A 1.3 kb hyperthermophilic xyl-A gene encoding xylose isomerase from eubacterium Thermotoga naphthophila RKU-10 (TnapXI) was cloned and over-expressed in E. coli (BL21(DE3) to produce enzyme in mesophilic conditions that work at high temperature. The complete nucleotide sequence of the xyl-A gene was determined. Comparison of the nucleotide sequences with other xyl-A genes in the database showed that the xyl-A gene has 97% homology with that of the xyl-A gene from T. naphthophila available at NCBI. The inferred amino acid sequence showed that the enzyme was from class II of xylose isomerases. The TnapXI was concentrated by lyophilization and purified by heat treatment, fractional precipitation and UNOsphere Q anion-exchange column chromatography to homogeneity level. It was an acidic protein with theoretical isoelectric point (pI) 5.4 and theoretical molecular weight was calculated as 50.84 kDa. The apparent molecular mass (Mr) was estimated by SDS-PAGE to be 49.5 kDa. The active enzyme showed a clear zone on native-PAGE when stained with 2, 3, 5- triphenyltetrazolium chloride. The optimum temperature and pH for D-glucose to Dfructose isomerization were 98°C and 7.0, respectively. Xylose isomerase retains 85% of its activity at 50°C (t1/2 1732 min) for 4 h and 32.5% at 90°C (t1/2 58 min) for 2 h. It retains 90-95% of its activity at pH 6.5 to 7.5 for 30 min. The enzyme was highly activated (350%) with the addition of 0.5 mM Co2+ and to a lesser extent about 180 and 80% with the addition of 5 and 10 mM Mn2+ and Mg2+, respectively but it was inhibited (54-90%) in the presence of 0.5-10 mM Ca2+ with respect to apo-enzyme. t1/2 of TnapXI increased significantly by the addition of 1 mM Co2+ from 39.13% to 1466.67% as compared to apo-enzyme at temperature range 80-100°C. The enzyme showed a half life (t1/2) of 18 min for apo-enzyme (Kd 0.0385 min-1) and 65 min for holo-enzyme (Kd 0.0106 min-1) at 95°C. The catalytic affinities (Km) of the enzyme for xylose and glucose were 0.96 and 7.67 mM, respectively, while Vmax were 384 and 90 μmol/mg.min-1, respectively. The turn-over (kcat) rate was 5245 min−1 for D-xylose and 1229 min−1 for D-glucose. Catalytic efficiencies (kcat/Km) of enzyme for xylose and glucose were 5,463 and 160.2 min-1mM-1, respectively. The ionizable group of active site involved in controlling Vmax of the xxv enzyme, showed pKa1 and pKa2 as 6.0 and 7.6, respectively. The pKa1 and pKa2 were assigned to His-101 and His-271, respectively. Temperature quotient (Q10) was 2.05 while activation energy (Ea) was 82.25 kJ/mol. Thermodynamic parameters for Dglucose isomerization were ΔH* 79.19 kJmol-1, ΔG* -6.93×10−53 kJmol-1, ΔS* 215 Jmol- 1K-1, ΔG*E−S -14.9 kJmol-1 and ΔG*E−T -35.1 kJmol-1, at 368 K. The D values for apo and holo TnapXI were calculated as 1.776 and 2.336 min, respectively whereas the z values for apo and holo enzyme were calculated as 12.65 and 32.68°C, respectively at 95°C. The activation energy (Ea(d)) of isothermal irreversible deactivation at 95°C for apo and holo TnapXI were calculated as 209.5 and 770.1 kJ mol-1, respectively. The thermodynamic parameters i.e., ΔG*(d), ΔH*(d), and ΔS*(d) for deactivation of the apo-enzyme were 206.44 kJmol-1, 93.579 kJmol-1 and 0.306 Jmol-1K-1 and for the holoenzyme were 767.04 kJmol- 1, 104.56 kJmol-1 and 1.800 Jmol-1K-1, at 368 K. D-glucose isomerization product was also analyzed by thin layer chromatography (Rf 0.65). The enzyme was very stable at slightly acidic to neutral pH and have the greater tendency to resist the thermal unfolding at sufficiently high temperature and required only trace amount of Co2+ for its optimal activity and stability. Overall, 52.2% D-fructose was achieved by the isomerization of Dglucose using TnapXI. Thus, it has a great potential for industrial applications.
