iman p. maksum et al /j. pharm. sci. & res. vol. 9(6), 2017, 1007...

Iman P. Maksum et al /J. Pharm. Sci. & Res. Vol. 9(6), 2017, 1007-1016

1007


1008

EDITORIAL BOARD

Chief Editor

Mrs. R. Radhika,Pharmainfo Publications, India

Editorial Board Members

Dr. Chitra Telang,Senior Scientist, Pharmaceutical R&D,a Boehringer Ingelheim Pharmaceuticals, Inc.1 Eastview Lane,Brookfield, CT 06804, USA

Dr. Mustafa OSKAY,Biology Dept., Faculty of Sciences and Arts,Celal Bayar University, Campus of Muradiye,45010, MANISA/ Turkiye

Dr. Marco Cosentino,Associate Professor of PharmacologyFaculty of MedicineDepartment of Clinical MedicineSection of Experimental and Clinical PharmacologyUniversity of InsubriaVia Ottorino Rossi n. 921100 Varese VA – Italy

Dr Bhupinder Singh Bhoop,Professor (Pharmaceutics & Pharmacokinetics)Univ Inst Pharm Sci (UGC Center of Adv Studies)Dean (Alumni Relations), Panjab UniversitySector 14, Chandigarh 160014, India

Dr. Raghavendra Kulkarni,ProfessorBLDEA's College of PharmacyBLDE University, BIJAPUR 586103Karnataka, India.

Dr. Zhaofeng Ding,Senior Scientist / Parenteral Technologies


1009

BD Technologies21 Davis Drive, Research Triangle Park, NC 27709 USA

Dr. Vikas Anand saharan,Assistant Professor in PharmaceuticsIncharge, Committee for Higher Education Guidance (CHEG),Institute of Pharmaceutical Sciences & Drug Research,Seth GL Bihani SD College of Technical Education,Sri Ganganagar 335001, Rajasthan, India

Dr. Angelo SpadaroDepartment of Pharmaceutical SciencesUniversity of CataniaCittà UniversitariaViale Andrea Doria 695125 Catania, Italy

Prof. Dr. Maria Laura ColomboMedicinal Plant BiologySchool of PharmacyUniversity of TurinItaly

Dr.Khawaja Tahir Mahmood,Pharmacist ,DTL,Punjab, Lahore /Visiting Associate Professor,Department Of Pharmacy, Lahore College For Women University, Lahore,Pakistan

Dr. Sree Harsha N.College of Clinical PharmacyKing Faisal University, Al-Ahsa, Saudi Arabia

Dr. M. Amarnath Satheesh,Lab C6, 11937 U.S HWY 271University of texas health center at tylerTyler, Tx 75 USA

Dr. R.Bhaskararao Rayini,Scientist, Capricorn Phamra IncFrederick, Maryland ,USA


1010

ISSN 0975–1459

Volume 9, Issue 6 June 2017.

1. Preparation of Non- toxic dose of aqueous extract of Strychnine from the stem pieces of Strychnos-nux-vomica.Pages:762-765V.Varaprasad, Dr R. Bharath Kumar

Abstract | PDF (760 K)

2. Method of Increasing Ecological and Consumer Qualities of Meat and Intensification of the DigestiveMetabolism Processes in Broilers Grown in Technogeneous AreasPages:766-770Lada A. Vityuk, Sergei I. Kononenko, Aleksandr V. Yarmoc,Irina R. Tletseruk, Lidia V. Chopikashvili


3. Investigation of Early Maladaptive Schemas in Patients with Bipolar Disorder Compared to HealthyIndividualsPages: 771-774Shahla Khosravi, MohammadIsmaeil Ebrahimi, Arezoo Shayan, Mohamad Reza Havasian, FarkhondehJamshidi


4. Criteria for Revealing the Early Preclinical Signs of Benzene IntoxicationPages: 775-779Ragim Almamed Orujov, Rana Anver Jafarova


