Pakistan’s Stance on the ‘War on Terror: Challenging the Western Narrative

The tragic events of 11 September 2001 allowed the United States to reframe its pursuit of global hegemony as ‘War on Terror’ which is styled on Islamophobic rhetoric and action. To counter this campaign of Islamophobia, Pakistan has adopted a consistent and well-planned stance. The essential contours of Pakistan’s stance on the ‘War on Terror’ are the need to: condemn terrorism in all its forms and manifestations; defend Islam and Muslims; support the right of self-determination of oppressed people particularly, Palestinians and Kashmiris; respect international law; address the root causes of terrorism; and promote peace and harmony among cultures, civilizations and followers of diverse religions all over the world through promotion of a robust dialogue and  criminalization of  defamation of religions.

Production of Bacterial Pectinase Using Submerged Culture for Various Applications

Pectinases are pectin degrading enzymes and are naturally produced by plants, animals and microorganisms. Their major source of production at industrial scale is from microorganisms especially Bacillus sp, Aspergillus sp. and yeast sp. which are generally regarded as safe. Aspergillus sp. generally produce acidic pectinases which are used in the food and beverage industry for the extraction and clarification of fruit juices and maceration of vegetables for production of purees and pastes. Bacillus sp. are usually capable of producing alkaline pectinases which have diverse functions and are in use in many industrial processes, successfully substituting the use of harsh chemicals which not only causes the deterioration of product quality but also the deterioration of environment. Pectinases from Bacillus sp. are generally active at broad ranges of pH and temperature and due to this reason, they are multi-functional enzymes. Pectinases account for more than 10% of the industrial enzymes market and they constitute 25% of the global food enzymes market. The present study was concerned with the search for a novel bacterial isolate for the lab scale production of pectinase (Polygalacturonase). Keeping in view the increasing demand of pectinase, specially its need in Faisalabad, a textile city of Pakistan, isolation of new hyper producer bacterial strains locally is an easy and cheap way of getting the desirable products at low cost. Therefore, isolation of new strains for industrial enzyme production has been, and will be, a part of research. This method alone can also provide raw material for further research such as enzyme engineering or molecular directed evolution. Pectinase positive cultures were isolated using PSAM, the medium that is able to grow and differentiate pectin consuming bacteria from others. The pectinase producing bacteria form clear halos around their colonies while others do not form any clear zones. For the identification of hyper producer strains, colony PCR was done for 16S rRNA analysis. The reason to use the 16S rRNA gene for identification purposes is that there is a large database of DNA sequences available for the gene from the widest range of microbial species as compared with any other genetic target. The selected bacterial isolate NS1 (source of pectinase enzyme) was identified based on PCR amplification of 16S rRNA and for this purpose the amplified product was electrophoresed in agarose gel against a known species of Bacillus licheniformis. The 16S rRNA sequencing confirmed the Bacillus status of the strain NS1 and the nucleotide sequence BLAST results showed 98% similarity of strain NS1 having Accession No. KX765286 with few species of Bacillus licheniformis. The growth conditions of the newly isolated Bacillus licheniformis strain were investigated using submerged fermentation to understand the fermentation behaviour of the microorganism and the pattern of pectinase production by it. The growth of the organism and enzyme production by it was investigated using some local agrowastes such as wheat bran, gram bran, citrus peel, apple pomace, carrot pomace and peanut shells as carbon sources. Among these agrowastes citrus peel powder at 2.5% concentration proved as best substrate for pectinase production followed by wheat bran. Among various nitrogen sources investigated for their role in pectinase production, organic sources such as tryptone and yeast extract gave better results than inorganic nitrogen sources. Among inorganic sources Diammonium hydrogen phosphate gave more pectinase units than other inorganic nitrogen sources. Physical parameters like pH, temperature, inocula size and incubation period for high yield of pectinase in submerged fermentation were optimized by using Response Surface Methodology. Which is an efficient tool for increasing product yield many folds in short time due to limited number of experiments and lab work. In the present study, the yield of pectinase was increased 5.6 fold that optimization produced 219U/mL as compared to one variable at a time method which produced only 38.86U/mL. Several purification methods were evaluated to observe that which one is more advantageous and cost effective in the present study for pectinase purification. Three methods of protein purification (aqueous two phase purification system, macro-affinity ligand facilitated three phase partitioning and gel filtration chromatography) were used in the present study and macro-affinity ligand facilitated three phase partitioning were found to give high purification of pectinase with purification fold of 13.05. The pectinase from newly isolated Bacillus licheniformis showed some novelty in characteristics as compared to most of the pectinases produced by other species. Although its optimum activity was achieved at a temperature of 70ºC in glycine buffer pH 8 but it also showed considerable activity (26.75U/mL) even at 100ºC in phosphate buffer pH 7. Addition of 15mM CaCO3 to the enzyme assay mixture increased the pectinase activity by 3.1 fold and addition of chloroform to enzyme assay mixture increased the pectinase activity by 7.45 fold. Surfactants (CTAB, SDS and Triton X-100) increased the pectinase activity many fold as compared to control. Among various sugars investigated for their effect on pectinase activity, sorbitol was found as a stimulator of pectinase activity by increasing its activity by 1.8 fold while glucose, lactose and sucrose inhibited its activity. The pectinase produced in this study was investigated for applications such as oil extraction from sunflower seeds, apple juice extraction and clarification and starch extraction from potatoes. In all of the above applications, the locally produced pectinase enhanced the yield of apple juice, oil and starch several fold as compared to control without the application of pectinase enzymes. Apple juice yield was increased by 2.06 fold due to pectinase treatment while the juice clarification was increased by 1.62 fold. Pectinase application also increased the oil yield some 3.15 fold as compared to oil extraction by water without the addition of pectinase. Its effect on potato starch extraction was also appreciable and 3.95 fold increase in starch yield was observed due to pectinase treatment of potato slurry. All these investigations showed that the low cost pectinase produced by locally isolated microorganisms using low cost agrowastes as nutrient supplements are able to compete with costly commercial enzymes and can bring a revolution in product quality and yield if used by local industries.