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Utilization of Industrial Wastes for the Production of Amylase from Thermophillic Actinomycetes

Thesis Info

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Author

Khan, Muhammad Ayub

Program

PhD

Institute

Hazara University

City

Mansehra

Province

KPK

Country

Pakistan

Thesis Completing Year

2012

Thesis Completion Status

Completed

Subject

Microbiology

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/9616/1/1272.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676727652661

Similar


Actinomycetes are a facultative thermophilic group of gram-positive bacteria which produce antibiotics and a range of hydrolytic enzymes including amylases, proteases and lipases. Some Actinomycetes are pathogenic but the majority is saprophytic inhabiting soil, water plants and other habitats, where they play an important role in soil structure and composition. In the present study, bacterial strains from three soil samples obtained from a hot water stream were screened for their potential to produce α -amylase. Out of twenty isolated strains, four were selected due to their amylase producing ability as determined through production of a zone of hydrolysis. These strains were identified by morphology, biochemical tests and PCR amplification. Bacterial strain W1 giving a 3.8 mm zone of hydrolysis was identified to be Actinomycete and N3, U1, and U4 giving 3.6, 3.8 and 3.6 mm zones of hydrolysis, respectively were identified as Bacilli. Growth of Actinomycete and amylase production was optimized on four different nutrient media. Highest growth and amylase activity( 3.882 OD and 132.3 AU, respectively after 72 hours at 60oC) were obtained by using medium-4. Conditions for the maximum production of amylase in submerged culture fermentation were optimized using medium-4. It was found that by using 5% inoculum of 30 hours age, amylase activity of 140.8 AU was obtained. Optimization of agitation speed (150 rpm), temperature (60 oC), pH (7.5), starch (2.5 %) and glucose (3%) increased activity from 140.8 AU to 162.4 AU. Growth and amylase production were then optimized by using agro-industrial waste as sole source of carbon and nitrogen in the modified AGS medium. Nine agro-industrial wastes namely soybean meal, wheat bran, peanut meal, mustard meal, sunflower meal, linseed meal, cottonseed meal, corn meal and molasses were used individually and in combinations to optimize amylase production. Soybean meal and wheat bran were found to be more effective nutrient sources with amylolytic activities of 176.2 AU and 171.6 AU respectively. Soybean meal in combination with molasses at 2:1 ratio were found to bethe best carbon and nitrogen sources giving amylolytic activity of 230.6 AU. In solid state fermentation, maximum amylolytic activity( 186.2 AU) was obtained by using wheat bran and molasses in 2:1 ratio at 55oC after 72 hours of incubation. The enzyme was purified 24-fold by acetone precipitation and gel filtration. Single peak during gel filtration indicated presence of only one type of alpha-amylase. Crude and partially purified enzyme find vital application in desizing of cotton cloth. Desizing of cotton pieces 15x15 cm by acid hydrolysis (0.1 M HCl) resulted in 0.7725g removal of starch. This value served as standard for 100% starch removal. Crude enzyme gave 98.39% removal after 90 minutes of soaking at pH 7.0. However, 98.92 % removal was achieved with partially purified enzyme after 60 minutes of soaking. Optimization of temperature and pH revealed that 100 % desizing was obtained at pH 6.0 when incubated at 60oC for 60 minutes. Studies proved to be successful in utilizing agro-industrial waste for producing thermostable alpha-amylase from Thermoactenomyces sacchari and its application as a desizer. The process is recommended for large production of thermostable alpha-amylase in the future.
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۔غزل

غزل۔۔۔محمد ایوب صابر

بجھ گیا ہے دل سے شعلہ عظمتِ پرواز کا
بے پروں کے جھنڈ دیکھے  ہیں زمیں پر رینگتے
صورتِ شاہیں کرے تسخیر جو دلکش فضا
وہ پرندہ شوق سے اڑتا پھرے شام و سحر
عمر بھر اس نے تھکن سے واسطہ رکھا نہیں
آسماں کی سمت اڑنے میں مری توقیر ہے
جو قفس میں زندہ رہنے  کے لئے تیار  ہیں
ہمتِ پرواز میرے خوں میں ہے محوِ سفر
سربلندی کی طرف صابر اسے مائل کرو

 

پڑ گیا ہے ماند  جذبہ عظمتِ پرواز کا
سن رہے ہیں صرف قصہ عظمتِ پرواز کا
اس کے سر پر باندھ سہرا عظمتِ پرواز کا
پی لیا ہے جس نے دریا عظمتِ پرواز کا
چکھ لیا تھا جس نے خوشہ عظمتِ پرواز کا
مل گیا ہے مجھ کو زینہ  عظمتِ پرواز کا
پڑھ رہے ہیں مل کے نوحہ عظمتِ پرواز کا
سوچ میں باقی ہے نقشہ عظمتِ پرواز کا
پوچھتا ہے جو بھی  رتبہ عظمتِ پرواز کا

Multiple Intelligences: Learners VS Teachers

The study investigated the relationship of the multiple intelligences of the Bachelor of Secondary Education students and their teachers in their major subjects. Four hundred eighty-five (485) BSED students and twenty-two (22) teachers in their respective major subjects participated. The result demonstrates statistically significant in the multiple intelligences of the Bachelors of Secondary Education Major in Technology and Livelihood Education and Music, Arts, Physical Education and Health and their teachers in their respective major subjects. However, result also demonstrates no significance in the multiple intelligences of the Bachelors of Secondary Education Major in Filipino, English, and Mathematics and their teachers in their respective major subjects. The study shows that the dominant intelligences of the BSED students and their teachers in their major subjects are the interpersonal, intrapersonal, and their suited intelligences for their major subjects. The result evidently showed that the BSED students and their major teachers are people and self smart. This only shows that as a teacher, one should know how to socialize appropriately with others and have a deeper understanding with themselves. It also showed that the teachers are really smarter than their students in their major field of specialization. Educators must also consider the multiple intelligences of their students to fully develop their learning capabilities.