5. Impact of the Breed-Specific Characteristics on the Metabolism and Heavy Metal Accumulation intheOrgans and Tissues of CalvesPages: 780-784Rustem B. Temiraev, Sergei I. Kononenko, Soslan G. Kozyrev, Angelika A. Baeva, Larisa A.Bobyleva,Arthur L. Kalabekov


6. Gastro protective Effect of Pentagalloyl Glucose on Aspirin Induced Gastric Mucosal Ulcer inComparison with Omeprazole, Famotidine and MelatoninPages: 547-551Fatma A. Moharram, Rabab A. El Dib, Lobna F. Wahman, Siham M. El-Shenawy, Sally El-Awdan


7. An Overview of the most Important Medicinal Plants with Anti-Toothache Property based onEthnobotanical Sources in IranPages: 796-799Somayeh Ghamari, Reza Mohammadrezaei-Khorramabadi, Mahnaz Mardani, Somayeh Shahsavari



1011

8. Identifying medicinal plants affecting the teeth from the Southern District of Ilam province, IranPages: 800-803Somayeh Ghamari, Saber Abbaszadeh, Mahnaz Mardani, Somayeh Shahsavari


9. An Overview of the most important Medicinal Plants used as Mouth FreshenerPages: 804-807Somayeh Ghamari, Reza Mohammadrezaei-Khorramabadi, Mahnaz Mardani, Somayeh Shahsavari


10. Organizational, Clinical and Social Aspects of Age-Related Sarcopenia Diagnostics in Patients withObesityPages: 808-811Aleksandr Aleksandrovich Zarudskiy, Elena Aleksandrovna Perutskaya, Dmitry Nikolaevich Perutskiy,Natalya Yurevna Stasevich, Elgudzha Yakovlevich Nemsstveridze


11. Synthesis and Antimicrobial Activity of some Methyl -4-(Benzo[D] Thiazol-2yl)Phenylcarbamodithioate Amine DerivativesPages:812-816Laliteshwar P. Singh, Dr. O.P.Tiwari, Brijyog


12. Biological Substantiation of Application of the Coniferous-Energy Supplement in Feeding of HeifersPages: 817-821Yuri Nikolayevich Prytkov, Anna Aleksandrovna Kistina, Vasiliy Pavlovich Korotkiy, Viktor AnatolyevichRyzhov, Viktor Ivanovich Roshchin


13. Sodium Valproate Induced Hyperammonemia with Normal Liver Function: A Case ReportPages:822-823Teena C Abraham, Greeshma James, Anjana Thomas, Lakshmi R


14. Multivariate Calibration Technique for the Spectrophotometric quantification of Zaleplon in bulkdrug and pharmaceutical formulationsPages: 824-829Kokilambigai K. S., Seetharaman R., Kavitha J., Sowndaravel P., Lakshmi K. S.


15. Entomopathogenic Fungi: An Effective Biocontrol Agent for Management of Insect PopulationsNaturallyPages: 830-839Digvijay Singha, Tanveer Kour Rainab, Joginder Singha


16. Oral and Maxillofacial approach for the treatment of Obstructive Sleep Apnea - ReviewPages:840-844Gadicherla Srikanth, Evit Rajan John, Kalyana Chakravarthy Pentapati



1012

17. The Treatment for Common Cold in Out-Patient Setting in Health Care Service Units Provided by aCoal Mining Company in IndonesiaPages: 845-849Erna Kristin, AlfiYasmina


18. Optimization of fermentation and nutritional parameters for the production of L-Methionine byC.glutamicum using agriculture productsPages: 850-856A. Venkata Narayana and A.V.N.Swamy


19. Pharmacology and Bio-medicinal properties of Chitin and its derivativePages: 857-859Nia Lopez,Athira P.N, Pooja Ajeesh, Anjali.S.Nair, Ankitha.L, Anu priya


