Proteomic methods applied to the analysis of immobilized biocatalysts
Abstract Methods adapted from proteomics can directly characterize proteins present in immobilized biocatalysts. Complete hydrolysis followed by HPLC analysis of Tyr and Phe estimates total protein bound, and is preferable to conventional difference methods, as tested with subtilisin Carlsberg on si...
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crwiley:10.1002/bit.21074 2024-06-02T07:58:22+00:00 Proteomic methods applied to the analysis of immobilized biocatalysts Petry, Inga Ganesan, Ashok Pitt, Andrew Moore, Barry D. Halling, Peter J. 2006 http://dx.doi.org/10.1002/bit.21074 https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1002%2Fbit.21074 https://onlinelibrary.wiley.com/doi/pdf/10.1002/bit.21074 en eng Wiley http://onlinelibrary.wiley.com/termsAndConditions#vor Biotechnology and Bioengineering volume 95, issue 5, page 984-991 ISSN 0006-3592 1097-0290 journal-article 2006 crwiley https://doi.org/10.1002/bit.21074 2024-05-03T11:05:05Z Abstract Methods adapted from proteomics can directly characterize proteins present in immobilized biocatalysts. Complete hydrolysis followed by HPLC analysis of Tyr and Phe estimates total protein bound, and is preferable to conventional difference methods, as tested with subtilisin Carlsberg on silica. This new method shows that various treatments give quantitative desorption of proteins immobilized by adsorption. Intact desorbed proteins may be analyzed by electrospray mass spectrometry. The Candida antarctica lipase B from Novozyme 435 was shown to be heavily glycosylated, while the lipase from Lipozyme RM IM was a mixture of four N‐terminally truncated forms. Peptides from selective cleavage were analyzed by tandem mass spectrometry, leading to automatic identification of proteins present. A second major protein present in Lipozyme RM IM was thus found to be alpha‐amylase from Aspergillus oryzae . These methods should be valuable complements to activity measurements in understanding immobilized enzyme activity and stability. © 2006 Wiley Periodicals, Inc. Article in Journal/Newspaper Antarc* Antarctica Wiley Online Library Biotechnology and Bioengineering 95 5 984 991 |
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English |
description |
Abstract Methods adapted from proteomics can directly characterize proteins present in immobilized biocatalysts. Complete hydrolysis followed by HPLC analysis of Tyr and Phe estimates total protein bound, and is preferable to conventional difference methods, as tested with subtilisin Carlsberg on silica. This new method shows that various treatments give quantitative desorption of proteins immobilized by adsorption. Intact desorbed proteins may be analyzed by electrospray mass spectrometry. The Candida antarctica lipase B from Novozyme 435 was shown to be heavily glycosylated, while the lipase from Lipozyme RM IM was a mixture of four N‐terminally truncated forms. Peptides from selective cleavage were analyzed by tandem mass spectrometry, leading to automatic identification of proteins present. A second major protein present in Lipozyme RM IM was thus found to be alpha‐amylase from Aspergillus oryzae . These methods should be valuable complements to activity measurements in understanding immobilized enzyme activity and stability. © 2006 Wiley Periodicals, Inc. |
format |
Article in Journal/Newspaper |
author |
Petry, Inga Ganesan, Ashok Pitt, Andrew Moore, Barry D. Halling, Peter J. |
spellingShingle |
Petry, Inga Ganesan, Ashok Pitt, Andrew Moore, Barry D. Halling, Peter J. Proteomic methods applied to the analysis of immobilized biocatalysts |
author_facet |
Petry, Inga Ganesan, Ashok Pitt, Andrew Moore, Barry D. Halling, Peter J. |
author_sort |
Petry, Inga |
title |
Proteomic methods applied to the analysis of immobilized biocatalysts |
title_short |
Proteomic methods applied to the analysis of immobilized biocatalysts |
title_full |
Proteomic methods applied to the analysis of immobilized biocatalysts |
title_fullStr |
Proteomic methods applied to the analysis of immobilized biocatalysts |
title_full_unstemmed |
Proteomic methods applied to the analysis of immobilized biocatalysts |
title_sort |
proteomic methods applied to the analysis of immobilized biocatalysts |
publisher |
Wiley |
publishDate |
2006 |
url |
http://dx.doi.org/10.1002/bit.21074 https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1002%2Fbit.21074 https://onlinelibrary.wiley.com/doi/pdf/10.1002/bit.21074 |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_source |
Biotechnology and Bioengineering volume 95, issue 5, page 984-991 ISSN 0006-3592 1097-0290 |
op_rights |
http://onlinelibrary.wiley.com/termsAndConditions#vor |
op_doi |
https://doi.org/10.1002/bit.21074 |
container_title |
Biotechnology and Bioengineering |
container_volume |
95 |
container_issue |
5 |
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984 |
op_container_end_page |
991 |
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1800741692078292992 |