قاضی ظہورالحسن ناظمؔ سیوہاروی افسوس ہے گزشتہ مہینہ قاضی ظہور الحسن صاحب ناظمؔ سیو ہاری نے داعی اجل کولبیک کہا۔ مرحوم رشتہ میں راقم الحروف کے ماموں تھے۔بڑے ذہین، طباع،بذلہ سنج اورقادرالکلام شاعر تھے۔فن پربڑاعبور تھا۔تاریخ گوئی میں توشاید ہی اُن کاکوئی جواب ہو۔اس خاص کمال کی وجہ سے نظام حیدرآباددکن کے دامنِ دولت سے وابستہ ہوگئے اورعرصہ تک وظیفہ پاتے رہے۔غالباً۱۹۲۸ء میں جب ’’نظام‘‘ گورنمنٹ انڈیا سے برارکا معاملہ طے کرنے دلّی آئے تھے تومرحوم نے اس تقریب سے ایک عجیب وغریب قصیدہ لکھا تھا جس میں ۱۹ اشعارتھے اور ہرشعر سے سات طرح دلّی آنے کی تاریخ نکلتی تھی۔پھر لطف یہ تھا کہ ہرشعر کے پہلے حرف کوملائیے تواُسی بحر اوراُسی ردیف وقافیہ کاایک شعرہوجاتا تھا اوراس سے بھی سات طرح تاریخ برآمد ہوتی تھی۔نظام نے اس پر خوش ہوکر ان کے منصب میں اضافہ کردیا اوراب وہ مستقلاً حیدرآباد میں رہنے لگے تھے۔ علاوہ بریں اُن کو تصنیف وتالیف کا بھی بڑا متنوع ذوق تھا۔ تاریخ،فقہ،ادب و تنقید، لسانیات، شعروشاعری ان میں سے ہرموضوع پرانھوں نے کتابیں لکھی ہیں۔آخر میں اردو کی انسائیکلو پیڈیا لکھنی شروع کی تھی جوناتمام رہ گئی۔اخلاقی اعتبار سے خوش مزاج اورخندہ روبزرگ تھے۔لطائف وظرائف سینکڑوں کی تعداد میں یاد تھے اور انھیں موقع و محل کے لحاظ سے مزے لے لے کرسناتے تھے۔نماز باجماعت پڑھتے تھے اورادووظائف کاشغل بھی رکھتے تھے۔بزرگان دیوبند کے صحبت یافتہ اوران کے نام کے عاشق تھے۔دنیوی معاملات میں بھی بڑی سوجھ بوجھ رکھتے تھے۔اللھم اغفرلہ وارحمہ۔ [مئی۱۹۶۰ء]
The system of accountability is essential for the stability and development of any state. The department of accountability helps the Govt to evaluate the performance of its officer in order to block the way of corruption, dishonesty, and misuse of powers. Holy Prophet (ﷺ) introduced a new department for accountability and said that his pledge is the best time for accountability and named his era khayr-ul-quroon. These departments are legislators, judiciary, and administration. The institutions of accountability are summoned today as the Accountability Bureau. The aim of these institutions was to do work for the welfare of mankind and save humanity from the oppression of human beings. At an early age, these institutions were nominated as Dewan al-Ahtsab, Dewan- al-Mazloum. In Khilafat Rashida, many institutions were built on these concepts, and the Muslim ruler who came after them gave a complete system of Accountability. Later on, the names of these institutions were changed, but the aims of these institutions remained the same. This article explains the establishment of different types of accountability institutions in the era of the Holy Prophetﷺ and His companions and their function.
Stem end rot (SER) is a fungal disease complex that poses a major threat to the delivery of quality mangoes from Pakistan to export markets. The dynamics of this disease remain unclear, but must be elucidated so that effectiveness of disease management can be improved. The present study sought to describe the infection process and delineate a disease cycle of SER under Pakistani growing conditions. Culturing of symptomatic samples from traditional and good practices blocks revealed colonization of mango fruit by Lasiodiplodia theobromae, Phomopsis mangiferae, Colletotrichum gloeosporioides, Alternaria alternata and Botrytis cinerea, of which L. theobromae was the most prevalent. Fully mature but unripe mango fruits were collected from major mango growing belt of Punjab Province. The fruits were stored in cold storage (12oC; 21 days) and ambient storage (33oC; 9 days). After ripening, fruit under ambient and cold storage revealed that SER was the most prevalent disease. Pathogenicity assay revealed that all the artificially inoculated fungal pathogens were pathogenic. L. theobromae produced larger lesion diameter (74.6 mm) in ambient storage followed by C. gloeosporioides (54.6 mm). In cold storage, maximum lesion diameter (54.6 mm) was observed when spore suspension of L. theobromae was inoculated followed by C. gloeosporioides (37.6 mm). In another study, the relationship between lengths of fruit pedicle with SER disease development was investigated. It was found that retaining a portion of the pedicel on harvested fruit reduced development of SER during the course of ripening, in comparison to fruit from which the pedicel was removed. Histopathology of pedicel attached with mango fruit indicated an extensive distribution of L. theobromae infecting xylem vessels and other tissues. In subsequent research, the efficacy of fungicides - Cabrio Top®, Nativo®, Scholar®, Tecto®, Amistar®, and Sportak®, and plant extracts - ( Peganum harmala, Syzygium aromaticum, Cichorium intybus, Moringa oleifera, Cinnamomum zeylanicum and Coriandrum sativum) at several concentrations was assessed against SER pathogens in vitro and in vivo. Results indicated that Nativo and Cabrio Top at the concentration of 200 and 250 μg/mL, and M. oliefera, S. aromaticum and C. zeylanicum at the concentration of 25 and 50 μg/mL, provided statistically significant suppression of mycelial growth of tested pathogens and SER development.