20. Stability Indicating RP-HPLC Method Development and Validation of Tolterodine Tartrate and itsDegradant in its Capsule Dosage FormPages:860-866Urvi S, Nilesh P, Ashok P, Amit V, Ajay P


21. Liposomal Amphotericin B Induced Pancytopenia- A Rare Case ReportPages: 867-868Jeslyn Mary Philip, Meenu Vijayan


22. Prevalence of Dental Developmental Anomalies among Men and Women and its Psychological Effectin a Given Population.Pages: 869-873Harsha.L, Dr. M P Brundha


23. Influence of the Chlorophytum Comosum Leaves Hydroalcoholic Extract on Some Representativesof Intestinal Microflora of RatsPages: 874-877BONDAREVA N.I., TIMCHENKO L.D., DOBRYNYA Y.M., AVANESYAN S.S., PISKOV S.I.,RZHEPAKOVSKY I.V., KOZLOVA M.A, ARESHIDZE D.A, LYHVAR A.V.


24. Selection of Significant Lifestyle Risk Factor of Cancer by Hybrid X- Bar – DEMATEL-TOPSISMethodPages: 878-885Priyanka Majumder, Apu Kumar Saha, Mrinmoy Majumder



1013

25. Rheumatoid Arthritis and Chronic Periodontitis – A Disease Link.Pages: 886-890DR.T.SIVASANKARI, DR.T. RAMAKRISHNAN, DR.B.ARUNA


26. Development of Strength Capacity of Students on the Basis of Static-Dynamic ExercisesPages:891-896Larisa Vladimirovna Byankina, Vladimir Vasilyevich Byankin, Irina Mikhailovna Vorotilkina, MariaMikhaylovna Prokopyeva, Aya Petrovna Bugaeva


27. Hematological and Epidemiological Study for Patients Infected With ScabiesPages: 897-900Saleem Khteer Al-Hadraawy, Hanin Bahaa Hessen


28. Study of some Biochemical parameter in patients infected with Giardia lambliaPages: 901-903Ali Hassan Abood


29. The Influence of Adequate Iodine Intake on the Level of Stress Resistance and the Capacity of Short-Term Memory in SchoolchildrenPages: 904-909Zulpat Abdulgalimovna Magomeddibirova, Goljan Sahihulislamovna Tazutdinova, Umahani ShamhalovaMagomedkhanova, Alina Balaefendievna Kurbanova, Fatima Ramazanovna Asadulaeva, AzraGitinomagomedovna Abdurakhmanova


30. Contraindications in the Use of Biological Therapies -Active or Recent History of Malignancy andNeurological DisordersPages: 910-913Divyadharsini.V, Roshene.R, Jayanth kumar


31. Prosthodontic Needs in Patient after Tooth Extraction in South Indian PopulationPages: 914-918Anas Bin Rosli, Ashish.R.Jain


32. CoMat: An Integrated Tool for Comet Assay Image AnalysisPages: 919-925Udayakumar Mani A , Pavithrakumari Manickam


33. Interaction Studies of Olive Oil Component Oleocanthal with Anticancer TargetsPages: 926-935HARIHARAN L., OMKAR V. POKHARKAR, VIVEK K. MUKHERJEE



1014

34. The Sterility of Reusable Instruments at Central Surgery Installation’s Room Storage of One of TheHospital in BandungPages: 936-940Insan Sunan Kurniawansyah, Sriwidodo, Soraya Ratnawulan Mita


35. Glucosamine as a Novel Adjunctive Therapy in Symptomatic Oral Lichen PlanusPages: 941-946R.R. Hesen, H.A. Abuel-Ela, I. M. Helmy, M. H. El Sayed, O.M. Ezzatt


36. Development of Ready to Eat Extruded Snacks from Blend of Under-Utilized Legumes and MilletsPages: 947-954Sharmila.B and Athmaselvi.K.A


