Multienzyme catalysis in microfluidic biochips
Abstract The attachment of enzymes to glass microfluidic channels has been achieved using a highly reactive poly(maleic anhydride‐ alt ‐α‐olefin) (PMA)‐based coating that is supplied to the microchannel in a toluene solution. The PMA reacts with 3‐aminopropyltriethoxysilane groups linked to the glas...
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crwiley:10.1002/bit.10642 2024-06-02T07:58:21+00:00 Multienzyme catalysis in microfluidic biochips Lee, Moo‐Yeal Srinivasan, Aravind Ku, Bosung Dordick, Jonathan S. 2003 http://dx.doi.org/10.1002/bit.10642 https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1002%2Fbit.10642 https://onlinelibrary.wiley.com/doi/pdf/10.1002/bit.10642 en eng Wiley http://onlinelibrary.wiley.com/termsAndConditions#vor Biotechnology and Bioengineering volume 83, issue 1, page 20-28 ISSN 0006-3592 1097-0290 journal-article 2003 crwiley https://doi.org/10.1002/bit.10642 2024-05-03T11:44:17Z Abstract The attachment of enzymes to glass microfluidic channels has been achieved using a highly reactive poly(maleic anhydride‐ alt ‐α‐olefin) (PMA)‐based coating that is supplied to the microchannel in a toluene solution. The PMA reacts with 3‐aminopropyltriethoxysilane groups linked to the glass surface to form a matrix that enables additional maleic anhydride groups to react with free amino groups on enzymes to give a mixed covalent–noncovalent immobilization support. Using a simple T‐channel microfluidic design, with reaction channel dimensions of 200 μm wide (at the center), 15 μm deep, and 30 mm long giving a reaction volume of 90 nL, soybean peroxidase (SBP) was attached at an amount up to 0.6 μg/channel. SBP‐catalyzed oxidation of p ‐cresol was performed in aqueous buffer (with 20% [v/v], dimethylformamide) containing H 2 O 2 , with microfluidic transport enabled by electroosmotic flow (EOF). Michaelis–Menten kinetics were obtained with K m and V max values of 0.98 m M and 0.21 μmol H 2 O 2 converted/mg SBP per minute, respectively. These values are nearly identical to nonimmobilized SBP kinetics in aqueous–DMF solutions in 20‐μL volumes in 384‐well plates and 5‐mL reaction volumes in 20‐mL scintillation vials. These results indicate that SBP displays intrinsically native activity even in the immobilized form at the microscale, and further attests to the mild immobilization conditions afforded by PMA. Bienzymic and trienzymic reactions were also performed in the microfluidic biochip. Specifically, a combined Candida antarctica lipase B–SBP bienzymic system was used to convert tolyl acetate into poly( p ‐cresol), and an invertase–glucose oxidase SBP trienzymic system was used to take sucrose and generate H 2 O 2 for SBP‐catalyzed synthesis of poly( p ‐cresol). © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 83: 20–28, 2003. Article in Journal/Newspaper Antarc* Antarctica Wiley Online Library Biotechnology and Bioengineering 83 1 20 28 |
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English |
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Abstract The attachment of enzymes to glass microfluidic channels has been achieved using a highly reactive poly(maleic anhydride‐ alt ‐α‐olefin) (PMA)‐based coating that is supplied to the microchannel in a toluene solution. The PMA reacts with 3‐aminopropyltriethoxysilane groups linked to the glass surface to form a matrix that enables additional maleic anhydride groups to react with free amino groups on enzymes to give a mixed covalent–noncovalent immobilization support. Using a simple T‐channel microfluidic design, with reaction channel dimensions of 200 μm wide (at the center), 15 μm deep, and 30 mm long giving a reaction volume of 90 nL, soybean peroxidase (SBP) was attached at an amount up to 0.6 μg/channel. SBP‐catalyzed oxidation of p ‐cresol was performed in aqueous buffer (with 20% [v/v], dimethylformamide) containing H 2 O 2 , with microfluidic transport enabled by electroosmotic flow (EOF). Michaelis–Menten kinetics were obtained with K m and V max values of 0.98 m M and 0.21 μmol H 2 O 2 converted/mg SBP per minute, respectively. These values are nearly identical to nonimmobilized SBP kinetics in aqueous–DMF solutions in 20‐μL volumes in 384‐well plates and 5‐mL reaction volumes in 20‐mL scintillation vials. These results indicate that SBP displays intrinsically native activity even in the immobilized form at the microscale, and further attests to the mild immobilization conditions afforded by PMA. Bienzymic and trienzymic reactions were also performed in the microfluidic biochip. Specifically, a combined Candida antarctica lipase B–SBP bienzymic system was used to convert tolyl acetate into poly( p ‐cresol), and an invertase–glucose oxidase SBP trienzymic system was used to take sucrose and generate H 2 O 2 for SBP‐catalyzed synthesis of poly( p ‐cresol). © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 83: 20–28, 2003. |
format |
Article in Journal/Newspaper |
author |
Lee, Moo‐Yeal Srinivasan, Aravind Ku, Bosung Dordick, Jonathan S. |
spellingShingle |
Lee, Moo‐Yeal Srinivasan, Aravind Ku, Bosung Dordick, Jonathan S. Multienzyme catalysis in microfluidic biochips |
author_facet |
Lee, Moo‐Yeal Srinivasan, Aravind Ku, Bosung Dordick, Jonathan S. |
author_sort |
Lee, Moo‐Yeal |
title |
Multienzyme catalysis in microfluidic biochips |
title_short |
Multienzyme catalysis in microfluidic biochips |
title_full |
Multienzyme catalysis in microfluidic biochips |
title_fullStr |
Multienzyme catalysis in microfluidic biochips |
title_full_unstemmed |
Multienzyme catalysis in microfluidic biochips |
title_sort |
multienzyme catalysis in microfluidic biochips |
publisher |
Wiley |
publishDate |
2003 |
url |
http://dx.doi.org/10.1002/bit.10642 https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1002%2Fbit.10642 https://onlinelibrary.wiley.com/doi/pdf/10.1002/bit.10642 |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_source |
Biotechnology and Bioengineering volume 83, issue 1, page 20-28 ISSN 0006-3592 1097-0290 |
op_rights |
http://onlinelibrary.wiley.com/termsAndConditions#vor |
op_doi |
https://doi.org/10.1002/bit.10642 |
container_title |
Biotechnology and Bioengineering |
container_volume |
83 |
container_issue |
1 |
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20 |
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28 |
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1800741655774494720 |