Synthesis and Structural Characterization of Mixed Ligand Complexes of Copper Ii With Diamines and Carboxylates

This research work presents one pot synthesis of ternary copper(II) carboxylates of N,Nchelating diamine ligands. The carboxylate ligands used were sodium salts of benzoic acid, 2-chlorobenzoic acid, cinnamic acid, succinic acid, phthalic acid, terephthal ic acid, 4- aminobenzoic acid, 3-aminobenzoic acid, mefenamic acid, acetyl salicylic acid, and tartaric acid. The N,N-chelating diamines utilized include N,N,N′,N′-tetramethylethylenediamine (tmen), while some complexes of carboxylates with ethylenediamine (en), 1,10– phenanthroline (phen) and 2,2′–bipyridine (bipy) have also been prepared. The structural aspects and geometrical assignments related to the synthesized complexes have been investigated with the help of analytical techniques like FT-IR, UV-Visible spectroscopy, thermal studies (TGA) and single crystal X-ray diffraction analysis. The investigated ternary copper(II) complexes involving N,N,N′,N′-tetramethylethylenediamine (tmen) include [Cu(tmen)(BA)2(H2O)2], (1a), [Cu(tmen)(salH)2(H2O)] (2a), {[Cu(tmen)(mef)2] (3a), [Cu(tmen)(pABA)2]. 1/2 MeOH) (4a), [Cu(tmen)(o-ClBA)2] (5a), [Cu(tmen)(cinn)2]. H2O (6a), [Cu(tmen)(phtH)2] (7a), [Cu(tmen)(tpht)(H2O)2]n (8a), {[Cu(tmen)(succin)]n.4H2O} (9a), {[Cu(tmen)(tart)]·2H2O}n (10a). Single crystal analyses of the prepared complexes have revealed that most of the Cu(II)-N,N,N′,N′-tetramethylethylenediamine adducts with the carboxylate ligands are mononuclear, in which N,N,N′,N′-tetramethylethylenediamine is coordinated to Cu(II) in an invariably chelating bidentate mode. In these complexes, the carboxylate moiety belonging to a carboxylate ligand is coordinated to the central Cu(II) ion, either in a monodentate (1a, 2a, 4a, 7a), or bidentate (3a, 5a, 6a) fashion. These mononuclear complexes can be; four-coordinate (4a), with a square planar environment, five-coordinate (2a, 7a),with a square pyramidal geometry, or six-coordinate (1a, 3a, 5a, 6a ) with an octahedral coordination geometry. Three complexes of Cu-tmen-carboxylato series are polynuclear in nature (8a, 9a, 10a) and adopt an octahedral coordination environment. The carboxylate functionality varies in coordination modes, from bis-monodentate bridging (8a) to chelating bridging (9a-10a). Another mixed ligand copper(II) complex incorporating ethylenediamine and salicylate [Cu(en)(salH)Cl]n (where en= ethylenediamine, salH1- = (salicylate1-) (11a) has also been synthesized. The complex [Cu(en)(salH)Cl]n (11a) is found to be unprecedented because of ix the presence of [Cu-Cl]n back-bone formed by central Cu(II) ion and bridging Cl atoms, also it had both ethylenediamine and salicylic acid as a part of the inner coordination sphere, while in most of the known examples, carboxylates usually are found lying uncoordinated in the outer sphere. Two ternary copper(II) carboxylate complexes, containing 2,2′-bipyridine (bipy = C10H8N2) having the formulae [Cu(bipy)(cinn)2(H2O)] (1b) [Cu2(bipy)2(pABA)3(pABAH)]. Cl. 3H2O (2b) {(where cinn1- = cinnamate (C9H7O2 1-) anion, pABA1- = p-amino benzoate (C7H6NO2 1-) anion, and pABAH = p-amino benzoic acid (C7H7NO2)} have been prepared and characterized. The mononuclear ternary Cu(II) complex incorporating 2,2′–bipyridine and cinnamate as shown by single crystal X-ray analyses is found to be square pyramidal, formed by the coordination of bidentate 2,2′–bipyridine, and two monodentate carboxylate groups from two cinnamates, while the apical position is occupied by an aqua-O atom. The second dinuclear mixed ligand Cu(II) complex of 2,2′–bipyridine and p-aminobenzoate (1b) is also found to be unique. It has two copper(II) centers in square pyramidal environments, which are interlinked by two bridging p-aminobenzoates and by two 2,2′–bipyridine ligands in a chelating mode. One remaining p-aminobenzoate is attached through its carboxylato-O atom in a traditional monodentate mode, while the other pABAH is attached to copper(II) through its N atom. Two novel mixed ligand copper(II)-phen based carboxylate complexes represented as [Cu(phen)(benzoate)2] 1c, and [Cu(phen)(m-amb)Cl·½H2O] 2c (where phen = 1,10- phenanthroline, BA1- = benzoate, m-ABA1- = m-aminobenzoate) have been synthesized and characterized. The geometry and structure of the mononuclear ternary Cu(II) complex incorporating 1,10–phenanthroline and m-aminobenzoate, as confirmed through single crystal X-ray analyses is found to be square pyramidal, formed by the coordination of chelating 1,10–phenanthroline, a chelating m-aminobenzoate, while the apical position is occupied by a Cl atom. The second monomeric complex [Cu(phen)(benzoate)2] was square planar, with one bidentate phen and two monodentate benzoates. Antimicrobial studies of complexes have also been performed. Some of these copper(II) complexes are found to be biologically active against bacteria.