Improvement of cross-linking and stability on cross-linked enzyme aggregate (CLEA)-xylanase by protein surface engineering
Autor: | Hesham A. El-Enshasy, Jamaliah Md Jahim, Abdul Munir Abdul Murad, Mohd Khairul Hakimi bin Abdul Wahab, Farah Diba Abu Bakar, Rosli Md. Illias |
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Rok vydání: | 2019 |
Předmět: |
0106 biological sciences
chemistry.chemical_classification 0303 health sciences Cross-linked enzyme aggregate Immobilized enzyme Bioengineering 01 natural sciences Applied Microbiology and Biotechnology Biochemistry Combinatorial chemistry Catalysis 03 medical and health sciences chemistry.chemical_compound Enzyme chemistry Biocatalysis 010608 biotechnology Xylanase Surface modification Glutaraldehyde 030304 developmental biology |
Zdroj: | Process Biochemistry. 86:40-49 |
ISSN: | 1359-5113 |
DOI: | 10.1016/j.procbio.2019.07.017 |
Popis: | 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. |
Databáze: | OpenAIRE |
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