Partial purification of cyclodextrin glucanotransferase from alkalophilic Bacillus sp. TS1-1

The cyclodextrin glucanotransferase (CGTase; EC 2.4.1.19) from Bacillus sp. TS1-1 has been partially purified from crude culture by centrifugation and cross-flow filtration. Initial 72.96 U/ml of CGTase was detected in the culture after 24 hours of incubation. The crude supernatant obtained after ce...

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Main Author: Mohd Azim, Abdul Rahman
Format: Undergraduates Project Papers
Language:English
Published: 2008
Subjects:
Online Access:http://umpir.ump.edu.my/id/eprint/792/1/Partial%20purification%20of%20cyclodextrin%20glucanotransferase%20from%20alkalophilic%20Bacillus%20sp.%20TS1-1.pdf
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author Mohd Azim, Abdul Rahman
author_facet Mohd Azim, Abdul Rahman
author_sort Mohd Azim, Abdul Rahman
collection UMP
description The cyclodextrin glucanotransferase (CGTase; EC 2.4.1.19) from Bacillus sp. TS1-1 has been partially purified from crude culture by centrifugation and cross-flow filtration. Initial 72.96 U/ml of CGTase was detected in the culture after 24 hours of incubation. The crude supernatant obtained after centrifugation for 5000 rpm, 5 minutes and 4°C was subsequently filtered at 15°C through cross-flow filtration using Kvick Lab cross-flow system. Two cassettes were used with molecular weight cut off of 50K and 10K. The retentate from 50K cassette was further filtered through 10K cassette. Each permeate and retentate from each cassette were tested for CGTase activity. The highest CGTase activity was detected at retentate of 10K cassette which suggested an initial size of CGTase between 10 kDa and 50 kDa. The crude and partially purified enzyme was then subjected to electrophoresis (SDS-PAGE). It was clear that a large amount of proteins had been removed by cross-flow filtration. Based on this work, the crude enzyme was purified 5.57 fold using cross-flow filtration.
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spelling UMPir7922023-10-25T07:05:22Z http://umpir.ump.edu.my/id/eprint/792/ Partial purification of cyclodextrin glucanotransferase from alkalophilic Bacillus sp. TS1-1 Mohd Azim, Abdul Rahman TP Chemical technology The cyclodextrin glucanotransferase (CGTase; EC 2.4.1.19) from Bacillus sp. TS1-1 has been partially purified from crude culture by centrifugation and cross-flow filtration. Initial 72.96 U/ml of CGTase was detected in the culture after 24 hours of incubation. The crude supernatant obtained after centrifugation for 5000 rpm, 5 minutes and 4°C was subsequently filtered at 15°C through cross-flow filtration using Kvick Lab cross-flow system. Two cassettes were used with molecular weight cut off of 50K and 10K. The retentate from 50K cassette was further filtered through 10K cassette. Each permeate and retentate from each cassette were tested for CGTase activity. The highest CGTase activity was detected at retentate of 10K cassette which suggested an initial size of CGTase between 10 kDa and 50 kDa. The crude and partially purified enzyme was then subjected to electrophoresis (SDS-PAGE). It was clear that a large amount of proteins had been removed by cross-flow filtration. Based on this work, the crude enzyme was purified 5.57 fold using cross-flow filtration. 2008-05 Undergraduates Project Papers NonPeerReviewed pdf en http://umpir.ump.edu.my/id/eprint/792/1/Partial%20purification%20of%20cyclodextrin%20glucanotransferase%20from%20alkalophilic%20Bacillus%20sp.%20TS1-1.pdf Mohd Azim, Abdul Rahman (2008) Partial purification of cyclodextrin glucanotransferase from alkalophilic Bacillus sp. TS1-1. Faculty of Chemical & Natural Resources Engineering, Universiti Malaysia Pahang.
spellingShingle TP Chemical technology
Mohd Azim, Abdul Rahman
Partial purification of cyclodextrin glucanotransferase from alkalophilic Bacillus sp. TS1-1
title Partial purification of cyclodextrin glucanotransferase from alkalophilic Bacillus sp. TS1-1
title_full Partial purification of cyclodextrin glucanotransferase from alkalophilic Bacillus sp. TS1-1
title_fullStr Partial purification of cyclodextrin glucanotransferase from alkalophilic Bacillus sp. TS1-1
title_full_unstemmed Partial purification of cyclodextrin glucanotransferase from alkalophilic Bacillus sp. TS1-1
title_short Partial purification of cyclodextrin glucanotransferase from alkalophilic Bacillus sp. TS1-1
title_sort partial purification of cyclodextrin glucanotransferase from alkalophilic bacillus sp ts1 1
topic TP Chemical technology
url http://umpir.ump.edu.my/id/eprint/792/1/Partial%20purification%20of%20cyclodextrin%20glucanotransferase%20from%20alkalophilic%20Bacillus%20sp.%20TS1-1.pdf
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