://sites.google.com/site/photonfoundationorganization/home/international-journal-of-biotechnology Original Research Article. ISJN: 3352-7304: Impact Index: 4.23 International Journal of Biotechnology Ph ton Study of Microbial Beta-Galactosidase Isolated from Fermented Millets Mixture Pooja Jagtap, Humera Bhattiwala and Annika Durve* Department of Biotechnology, Birla College of Arts, Science and Commerce, Kalyan. Dist Thane, 421301, India Article history: Received: 31 March, 2015 Accepted: 07 April, 2015 Available online: 28 September, 2015 Keywords: Beta: galactosidase, ONPG, X: gal, ONPG and X: gal Corresponding Author: Durve A.* Associate Professor Email: annikadurve ( at ) yahoo ( dot ) com Bhattiwala H. Lecturer Jagtap P. Scientific Officer Abstract In the present study microorganisms producing betagalactosidase enzyme were isolated from fermented millets mixture. Three bacterial isolate C2, C4, Y were found to be Gram positive bacilli, Gram negative cocobacili, Gram positive bacilli respectively while isolate SC2 was found to be yeast. These bacterial isolates were identified using various biochemical tests Growth study kinetics of microbial isolates was performed and the optimum pH and temperature was found. Beta-galactosidase enzyme was extracted and the effect of pH, temperature, substrate variation and incubation period on enzyme activity was studied. The activity of beta-galactosidase for all...

Words: 4190 - Pages: 17

Free Essay

Agriculture Trend in Bangladesh

...Agricultural Research Priority : Vision- 2030 and beyond Sub-sector: Livestock Professor Dr. A.M.M. Tareque And Dr. Shah Md. Ziqrul Haq Chowdhury Bangladesh Agricultural Research Council Farmgate, Dhaka April 2010 Research Priority in Agriculture and Vision Document-2030 and beyond Table of Contents Sl. No. 1 2 3 4 Subject Methodology/Work plan Terms of Reference (TOR) of the Group Leader Executive Summary Vision Document 2030 and beyond: Livestock Research in Bangladesh Background Review of the past Plans Targets: Achievable goals of livestock sub-sector under Vision 2021; Bangladesh for Resolution of Crisis and a Prosperous Future” Problems/Constraints Research Areas Commodity wise Research Priority Large Ruminants (Cattle and Buffalo) Small Ruminants (Goat and Sheep) Poultry Common to livestock health and production Hill Research Conclusion References Page No. 2 2 3 5 5 6 7 5 11 13 8 9 10 11 12 13 14 15 16 17 13 15 18 19 24 27 30 31 33 33 1 Research Priority in Agriculture and Vision Document-2030 and beyond Methodology/Work plan Twelve Experts Team have been formed in BARC in connection with the preparation of Vision Document–2030 and beyond vide letter No.ARC/P&E/103/2008/1540, dt. 29-10-09. Livestock Sub-sector group composed of Professor Dr. A.M.M. Tareque, as Group Leader and Dr. Shah Md. Ziqrul Haq Chowdhury, CSO (Livestock), BARC, as Member-Secretary. The work started with the convening of a day long workshop on SPGR priority setting...

Words: 11793 - Pages: 48

Premium Essay

Ptc Genotype Determination Basing on Dna Samples

...Andrei Vasiliev 01040632 PTC Genotype Determination basing on DNA Samples (Obtained From Individuals with known PTC Test result) that are incubated with Restriction Enzyme (Hae III) Abstract: The ability to taste the bitter compound phenylthiocarbamide (PTC) and related chemicals is bimodal, and all human populations tested to date contain some people who can and some people who cannot taste PTC. Why this trait has been maintained in the population is uncertain but this polymorphism may influence food selection, nutritional status or thyroid metabolism. The gene product that gives rise to this phenotype is unknown, and its characterization would provide insight into the mechanism of bitter taste perception. Although this trait is often considered a simple Mendelian trait, i.e. one gene two alleles, a recent linkage study found a major locus on chromosome 5p15 and evidence for an additional locus on chromosome 7. The development of methods to identify these genes will provide a good stepping-stone between single-gene disorders and polygenic trait. [5. Guo SW, Reed DR.Department of Pediatrics, Medical College of Wisconsin, Milwaukee, USA.] Introduction:The genetic taste phenomenon of PTC was discovered in 1931 when a DuPont chemist named Arthur Fox accidentally released a cloud of a fine crystalline PTC. Standing by his side colleague complained about the bitter taste, while Dr. Fox, who was closer and should have uptaken a strong dose, tasted nothing. Fox then continued...