37. Comparative Study of Anti-Inflammatory Activity of Petroleum Ether Extract of Myxopyrummilacifolium Blume and Ethanolic Leaf Extract of Pamburus missionis Swingle.Pages: 955-957Raveesha Peeriga, K.B.Chandrasekhar


38. Direct Determination of Copper, Lead and Cadmium in the Whole Bovine Blood Using Thick FilmModified Graphite ElectrodesPages: 958-964Tatyana V. Skiba, Alphya R. Tsygankova, Natalya S. Borisova, Kirill N. Narozhnykh, Tatyana V.Konovalova, Olga I. Sebezhko, Olga S. Korotkevich, Valeriy L. Petukhov, and Ludmila V. Osadchuk


39. Platelet Rich Plasma in Periodontal TherapyPages: 965-971Dr.S.Sathya Priya Eshwar, Dr.Dhayanand John Victor, Dr.S.Sangeetha, Dr.PSG.Prakash


40. Formulation and Characterization of Flexible Phosphatydilcholine Vesicles for Systemic Delivery ofPiroxicamPages: 972-985Setareh Barani, Prof. Mushir Ali, Dr. Saima Amin


41. Menstruation Phytotherapy According To Iran Ethnobotanical SourcesPages: 986-990Faeghe Tajallaie-Asl, Mahnaz Mardani, Somayeh Shahsavari, Saber Abbaszadeh


42. Influence of Different Roasting Conditions in the Phenolic Compounds and Antioxidant Capacity ofthe Pistachio NutsPages: 991-993Sara Goudarzi, Tayebehe Asadi Sharifi, Asghar Ahmadi, Sadegh Rezapour, Parisa Sadighara



1015

43. Polycyclic Aromatic Hydrocarbons in Shell and Kernel of Roasted Sunflower SeedsPages: 994-996Behrouz Akbari-adergani, Toran Khaki, Gholamreza Jahed, Reza Ahmadkhaniha,Sadegh Rezapour, Parisa Sadighara


44. Preventive and Detoxicative Action of Probiotics on Metabolism and Consumer Quality of BroilersMeatPages: 997-1001Victoria Viktorovna Tedtova, Mohamed Kadirovich Kozhokov, Larisa Hasanovna Shugusheva, ValentinaNikolaevna Kanukova, Angelica Ahsarbekovna Baeva, Lada Alexandrovna Vityuk


45. Investigation into Chemical Composition of Powdered Lupine SeedsPages: 1002-1006Natalia Aleksandrovna Tarasenko, Elena Aleksandrovna Butina, Evgeny Olegovich Gerasimenko, andYulia Nikolaevna Nikonovich


46. Extracellular Secretion of Recombinant Human Epidermal Growth Factor by Using TrimethylamineN-Oxide Reductase A (TORA) Signal Peptide in Escherichia Coli BL21 (DE3)Pages: 1007-1016Iman P. Maksum, Eriya Utama, Sriwidodo, Toto Subroto


47. Ultrasonic Studies Of Alanine in Aqueous Paracetamol Solutions at Different TemperaturesPages: 1017-1025Rajagopal. K, Roy Richi Renold. G, Mohamed Roshan. M



1016

Extracellular Secretion of Recombinant Human Epidermal Growth Factor by Using Trimethylamine N-Oxide

Reductase A (TORA) Signal Peptide in Escherichia Coli BL21 (DE3)

Iman P. Maksum1* Eriya Utama1, Sriwidodo2, Toto Subroto1

1)Biochemistry Laboratory, Chemistry Department,Faculty of Mathematics and Natural Science, Universitas Padjadjaran, Indonesia, +62227794391

2)Pharmaceutic and Pharmaceutical Technology Department,Faculty of Pharmacy, Universitas Padjadjaran, Indonesia, +62227796200

