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All Journal Brawijaya Law Journal : Journal of Legal Studies Jurnal Teknologi Laboratorium
Dinesh Kumar
Department of Pharmaceutical Sciences, M.D. University, Rohtak India , 124 001.
Chemistry Law, Crime, Criminology & Criminal Justice Other

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Purification and characterization of recombinant Streptokinase expressed in E.coli from Streptococcus equisimilis with N-terminal methionine Vadla, Pavan Kumar; Tummuru, Murali; Kumar, Dinesh
Jurnal Teknologi Laboratorium Vol 8 No 1 (2019): 2019 (1)
Publisher : POLTEKKES KEMENKES YOGYAKARTA

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (397.321 KB) | DOI: 10.29238/teknolabjournal.v8i1.153

Abstract

Streptokinase is a extracellular enzyme which is extracted from strains of beta Hemolytic streptococcus. The enzyme is a non-protease plasminogen activator that activates plasminogen to plasmin and degrades fibrin clot through its specific lysine binding site which is used in thrombolytic therapy. Purification of streptokinase produced from S.equisimilis in E.coli with N-terminal methionine was carried out in 3 Chromatography purification steps, 1) CM-Sepharose-FF at pH 4.2 followed by concentration and dialysis over night with Tris-HCl pH 8.0. Partially purified dialyzed enzyme sample was loaded on to 2) DEAE-Sepharose-FF column. The Purified fractions of DEAE column were pooled and applied on to Sephadex G-100 column. Enzyme purity was confirmed by SDS-PAGE and RP-HPLC.Its biological activity is determined by specific streptokinase assay and characterised the enzyme by Peptide mapping, MALDI-TOF, Isoelectric-focusing and RP-HPLC. The isoelectric point (pI) of streptokinase is around 4.98.The results of characterization shows that it contains two forms (Isomers) of streptokinase expressed in E. coli which was analyzed by RP-HPLC and chromogenic assay. The variation is formed by isomer-1 in which 85% of Streptokinase expressed without methionine (85000IU/mg) and Isomer-2 in which 15% of streptokinase expressed with methionine (nil activity) in E. coli. This phenomenon shows that the presence and absence of methionine in isomers of streptokinase varying the catalytic activity of the enzyme.
Need for Revamping Information Technology Laws in India Walia, Ivneet Kaur; Kumar, Dinesh
Brawijaya Law Journal : Journal of Legal Studies Vol 8, No 2 (2021): State Administration Role in Establishing Constitutional Obligation
Publisher : Faculty of Law, Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21776/ub.blj.2021.008.02.03

Abstract

Homo Sapiens have a distinct characteristic of being superior to other creatures. They owe this superiority not only because they have the power to reason and rationalize but also because they have a tendency to organize themselves as a congregation, which can work in a group at a large scale. The human instinct to improvise on its own inventions, have today resulted in mutation of a world from the stage of abacus to the era of Robotics. For the sake of avoiding chaos and maintaining the sovereignty, every nation strives to eradicate the fear of dominance by the selected elite and hence the need of regulations and the law. The aim of the paper is to highlight the grey areas and limitations existent in Information Technology Laws and focusing on the emerging domains of cyberspace. Its also aims to draw the attention by the policymakers and the legislators to understand the need for amending the Information Technology Act, 2000 for including legal provisions related to emerging issues in cyber space. The analytical research method is used to collect the data based on a systematic review of the existing sources of information and involved qualitative research to analyze the information. The conclusion and suggestions of this paper will definitely be helpful in either drafting or amending a comprehensive law relating to IT keeping in view the evolving technologies and their applications.
Co-Authors Tummuru, Murali Vadla, Pavan Kumar Walia, Ivneet Kaur