Words: 2373 - Pages: 10

Premium Essay

Protein Assay by Biuret and Folin Methods

...BASIS OF LIFE PROTEIN ASSAY BY BIURET AND FOLIN METHODS Student Number and Name:1316740 Katerina Koupa | Name of Lab Instructor: Dr. J. Walton | Date of the experiment:06/02/2014 | Date of Report Submission:20/02/2014 | (Fig.1) Biuret Reagent Structure ABSTRACT: This report describes the use of the Biuret and Folin methods of protein assay, to verify the protein concentrations of two unknown solutions. To calculate the protein concentrations of the unknowns, nineteen samples were passed through a spectrophotometer which twelve of them had a known protein concentration and there absorbance levels were found. Then a calibration graph was plot that determines the concentration levels of the unknown samples. INTRODUCTION: One of the most typical procedures applied by lab scientist is the protein quantification. Finding protein concentrations in solutions is important in many ways. First of all detecting the levels of a protein in body fluids can result in identifying various diseases. Also protein verification is required for characterization and purification of enzymes. Because of the significance of protein assays laboratories perform this techniques on almost a daily basis. For this experimental procedure, the Biuret and Folin methods were used to determine the protein concentrations of two unknown samples. First the Biuret method (Fig.1) is based on the appearance of peptides bonds in proteins. When cupric ions (Cu2+)...

Words: 2215 - Pages: 9

Premium Essay

Enzyme Lab Report

...industrial processes (e.g. temperature, salts and pH).Enzymes can be produce by using various microorganisms but in laboratory bacteria and fungi are the common source of enzymes. The main draw back with production of bacterial enzymes is the requirement of cost intensive procedures for separation of enzymes from cells, on the other hand enzyme from fungal origin offer an advantage of separation of mycelium by simple filtration. Besides, the fungus can be grown on inexpensive substrates. Aspergillus species is mostly known for its role as a pathogen. However, they also play important roles in the industry and our environment. A few examples are the A. Niger and A. oryzae species. For the production of enzymes for industrial use, isolation and characterization of new promising strain is a continuous process. They are generally produced by using submerged fermentation due to its apparent advantages in downstream in spite of the cost intensiveness for medium components. The purpose of this work was to study the production of hydrolytic enzymes by aspergillus Niger in submerged fermentation and to determine their activities. 2. Material and Methods: 2.1 Microorganism: A. Niger contributes to the biotechnology industry by providing useful enzymes for the food industry such as amylases, lipases, cellulases, xylanases and proteases. This common mould is involved in many industrial processes including enzymes (e.g. amylases), commodity chemicals (e.g. citric acid) and foodstuffs (e.g. soy...

Words: 1870 - Pages: 8

Premium Essay

Pepper Seed Dna Extraction

...Pepper Seed DNA Extraction Biochem lab: CHE 452L marisol gomez Pepper Seed DNA Extraction Biochem lab: CHE 452L marisol gomez 2015 2015 INTRODUCTION The jalapeno is a member of the capsicum family, along with many other peppers. The usual methods for characterization of different pepper species are based on their morphological and physiological traits, however this many not always be enough. For peppers, their traits are influenced by things like their genotype or their specific environment. Genomic markers can allow for a more direct comparison of closely related individuals (Ansari and Khan, 2012). In our case we focus on DNA extraction. The two basic parts of a DNA extraction procedure include the breaking of the cell walls to expose the DNA and the use of enzymes to remove contaminants. The DNA is analyzed for purity by taking the absorbance. The pure DNA is then visualized by gel electrophoresis. The DNA extraction of plant seeds is difficult because of their cell wall. The method used to break the cell wall includes grinding the seeds with liquid nitrogen. The addition of DNAzol is used to isolate genomic DNA (Chomczynski et al. 1997). Restriction enzymes are necessary to fragment patterns of the DNA and in turn making it easier to analyze the DNA through gel electrophoresis. BACKGROUND The purpose of our experiment is to extract the DNA from pepper seeds to be able to compare and contrast the similarities in their DNA. The extraction of DNA from a plant...