Abstract Human Epidermal Growth Factor (hEGF) is a protein that helps in the healing process of wounds, in the process of proliferation, migration and cell differentiation. hEGF composed by 53 amino acid residues, with a molecular weight of 6.2 kDa and has a single chain conformation, and resistant to high temperatures up to 70˚C. hEGF is already very widely produced in an industrial scale using Escherichia coli as a host cell, but the majority of hEGF recombinant protein is produced intracellularly, so it has a lot of disadvantages such as; the accumulation of non-functional protein, the amount of protein inclusion bodies are formed as a result of misfolding, and the high rate of degradation due to intracellular protease enzymes. Therefore the purpose of this study was to express recombinant proteins in the extracellular hEGF using a signal peptide TorA, via the Twin Arginine translocation (TAT) pathway, measure the levels of hEGF recombinant protein secreted into the culture medium with the use of a signal peptide TorA via TAT. This research using Escherichia coli BL21 as the host cell to be transformed by using recombinant plasmid pD881-TorA the consensus already containing the signal peptide TorA and also hEGF gene, then expressed by L-rhamnose induction, then the recombinant protein hEGF purified by heat treatment and then the results were characterized using SDS-PAGE, and ELISA. The result is hEGF protein can be secreted into culture medium and periplasm with concentration 0.9625 µg/ mL.

Keywords: Recombinant Protein hEGF, TorA signal peptide, extracellular secretion

INTRODUCTION Human Epidermal Growth Factor (hEGF) is a protein that helps in wound healing, in terms of the process of proliferation, cell migration and cell differentiation. HEGF protein composed of 53 amino acid residues, with a molecular weight of 6.2 kDa and has a single-chain conformation1, hEGF produced by the human duodenum and by the salivary glands2, hEGF is often used in medical and cosmetic fields, for hEGF has the ability to stimulate the growth of various types of cells and accelerate the phase of cell differentiation in tissues. hEGF have broad biological reactions, and quickly spur growth which a variety of skin layers. In general, skin care products just add liquid skin to help moisturize the skin and slow the aging, but it does not improve. Instead hEGF can penetrate the dermis to stimulate the growth of new cells to replace old cells that have been damaged, and serves to control the growth of skin cells. hEGF can help form a protein that can stimulate other tissues to regenerate the cells, so that hEGF is very effective for use as a wound healer protein3. In the industrial world, the production of recombinant proteins is commonly used E.coli as a host cell. Gram-negative bacteria have a very fast growth process, so as to minimize production costs. E.coli is also relatively easy for the recombinant plasmid is inserted into it, so that the gene hEGF can be easily inserted into the plasmid vectors of E.coli. As well as other advantages are, E.coli can reachhigh cell density rapidly4.

But besides these advantages, the use of E.coli as a host cell has several disadvantages, including: (1) the accumulation of protein secretion is non-functional, (2) high chances for the formation of inclusion bodies as a result of incorrect folding (misfolding), and ( 3) the high rate of protein degradation due to their intracellular protease enzymes5. To overcome these drawbacks, the most effective solution is to produce hEGF in extracellular proteins6 by means of a membrane secreting into periplasm or to the culture medium. By using a secretion system in extracellular, can simplify the purification process, avoiding attacks protease enzyme to denature hEGF, and can minimize mis-folding for protein hEGF directed to folding of the right to the presence of protein chaperones before secreted extracellular7. To be able to secrete recombinant protein into the membrane hEGF periplasm or to the culture medium to be used the signal peptide which is fused with the target protein to be passing lane or lanes sec TAT (Twin Arginine Translocation)8. Protein secretion pathways that can make production much more effective, as well as to further optimize the secretion of recombinant proteins used hEGF TorA signal peptide with through TAT. Because the signal peptide TorA and TAT way can secrete proteins are great as Green Fluorescent Protein (GFP) very well, so if the signal peptide TorA and TAT way is used to secrete hEGF small size, it is expected to be much better, because hEGF


1017

iman p. maksum et al /j. pharm. sci. & res. vol. 9(6), 2017, 1007...

Documents