Muhammad Javaid Asad

Bioethanol productions from rice polish by optimization of dilute acid pretreatment and enzymatic hydrolysis

Lignocellulose materials are abundant renewable resource for the production of biofuel from fermentative organism ( Sacchromyces cervesiae ). Rice polish is cheapest and abundant lignocelluloses resource and has potential to produce bioethanol. The main steps for the conversion of biomass into glucose required dilute acid pretreatment, but it also released inhibitory compounds which reduced the ethanol yield. So, attempt has been made to minimize the effect of inhibitory compounds as well as optimized the condition like glucose recovery, xylose solubilization and lignin degradation during dilute acid pretreatment. Maximum lignin degradation, less glucose loss, better xylose solubilization obtained with dilute sulphuric acid 1.5% at 100°C for 30 min. During enzymatic hydrolysis (Novozyme) with 0.75 mL/2 g, enzymatic loads for 72 h gave 4.27mg/mL glucose while with indigenous enzyme load 1 mL/2 g gave 0.953 mg/mL of glucose. Key words: Bioethanol, Dilute acid pretreatment, enzymatic hydrolysis, rice polish.

Phanerochaete chrysosporium IBL‐03 secretes high titers of manganese peroxidase during decolorization of Drimarine Blue K2RL textile dye

A novel indigenous strain, Phanerochaete chrysosporium IBL‐03, with high manganese peroxidase (MnP) activities was used for decolorization of a reactive textile dye, Drimarine Blue K2R, which is used extensively in textile units of Pakistan. The initial experiment was run for seven days with 0.01% (w/v) dye solution prepared in Kirks basal nutrient medium. Samples were removed after every 24 h and the extent of dye decolorization was determined at λmax of the dye. The study revealed that P. chrysosporium caused 65% decolorization of Drimarine Blue K2RL in seven days. By process optimization, 97% colour removal could be achieved in three days using 0.005% (w/v) Drimarine Blue K2RL solution at pH 4.0 and 30 °C in defined Kirks medium with 0.9% (w/v) molasses and 0.2% (w/v) ammonium dihydrogen phosphate added as carbon and nitrogen sources, respectively. Manganese peroxidase was found to be the major enzyme (560 IU/mL) involved in dye decolorization of Drimarine Blue K2RL by P. chrysosporium. The dye adsorption studies showed that the dye initially adsorbed on fungal mats disappeared later on, possibly by the action of MnP secreted by the fungus in secondary metabolism.

Epidemiology of Transfusion Transmitted Infection among Patients with β-Thalassaemia Major in Pakistan

Introduction. Transfusion Transmitted Infections (TTIs) continue to be a major risk in transfusions in many parts of the world. The transfusion-dependent β-thalassaemia patients are particularly at risk of acquiring TTIs. The current study was undertaken to estimate the prevalence of TTIs in transfusion-dependent β-thalassaemia patients. Material and Methods. A cross-sectional study of 1253 multitransfused thalassaemia major patients was conducted in five different centres of Islamabad, Rawalpindi, and Karachi. The study subjects were screened for HIV, HCV, and HBV. The screening was performed at two centres: Department of Pathology, Shaheed Zulfiqar Ali Bhutto (SZAB) Medical University, and Blood Transfusion Services, Jinnah Postgraduate Medical Centre, from July to December 2015. The confirmatory screening was performed by Chemiluminescent Immunoassay (CLIA). Results. Out of the 1253 multiple transfused patients, 317 (25.3%) were infected with TTIs. HCV was positive in 273 cases (21.7%), HBV in 38 cases (3.0%), and HIV in 6 cases (0.5%). Conclusion. HCV was the leading TTI in multitransfused thalassaemia major patients in the study. Presence of HIV in thalassaemia patients is a recent disturbing development in Pakistan. Improved regulation of blood banks including use of internationally or nationally evaluated kits will bring down the incidence of TTIs in transfusion-dependent β-thalassaemia patients. More stringent behavioral and serological pretransfusion screening of blood for TTIs must be implemented in blood banks.

Presence of HCV RNA in peripheral blood mononuclear cells may predict patients response to interferon and ribavirin therapy.