Words: 2410 - Pages: 10

Free Essay

Functional Characterization of Chitin and Chitosan

...Current Chemical Biology, 2009, 3, 203-230 203 Functional Characterization of Chitin and Chitosan Inmaculada Aranaz, Marian Mengíbar, Ruth Harris, Inés Paños, Beatriz Miralles, Niuris Acosta, Gemma Galed and Ángeles Heras* Department of Physical Chemistry II, Faculty of Pharmacy, Institute of Biofunctional Studies, Complutense University, Paseo Juan XXIII, nº 1. Madrid 28040, Spain Abstract: Chitin and its deacetylated derivative chitosan are natural polymers composed of randomly distributed -(1-4)linked D-glucosamine (deacetylated unit) and N-acetyl-D-glucosamine (acetylated unit). Chitin is insoluble in aqueous media while chitosan is soluble in acidic conditions due to the free protonable amino groups present in the D-glucosamine units. Due to their natural origin, both chitin and chitosan can not be defined as a unique chemical structure but as a family of polymers which present a high variability in their chemical and physical properties. This variability is related not only to the origin of the samples but also to their method of preparation. Chitin and chitosan are used in fields as different as food, biomedicine and agriculture, among others. The success of chitin and chitosan in each of these specific applications is directly related to deep research into their physicochemical properties. In recent years, several reviews covering different aspects of the applications of chitin and chitosan have been published. However, these reviews have not taken into account...

Words: 10560 - Pages: 43

Free Essay

Phytochemical Studies and in Vitro Cytotoxicity Screening of Piper Betle Leaf (Pbl) Extract

...have been used in Chinese and Indian folk medicine for centuries and recently been proposed to be used as a chemopreventive agent because of its anti-oxidant activity. In the present paper, Piper betle leaves were standardized for stomatal index, vein islet and vein termination numbers, palisade ratio, UV fluorescence and different ash values. The Piper betle leaves are earlier reported to possess anticancer potential. Hence, the aqueous extract of the leaves was subjected to cytotoxicity studies on Hep-2 cell line using MTT and SRB assays. The mean CTC50 was found to be 96.25 µg/mL, which proved the potent cytotoxicity and hence, the probable anticancer property of the selected extract. Key words: Betel Leaf Anticancer Phytochemical characterization and its leaves, with a strong pungent and aromatic flavour, are widely consumed as a mouth freshener [4]. The leaves are credited with wound healing, digestive and pancreatic lipase stimulant activities in the traditional medicine [5]. During our exploration of non-toxic and affordable herbal medicinal formulations, the PBL extract and its constituent phenolics were found to show impressive anticancer activities [6-8]. The deep green heart shaped leaves of betel vine are popularly known as Paan in India [9, 10]. MATERIALS AND METHODS Collection of...

Words: 2882 - Pages: 12

Free Essay

Determination of Insecticidal Activity of Kenyan Bt Isolates Against the Spotted Stem Borer, Chilo Partellus

