Improvement of cross-linking and stability on cross-linked enzyme aggregate (CLEA)-xylanase by protein surface engineering
Cross-linked enzyme aggregate (CLEA) is a well-known enzyme immobilization technique that is efficient and cost-effective. In this study, we proposed a combination of the surface modification of an enzyme and CLEA to increase the number of free amino groups. Site-directed mutagenesis was applied to...
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Elsevier Ltd
2019
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| author | Abdul Wahab, Mohd. Khairul Hakimi El-Enshasy, Hesham Ali Abu Bakar, Farah Diba Abdul Murad, Abdul Munir Md. Jahim, Jamaliah Md. Illias, Rosli |
| author_facet | Abdul Wahab, Mohd. Khairul Hakimi El-Enshasy, Hesham Ali Abu Bakar, Farah Diba Abdul Murad, Abdul Munir Md. Jahim, Jamaliah Md. Illias, Rosli |
| author_sort | Abdul Wahab, Mohd. Khairul Hakimi |
| collection | ePrints |
| description | Cross-linked enzyme aggregate (CLEA) is a well-known enzyme immobilization technique that is efficient and cost-effective. In this study, we proposed a combination of the surface modification of an enzyme and CLEA to increase the number of free amino groups. Site-directed mutagenesis was applied to selected residues of xylanase from Aspergillus fumigatus RT-1 and the xylanase was subsequently cross-linked using glutaraldehyde. Surface analysis of the xylanase revealed that 9 residues were exposed to the environment and only 3 were lysines. Thus, four additional lysines were substituted for residues opposite of the catalytic region. After optimizing the CLEA parameters, a stable cross-linked mutant xylanase with the addition of BSA (mXyn-CLEA-BSA) was obtained where the enzyme was 1.09-fold, 1.35-fold and 1.77-fold more stable than the cross-linked recombinant xylanase with the addition of BSA (rXyn-CLEA-BSA), without the addition of BSA (rXyn-CLEA) and free enzyme (rXyn), respectively. In terms of reusability, rXyn-CLEA can be used up to 5 cycles, rXyn-CLEA-BSA and mXyn-CLEA up to 7 cycles and mXyn-CLEA-BSA up to 8 cycles until the total activity is lost. The increase in stability and reusability using this approach provides a promising biocatalyst that can be further utilized in the production of prebiotics in the biomass industry. |
| first_indexed | 2024-03-05T20:48:06Z |
| format | Article |
| id | utm.eprints-89547 |
| institution | Universiti Teknologi Malaysia - ePrints |
| last_indexed | 2024-03-05T20:48:06Z |
| publishDate | 2019 |
| publisher | Elsevier Ltd |
| record_format | dspace |
| spelling | utm.eprints-895472021-02-22T06:08:14Z http://eprints.utm.my/89547/ Improvement of cross-linking and stability on cross-linked enzyme aggregate (CLEA)-xylanase by protein surface engineering Abdul Wahab, Mohd. Khairul Hakimi El-Enshasy, Hesham Ali Abu Bakar, Farah Diba Abdul Murad, Abdul Munir Md. Jahim, Jamaliah Md. Illias, Rosli TP Chemical technology Cross-linked enzyme aggregate (CLEA) is a well-known enzyme immobilization technique that is efficient and cost-effective. In this study, we proposed a combination of the surface modification of an enzyme and CLEA to increase the number of free amino groups. Site-directed mutagenesis was applied to selected residues of xylanase from Aspergillus fumigatus RT-1 and the xylanase was subsequently cross-linked using glutaraldehyde. Surface analysis of the xylanase revealed that 9 residues were exposed to the environment and only 3 were lysines. Thus, four additional lysines were substituted for residues opposite of the catalytic region. After optimizing the CLEA parameters, a stable cross-linked mutant xylanase with the addition of BSA (mXyn-CLEA-BSA) was obtained where the enzyme was 1.09-fold, 1.35-fold and 1.77-fold more stable than the cross-linked recombinant xylanase with the addition of BSA (rXyn-CLEA-BSA), without the addition of BSA (rXyn-CLEA) and free enzyme (rXyn), respectively. In terms of reusability, rXyn-CLEA can be used up to 5 cycles, rXyn-CLEA-BSA and mXyn-CLEA up to 7 cycles and mXyn-CLEA-BSA up to 8 cycles until the total activity is lost. The increase in stability and reusability using this approach provides a promising biocatalyst that can be further utilized in the production of prebiotics in the biomass industry. Elsevier Ltd 2019-11 Article PeerReviewed Abdul Wahab, Mohd. Khairul Hakimi and El-Enshasy, Hesham Ali and Abu Bakar, Farah Diba and Abdul Murad, Abdul Munir and Md. Jahim, Jamaliah and Md. Illias, Rosli (2019) Improvement of cross-linking and stability on cross-linked enzyme aggregate (CLEA)-xylanase by protein surface engineering. Process Biochemistry, 86 . pp. 40-49. ISSN 1359-5113 http://dx.doi.org/10.1016/j.procbio.2019.07.017 DOI:10.1016/j.procbio.2019.07.017 |
| spellingShingle | TP Chemical technology Abdul Wahab, Mohd. Khairul Hakimi El-Enshasy, Hesham Ali Abu Bakar, Farah Diba Abdul Murad, Abdul Munir Md. Jahim, Jamaliah Md. Illias, Rosli Improvement of cross-linking and stability on cross-linked enzyme aggregate (CLEA)-xylanase by protein surface engineering |
| title | Improvement of cross-linking and stability on cross-linked enzyme aggregate (CLEA)-xylanase by protein surface engineering |
| title_full | Improvement of cross-linking and stability on cross-linked enzyme aggregate (CLEA)-xylanase by protein surface engineering |
| title_fullStr | Improvement of cross-linking and stability on cross-linked enzyme aggregate (CLEA)-xylanase by protein surface engineering |
| title_full_unstemmed | Improvement of cross-linking and stability on cross-linked enzyme aggregate (CLEA)-xylanase by protein surface engineering |
| title_short | Improvement of cross-linking and stability on cross-linked enzyme aggregate (CLEA)-xylanase by protein surface engineering |
| title_sort | improvement of cross linking and stability on cross linked enzyme aggregate clea xylanase by protein surface engineering |
| topic | TP Chemical technology |
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