BACKGROUND AND OBJECTIVES Hepatitis C virus (HCV) is considered a hepatotropic virus, but it can replicate in peripheral blood mononuclear cells (PBMCs), which influence the sustained virological response (SVR) of the patients, as well as relapse in successfully treated patients. The main objective of this study was to establish the importance of PBMC HCV RNA detection as a primary test to declare the patient as a responder, and the secondary objective was to investigate the risk of non-SVR or relapse in individuals who showed an end-of-treatment (ETR). DESIGN AND SETTINGS Blood samples were collected after the completion of 6 months of therapy, and they were collected 6 months after the completion of treatment. PATIENTS AND METHODS A total 103 patients infected with the 3a genotype of HCV and those who were treated with interferon-α-2b and ribavirin for 24 weeks were selected. HCV RNA in plasma of at the end of treatment and 6 months after the completion of treatment was determined with the help of quantitative real-time polymerase chain reaction (qRT-PCR). RESULTS Of the 103 patients, 74.8% (number [n]=77) were end-of-treatment responders, while 25.2% (n=26) were nonresponders. Seventy-seven responders were tested for HCV RNA in their PBMCs. The HCV RNA was detected in the PBMCs of 29 patients (37.7%). After 6 months of the end of treatment, 15 (19.5%) of 77 ETR patients showed virological relapse, while 62 (80.5%) patients attained SVR. Relapse appeared significantly more often in patients with HCV RNA in their PBMCs at the ETR stage when compared to the patients who did not have the viral RNA (34.5% versus 10.4%, respectively; R2=6.67, P=.01; odds ratio [OR]: 1.3; 95% confidence interval [CI]=1.032–1.811). CONCLUSION Patients with HCV RNA in their PBMCs after attaining an ETR are more likely to show relapse as compared to patients who are negative for viral RNA in PBMCs at the ETR stage.

Preparation and characterization of a green nano-support for the covalent immobilization of glucoamylase from Neurospora sitophila

The preparation of green nano supports for the covalent immobilization of enzymes is of special interest both from the economic and environmental point of view. In this contribution, we report on the synthesis of phytochemicals coated silver nanoparticles, which were used as a novel green support for the covalent immobilization of glucoamylase isolated from Neurospora sitophila. The aqueous extract of Fagonia indica was used as a source of reducing and capping agents for the reduction of silver ions into silver nanoparticles. The prepared nanoparticles were characterized by various analytical techniques. UV-visible spectroscopy was used to detect the characteristic surface plasmon resonance bands (426, 438nm) of the silver nanoparticles. The biosynthesized silver nanoparticles were mostly spherical in shapes with an average particle size of 30-40nm (TEM and DLS measurements). X-ray diffraction and energy dispersive X-ray studies confirmed the face centered cubic crystalline form and elemental composition of the biogenic silver nanoparticles respectively. FTIR study revealed that plant polyphenolics and protein were mainly involved in the reduction and capping of silver ions. Glucoamylase from Neurospora sitophila was covalently immobilized to these nanoparticles via EDC (1-(3-(dimethylamino) propyl) 3-ethylcarbodiimidehydrochloride) coupling reaction. The immobilized enzyme exhibited higher pH and thermal stabilities as compared to the free enzyme. The kinetic constant (KM) value for the immobilized glucoamylase was higher (0.73mg/mL) than its free counterpart (0.44mg/mL), whereas the Vmax value was slightly higher for the immobilized glucoamylase. The findings of this study conclude that the newly developed green method for the synthesis of green nano-support is simple, cost effective and could be successfully used for the immobilization of various enzymes and other macromolecules.

Mesophilic Anaerobic Co-digestion of Cattle Manure with Malus domestica and Dalbergia sissoo during Biomethane Potential Assays

The burning of cattle manure for domestic use, and plant biomass left out in fields, is a common practice in South Asia, specifically Pakistan. According to the 2014 government of Pakistan (GOP) survey, Pakistan had 171 million head of cattle that would produce 345 billion kg of manure, which could easily be converted into 150 billion m3 of biogas. The focus of the present study was to evaluate the benefits from co-digestion of cattle manure (CM) with Dalbergia sissoo leaves (DSL) and Malus domestica leaves (MDL), with a focus on changes in the biodegradability, C/N ratio effect, and synergistic effect. The idea was to adjust the C/N ratio to increase biodegradability at mesophilic range to help the process to produce more methane than 100% manure-based digestion. First, the ideal pH and temperature conditions for mesophilic anaerobic digestion (AD) were optimized to carry out further co-digestion under the same conditions. The results of co-digestion revealed a 40% (251 NmL CH4/g VS) increase in methane yield by replacing 20% of volatile solid in CM-based AD reactors with MDL. This combination also presented a biodegradability of 59% and a synergistic effect (θ) value of 1.40, which corresponded to highly positive synergism reflecting the optimum growth conditions. The DSL/CM co-digestion also followed the same pattern, and the maximum methane yield of 229 NmL CH4/g VS was obtained using a 20/80 DSL/CM combination.

Use of Response Surface Methodology for the Biodegradation of Textile Industrial Effluents by Coniophora puteana IEBL-1

Currently almost all the textile industries use synthetic dyes as coloring agents because these gave more colors and are long lasting. Large proportion of these dyes released into water in the form of effluent.