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Home > Production, Purification and Characterization of Carboxymethyl Cellulase for Food Applications

Production, Purification and Characterization of Carboxymethyl Cellulase for Food Applications

Thesis Info

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Author

Nadeem, Muhammad Tahir

Program

PhD

Institute

University of Agriculture

City

Faisalabad

Province

Punjab

Country

Pakistan

Thesis Completing Year

2009

Thesis Completion Status

Completed

Subject

Applied Sciences

Language

English

Link

http://prr.hec.gov.pk/jspui/handle/123456789/178

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676727031542

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Agricultural wastes/byproducts have great potential for the production of value added products with special reference to enzymes. Two indigenous carbon sources namely, wheat bran and sugar cane bagasse were used as substrates for carboxymethyl cellulase (CMCase) synthesis by Aaspergillu niger through submerged fermentation. The fungus produced highest CMCase activities at 3% level of wheat bran using 30°C and pH 5 over 96 hrs of incubation. The enzyme was purified for its further use in food processing. During the purification process, a 3.05 fold purification of enzyme was observed after gel filtration. Characterization of purified enzyme was carried out to find out the most suitable conditions for the CMCase performance; the best enzyme activity was observed at 50°C and pH 4.5. The enzyme was quite stable and retained 37% of its maximum activity at 90 ° C. The SDS-PAGE of the purified enzyme revealed its molecular weight as 47.1 kDa. For efficacy purpose, the enzyme added feeds were given to the broiler chicks in order to investigate their effects on body performance, feed efficiency and for safety concerns. The enzymatic treatment exhibited positive effects on body weight, carcass weight and chest muscle weight. Non-significant variations in the serum biochemistry and hematological parameters were observed. After safety evaluation, the enzyme was employed at various levels in bread preparation. Different treatments exhibited significant variations in rheological properties of the dough. The CMCase application in the bread making process resulted in increased volume (15.45%) up to a level of 250 IU/100g flour (T 2 ) however, excessive dose showed a decreasing trend for this parameter. The treatment T 2 produced bread with specific volume 3.99 cm 3 /g in contrast to that of T 0 i.e. 3.37 cm 3 /g. The external and internal characteristics of bread were also improved by the enzyme addition. Later, the enzyme application in citrus juice extraction also showed beneficial effects. Although the enzymatic treatment had non-significant effect on TSS, density and pH of pulp and juice however, a momentous reduction in pulp viscosity i.e. 74.79% was observed in T 6 (CMCase and pectinase 50 IU/100g each). Moreover, the treatment T 6 gave more clarified juice with 95.70% transmittance than that of control (78.00%); also resulted in 20.00% increase in the juice yield with 90.00% juice recovery. The sensory evaluation of the enzyme treated juice reflected improved taste, flavor and sweetness however; caused slight color deterioration of the resulting juice. The purified enzyme was also employed in carotenoid extraction from citrus peel and the maximum yield (8.60±0.94 mg/100g peel) was calculated in case of T 4 (250 IU of each enzyme/100g peel). The pigment showed more stability in dark as compared to light at 30°C. Furthermore, higher pigment loss was observed at 45°C than that at refrigeration temperature (4°C). It was also observed that freeze drying of the pigment enhanced its stability. It is concluded from the present investigation that wheat bran has a potential for CMCase production that can efficiently be used in bread production, juice processing and carotenoid extraction.
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خود کو دیکھ رہا ہوں کیا ہے

خُود کو دیکھ رہا ہوں کیا ہے
وہ کیوں مجھ کو چھُوڑ گیا ہے

پھر دل غم سے نڈھال ہوا ہے
ہجر تعلق ، ٹوٹ چکا ہے

یاروں سے کیا شِکوہ کرنا
بس یوں ہی دل بھر آیا ہے

اِس دل کی نگری کا راجا
پتھّر ہے یا پتھّر کا ہے

اَب بُلبُل کو شب ہو جائے
کب کوئی جُگنو آتا ہے

کلی جو کھل کر پھول بنی تھی
پھول کسی نے توڑ لیا ہے

سارے پتّے زرد ہوئے ہیں
پیڑ کو کیا غم ہو سکتا ہے!

لوگ پوچھتے ہیں کیا غم ہے
رنگ ترا بھی ناصرؔ سا ہے

ساری دنیا پاس ہے صادقؔ
تو پھر بھی کتنا تنہا ہے

Al-Sukākī’s Classification of Metaphor and Qurānic Discourse

The present study is divided into two main sections; the first section will give a general overview about the figurative language and more focus on metaphor (istiᶜārah in Arabic) because the metaphor is considered as one of the most literary devices and the main category of the figurative language. So in this study has given various definitions of figurative language and metaphor according to Muslims and Non-Muslims linguists and along with this explained Al-sukākī’s classification of metaphor which is little close to Al-Jurjānī’s classification of metaphor and view respectably among Muslims and Non-Muslims linguists. The second section of this study deals with metaphors given in Holy Qur'ān, which are denoted according to Al-sukākī’s classification in this respect. In this reference the verses are presented with detailed tafsīrī literature so the reader could well comprehend the purposes and the classical aspect of metaphors in text and also could evaluate linguistic architecture of  Holy Qur'ān.

