The present study was carried out to clone a glycoside hydrolase GH 13 family enzyme from Thermotoga petrophila into Escherichia coli and characterization of the recombinant enzyme. After amplification of the GH 13 family gene of Thermotoga petrophila, it was cloned in E. coli DH5α by using pTZ57R/T as a cloning vector. Screening of positive clones was performed by colony PCR, double digestion of recombinant pTZ57R/T containing GH 13 family gene with NdeI and HindIII as well as by sequencing of cloned gene. Expression of GH 13 family gene was checked in E. coli BL21 (DE3) by using pET 21a (+) as an expression vector. The growth conditions i.e temperature, pH, effect of IPTG and time of induction and optical density of culture at the time of induction were optimized for maximum expression of GH 13 family gene. Various other fermentation parameters like size of inoculum, agitation rate, effect of different media, aeration rate and dissolved oxygen were also studied for maximum expression of cloned gene. Purification of the recombinant GH 13 family enzyme was carried out by heat treatment followed by ion exchange chromatography with 34.6-fold purification having specific activity of 126.31 U mg−1 and a recovery of 56.25 %. Molecular weight of the purified GH 13 family enzyme, 70 kDa, was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme was stable at 100 °C temperature and at pH of 7.0. The enzyme activity was increased in the presence of metal ions especially Ca+2 and decreased in the presence of EDTA indicating that the α-amylase was a metalloenzyme. However, the addition of 1 % Tween 20, Tween 80 and β-mercaptoethanol constrained the enzyme activity to 87, 96 and 89 %, respectively. No considerable effect of the organic solvents (ethanol, methanol, isopropanol, acetone and n-butanol) was observed on enzyme activity. Line-weaver Burk plot showed Km and Vmax values of 12.35 mM and 25.839 U/ml/min, respectively. Thermodynamic parameters for hydrolysis of soluble starch were found to be Ea=28.445 KJ/mol, ΔH= 34.12 KJ/mol, ΔS= -6.7 KJ/mol and Q10=0.47. Conserved domain analysis of GH13 family protein showed that it it comprises of three conserved domian: AmyAc_MTase (maltosyltransferase), domain of unknown function and AmyA (glycosidase). Homology modelling of GH13 family gene was carried out using 2 templates 1gjuA and 4gkl.1. Enzyme-substrate docking of GH 13 family enzyme was carried out by using maltotriose and dextrin as substrates. 76% desizing of cotton cloth with purified recombinant GH13 family enzyme was achieved at optimized conditions (with 150U/ml enzyme in a buffer of pH 7.0 at 80ºC after 60 min of incubation). In the light of all results obtained in this study it is concluded that this recombinant GH13 family enzyme could be used as beneficial candidate for textile industry.
Before the advent of Islam, women were living in worst conditions during the Era of Ignorance. Daughters were buried alive. Women were openly sold in the market. A woman was just a source of enjoyment for males and nothing else. She had no basic rights. Even she had no share in inheritance. In human history, it is Islam that for the first time gave due respect to woman by providing her basic rights and proper status. Islam believes in equality of men and women – ‘Equality’ does not mean ‘to become identical’. Women’s rights in Islam pertain to Spiritual, Economic, Educational and Legal Rights. A woman can adopt any profession, she wants. Thus, she has got full freedom to live a happy and peaceful life. Unfortunately, there are many people who believe that because Islam gives men the right of having authority over their wives, therefore, men are superior to women and that men have priority over women in all matters. The fact is that according to Islamic teachings, men are not superior to women; and so women are not superior to men. There is no gender discrimination in Islam. Allah Almighty states very clearly in the Holy Quran that superior in His eyes are only those who have more taqwa (fear of Almighty Allah).
The dissertation discusses the censorship of visual pleasure and gaze in early and post-1979 Pakistani films in view of the issuance dates of the Motion Picture Ordinance (1979) and the Censorship of Film Rules (1980). It analyzes films and censor certificates of films considering that the Central Board of Film Censors (CBFC) has been censoring films for one nation in both the eras. However, contemporary films have replaced sexually passive girls of early films with bold and beautiful contemporary women, reflecting a change in the trends of censorship. The dissertation evaluates visual pleasure and gaze at female representations in films from both the eras. The dissertation reviews film scholarship to understand the relationship of cinema to state: stability of state and the threat of public uproars over issues of decency, morality, and sexuality in films. It also reviews the coverage of Pakistani films and industry in Dawn daily and other literature over the past decade to evaluate the performance of Pakistani cinema under different political regimes, its decline, present status and issues. However, its main focus remains on the censorship of visual pleasure in Pakistani films. The study uses critical theories to discuss male fascination and anxiety with the female form, social formations, and epistemology to examine the shift in the reality and representation of women to opposite poles in the pre-and-post-Zia films. The study uses triangulation of methods to analyze the films and the excisions as noted on the censor certificates of films by the CBFC in the pre-and-post-1979 eras. It evaluates the consistency in the vision and approach of the machinery of the state in implementing the Pakistani Cinema 23 censorship policy to sustain cultural norms, religious harmony, and peace in the country over a period of time. It evaluates various aspects of historical changes in interpreting and understanding Censorship of Film Rules about the political system, national identity, decency, morality, and sexuality, as both the legislation and film are reflection of wider society as subjective exercises. The review of Pakistani literature comprising the statistical data of films does not prove the authenticity of a general perception that General Muhammad Zia-ul-Haq’s martial law regime and forced Islamization caused the decline of Pakistani cinema. The study of the censor certificates reflects that the CBFC excised more visual pleasure (dances-vulgar body movements and sexual innuendoes) from films produced in the pre Zia era and less from films produced in the post-Zia era. A change in ethics and mores of society is visible in images that were puritanical in early days and morally and sexually liberal later. The study explores élite gazing at traditional girls of the early films as opposed to the common gazing at free women who are open with their emotions and sexuality in contemporary films. The study finds that the early Pakistani films promoted the eastern cultural norms of Urdu speaking élites of Delhi and Lucknow against sexual boldness of westernized antagonists. The representations have changed tremendously over the decades replacing the shy women with the sexually bold and beautiful Punjabi heroines who act like the antagonists of the past. The study reflects changes in the censorship of visual pleasure in films though CBFC still censors the films as per rules.