...Kenya Agricultural Research Institute, NARL. P.O. Box 14733-00800 Nairobi 2 Chemistry Department, Kenyatta University, Correspondence: kasina.j@gmail.com 2 Abstract The delta – endotoxin crystals containing insecticidal Cry proteins were isolated from 20 unidentified local Bacillus thuringiensis Berliner (Bt) isolates and a reference isolate Bt aizawai (Bta) using froth floatation and low speed centrifugation at KARI Biotechnology laboratories, NARL. The total protein was quantified using the Bradford assay method and protein yield from the nutrient broths was estimated at 3.11 mg/ml ±0.938 mg/ml of nutrient broth culture with a purity level of 54.8% ±15.3% of the protein pellet. Leaf – dip bioassay was used to determine the efficacy of the isolates against Chilo Partellus (Swinhoe), a wellestablished and invasive stem borer in Kenya. Among the isolates, Bt 44 and Bt 48 had the most potent endotoxin crystals to 1st instar C. partellus larvae. In addition, the efficacy of these two isolates was not statistically different (P>0.05) from that of Bta based on LT50 values. The findings show that these two locally available unidentified Bt isolates could be used in management of C. partellus and their characterisation (Cry protein) could aid in their utilization. Key words: Bt efficacy; Bt isolates; LT50; LC50; Introduction Maize (Zea mays L.) is the most important staple food crop in Kenya as well as in the subSahara Africa. However, farmers are yet to attain...

Words: 4095 - Pages: 17

Premium Essay

Pearls Before H1N1

...B and C viruses are almost exclusively isolated from man, although influenza C virus has also been isolated from pigs and influenza B has recently been isolated from seals." 4. ^ Bouvier NM, Palese P (September 2008). "THE BIOLOGY OF INFLUENZA VIRUSES". Vaccine 26 Suppl 4 (Suppl 4): D49–53. doi:10.1016/j.vaccine.2008.07.039. PMC 3074182. PMID 19230160. http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pmcentrez&artid=3074182. 5. ^ Kimura H, Abiko C, Peng G et al. (April 1997). "Interspecies transmission of influenza C virus between humans and pigs". Virus Research 48 (1): 71–9. doi:10.1016/S0168-1702(96)01427-X. PMID 9140195. 6. ^ a b Matsuzaki Y, Sugawara K, Mizuta K et al. (February 2002). "Antigenic and Genetic Characterization of Influenza C Viruses Which Caused Two Outbreaks in Yamagata City, Japan, in 1996 and 1998". Journal of Clinical Microbiology 40 (2): 422–9. doi:10.1128/JCM.40.2.422-429.2002. PMC 153379. PMID 11825952. http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pmcentrez&artid=153379. 7. ^ Lynch JP, Walsh EE (April 2007). "Influenza: evolving strategies in treatment and prevention". Semin Respir Crit Care Med 28 (2): 144–58. doi:10.1055/s-2007-976487. PMID 17458769. 8. ^ a b c "Swine Influenza". Swine Diseases (Chest). Iowa State University College of...

Words: 8637 - Pages: 35

Premium Essay

Malunggay

...Moringa oleifera: A Review of the Medical Evidence for Its Nutritional, Therapeutic, and Prophylactic Properties. Part 1. Jed W. Fahey, S c.D. PEER REVIEWED Johns Hopkins School of Medicine, Department of Pharmacology and Molecular Sciences, Lewis B. and Dorothy C ullman C ancer C hemoprotection C enter, 725 N. Wolfe Street, 406 WBSB, Baltimore, Maryland, USA 21205-2185 Email: jfahey@jhmi.edu Trees for Life Journal 2005, 1:5 The electronic version of this article is the complete one and can be found online at: http://www.tfljournal.org/article.php/20051201124931586 Receiv ed: September 15, 2005; Accepted: November 20, 2005; Published: December 1, 2005 Copyright: ©2005 Jed W. Fahey This is an Open Access article distributed under the terms of the C reative C ommons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Moringa appears to be a nutritional and medicinal cornucopia. The author, a Western-trained nutritional biochemist who has studied some of Moringa’s phytochemicals for almost a decade, gives a brief commentary and extensive references, and presents a table introducing some of the tree’s most intriguing features. This is the first article in a series, and will be followed by more detailed analysis of some of the strongest claims made regarding this edible plant. Vie Art Abstract Moringa oleifera, or the horseradish tree, is a pan-tropical species...

Words: 10491 - Pages: 42