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Home > Characterization of Indigenous Bacterial Strains Involved in the Production of Endo-Polygalacturonases

Characterization of Indigenous Bacterial Strains Involved in the Production of Endo-Polygalacturonases

Thesis Info

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Author

Hadri, Saqib Hussain

Program

PhD

Institute

Pir Mehr Ali Shah Arid Agriculture University

City

Rawalpindi

Province

Punjab

Country

Pakistan

Thesis Completing Year

2019

Thesis Completion Status

Completed

Subject

Biochemistry

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/10800/1/Saqib%20Hussain%20Hadri_Biochem_2019_PMAS_PRR.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676725688814

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The current study was carried out to isolate indigenous bacteria for the production of endo-PG. Bacteria were isolated, purified and screened initially on the basis of morphology, pigment production, margins etc. Ten isolates were selected after the initial screening, which were identified from their 16S rRNA gene sequence. The ten isolates were named IEB-1 to IEB-10. DNA was isolated from the bacteria and 16S rRNA gene was amplified by PCR using universal primers like 27F and 1492R, which were modified for cloning in pTXB1. The PCR product was cloned in pTXB1 and the plasmids were sent for sequencing from University of Rochester Center of Genomics. The 16S rRNA gene sequence was blasted on NCBI nucleotide database. The isolates were identified as Bacillus subtilis, Bacillus pumilius, Exiguobacterium acetylicum, Bacillus pumilius, Staphylococcus hominis, Bacillus licheniformis, Serratia rubidaea respectively from IEB-1 to IEB-9. Nucleotide blast of 16S rRNA gene sequence of IEB-10 showed less than 95% resemblances with many of the species. The selected ten bacterial isolates were then screened for the production of the endopolygalacturonase through different steps. Three isolates showed endo-PG activity. Two isolates were identified as Bacillus pumilus and one was identified as Bacillus licheniformis. The later one has already been reported to be endo-PG positive, but Bacillus pumilus has only been reported as pectinase positive but not as endo-PG positive. Gene sequence of endo-PG from B. pumilus has also not been reported earlier. Primers were designed on the basis of the glycoside hydrolase gene from B. pumilus. The PCR product of glycoside hydrolase was cloned in the double digested pTXB1 which was transformed in E. coliBL21competent cells. E. coli colonies containing the cloned “glycoside hydrolase” gene gave a positive pectinase plate screening assay. B. pumilus was used for the production and characterization of endo-PG. Citrus peals, reddish peals and apple peals were used as the only source of nutrients during liquid state fermentation for B. pumilus for the production of endo-PG to make its production cheaper. Response surface methodology (RSM) under central composite design (CCD) was used for the optimization of various parameters, for the production of the endo-PG, including temperature, pH, inoculum size, inoculum age, fermentation period and substrate water ratio. Three dimensional graphs and contour plots were prepared using JMP-12 software. The optimized results showed the best production of the endo-PG at 39 oC, pH of 7.03, 52 hours of fermentation period, 3.16 % substrate water ratio, 6.5mL of inoculum size having 16 hours of inoculum age. Different carbon, nitrogen sources, surfactants and mediators were used to check their effect on the endo-PG production. All of the carbon sources and some of the nitrogen sources gave negative effect on the production of endo-PG. Ca+2 and Zn+2 showed very negative effect on the endo-PG activity, while Mg+2 had not particular effect at its low concentrations, but higher concentrations of Mg+2 also gave negative effect on the endo-PG activity. The enzyme showed maximum activity at pH 7 and 40 oC. A concentration of 70% was found to be the optimum for the partial purification of the endo-PG. The dialyzed partially purified extract was applied to column chromatography for the further purification. The elution showing a positive endo-PG assay was run on the SDS-PAGE to check the purity and molecular weight of the protein. The stained SDS-PAGE gel showed a single band at 45kDa approximately.
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بنیادی ماخذات

کتابیات

بنیادی ماخذات

ٔ٭          ناطق علی اکبر، بے یقین بستیوں میں، سٹی پریس بک شاپ، کراچی، 2010ء

٭             ناطق علی اکبر، در عدالت علے، عکس پبلیکشنز ، لاہور،2020ء

٭             ناطق علی اکبر، ریشم بننا کھیل نہیں، سانجھ پبلیکیشنز لاہور، 2019ء

٭             ناطق علی اکبر، سربستیوں کے غزال، سانجھ پبلیکشنز ،لاہور، 2018ء

٭             ناطق علی اکبر، سرمنڈل کا راجہ، سانجھ پبلیکشنز لاہور، 2016ء

٭             ناطق علی اکبر، سفیر لیلیٰ کلیات، بک کارنر، جہلم، 2022ء

٭             ناطق علی اکبر، شاہ محمد کا ٹانگہ،،سانجھ پبلیکشنز ، لاہور،2018ء

٭             ناطق علی اکبر، فقیر بستی میں تھا،عکس پبلیکشنز، لاہور،2019ء

٭             ناطق علی اکبر، قائم دین افسانے، سانجھ پبلیکیشنز ، لاہور،2018ء

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٭             ناطق علی اکبر،ھیت شعر، اسوۃکالج، اسلام آباد، 2016ء

ثانوی ماخذات

٭             احمد، عزیز، آگ، شاہراہ ،دہلی،2000ء

٭             اسلم آزاد، ڈاکٹر،اردو ناول آزادی کے بعد، کمانت پرکاش نزایا، بہرام دہلی، 1990ء

٭             اشفاق انیس ، دکھیارے،شہزاد،D-155، بلاک نمبر 5 گلشن اقبال، کراچی 2014ء

٭             تارڑ مستنصر حسین، خس وخاشاک زمانے، سنگ میل پبلیکشنز لاہور، 2010ء

٭             جاوید اختر ڈاکٹر، اردو کی ناول نگار خواتین، سنگ میل پبلیکیشنز،لاہور،1997ء

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Ibn e Khaldun (1406 C.E.) has been an imminent scholar and well known for his work in the study of civilization. His vision regarding Civilization holds the significant place according to the philosophers of history. Arnold J. Toynbee (1975 C.E.) is one such prominent thinker who not only applauded the thoughts of Ibn e Khaldun but was influenced by Ibn e Khaldun’s views as it can be seen in Toynbee’s book: “A Study of History”. As a philosopher of history, he has much contribution in the field; He interlinks History with civilization. He presented a thoughtful book surrounding his civilizational vision; which explains the causes of world’s ups and downs. Although he presented a quality research about the division of the civilizations in the light of religion, many aspects of his work need to be reassessed. As per his understanding of world civilizations, he represents twenty-one civilizations, but with the passage of time, the number reduces and now only five are left in the contemporary epoch. According to him, religion has played significant role in the rise and fall of civilizations through their various stages. This research will highlight his thought about Islam through a comparison between Toynbee and Ibn e Khaldun’s Islamic civilizational thoughts. The study will also mention several problems in his approach to the Islamic Civilization. Furthermore, along with due importance of both scholars in the subject of history, their authoritative status will be stated. This research aims to discuss some misconceptions of the West that are based on Toynbee's understating of Islamic civilization and history. And, consequently, it intends to improve relations between people of the west and east.

Sino-Indian Quest for Hegemony in South Asia: Implications for Pakistan

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