Synthesis and Bioactivity Evaluation of Amoxicillin Derivatives

The work presented in this thesis consists of “synthesis and bioactivity evaluation of amoxicillin derivatives” which includes the synthesis, optimization, applications and bioactivity evaluation of new metallic derivatives (silver and gold nanoparticles) of amoxicillin and some other beta lactam antibiotics. For a wide spread resistive strains of pathogenic bacteria, there is a need for some new antibacterial agents to treat the patients, infected with such resistive pathogenic bacteria. Here we synthesized new gold and silver derivatives of amoxicillin and some other beta lactam antibiotics like cefuraxime, cephradine, ceftriaxone etc belonging to beta lactam antibiotics family; some of them were penicillins while others are cephalosporins. We synthesized metallic derivatives of amoxicillin in the form of gold and silver nanoparticles and their nano-alloys using NaBH4 and TMA as reducing agents. Reducing agents of different nature (organic and inorganic), NaBH4 and trimethylamine were used for the synthesis of gold and silver nanoparticles to study their effect on sizes and shapes of nanoparticles as well as to avoid the sodium salt formation of amoxicillin and other beta-lactam antibiotics using TMA. It was found that nanoparticles synthesized with NaBH4 were of small size compared to nanoparticles synthesized with TMA. Gold nanoparticles stabilized with amoxicillin were prepared using NaBH4 and TMA as reducing agents. The applications for metallic sensing like Cu, Fe, Co, Ni, Pb, Cd, and Hg etc were studied. Gold nanoparticles stabilized with amoxicillin were fluorescent and were used for the detection of Cu2+ions in water solution. Amoxicillin and gold nanoparticles (AuNPs) stabilized with amoxicillin were screened for enzymes inhibition studies as well antiglycation and results were compared. These AuNPs showed very nice inhibition for urease, carbonic anhydrase and xanthine oxidase enzymes and were found very potent inhibitor for xanthine oxidase enzyme with inhibition of 90% and IC50 value of 2.0±0.01. Amoxicillin showed antiglycation activity while AuNPs were inactive. xv Silver nanoparticles stabilized with amoxicillin were also prepared using NaBH4 and TMA as reducing agents. AgNPs were screened for enzymes inhibition studies and antiglycation activity and results were compared with amoxicillin and it was found that AgNPs were active against urease and xanthine oxidase enzymes while inactive against carbonic anhydrase and no antiglycation activity. Nano-alloys of AgNPs and AuNPs stabilized with amoxicillin were also prepared. These were also screened for enzymes inhibition and antiglycation activity but were inactive. Gold and silver nanoparticles using other stabilizing agents like cefuroxime, cephradine, ceftriaxone and cefixime were also synthesized. Their nano-alloys were also prepared stabilized with the above mentioned antibiotics except cefixime. Their applications for metals sensing like Cu, Fe, Co, Ni, Pb, Cd, and Hg etc. were studied. The metallic sensing application of cefixime was investigated and was found that AgNPs stabilized with cefixime detected Ni ion in water solution while its AuNPs detected Cu ion in water solutions. Cephradine stabilized gold nanoparticles detected Hg and Cd ions in water solution upto micro molar concentration. The comparative bioassays of gold and silver nanoparticles stabilized with cefuroxime, cephradine, ceftriaxone and cefixime as well as their parent compounds were also studied and compared. Cefuroxime and their gold and silver nanoparticles showed best results in case of urease inhibitions. Cefuroxime was also found to be active by inhibiting urease enzymes along with its antibiotic activities. AuNPs stabilized with cefuroxime were also active against carbonic anhydrase enzymes. While its silver and nan-alloys were active by inhibiting in Vitroα-Chymotrypsin enzymes. Cephradine stabilized silver nanoparticles and nano-alloys were active for xanthine oxidase enzyme with an inhibition of 87.9% and 85.9% with IC50 value 6.9±1.5 μg/mL and IC50 value 6.76±0.07 μg/mL respectively. AuNPs and parent compound were inactive against XO enzymes inhibition. AuNPs, AgNPs, nano-alloys and parent compound cephradine were active for urease enzymes inhibition and inactive for antiglycation activity except the parent compound. AuNPs and AgNPs were active for carbonic anhydrase inhibition while Nano-alloys and parent compound were inactive. All were inactive for In Vitroα-Chymotrypsin enzyme inhibition. Ceftriaxone caped gold and silver nanoparticles were active for urease inhibition except its nano- alloy while only AgNPs were active for xanthine oxidase enzyme inhibition and rest of members were inactive. All were inactive for the remaining enzymes as well as antiglycation activity. Cefixime is active against xanthine oxidase enzyme inhibition and antiglycation activity while gold and silver nanoparticles were active for urease enzyme inhibition only.