Biochemical and structural characterization of cross-linked enzyme aggregates (CLEAs) of organic solvent tolerant protease
Cross-linked enzyme aggregates (CLEAs) is an immobilization technique that can be used to customize enzymes under an optimized condition. Structural analysis on any enzyme treated with a CLEA remains elusive and has been less explored. In the present work, a method for preparing an organic solvent t...
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| Format: | Article |
| Language: | English |
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MDPI
2020
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| Online Access: | http://psasir.upm.edu.my/id/eprint/38188/ http://psasir.upm.edu.my/id/eprint/38188/1/38188.pdf |
| _version_ | 1848848809913745408 |
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| author | Mohd Razib, Muhammad Syafiq Raja Abdul Rahman, Raja Noor Zaliha Mohd Shariff, Fairolniza Mohamad Ali, Mohd Shukuri |
| author_facet | Mohd Razib, Muhammad Syafiq Raja Abdul Rahman, Raja Noor Zaliha Mohd Shariff, Fairolniza Mohamad Ali, Mohd Shukuri |
| author_sort | Mohd Razib, Muhammad Syafiq |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | Cross-linked enzyme aggregates (CLEAs) is an immobilization technique that can be used to customize enzymes under an optimized condition. Structural analysis on any enzyme treated with a CLEA remains elusive and has been less explored. In the present work, a method for preparing an organic solvent tolerant protease using a CLEA is disclosed and optimized for better biochemical properties, followed by an analysis of the structure of this CLEA-treated protease. The said organic solvent tolerant protease is a metalloprotease known as elastase strain K in which activity of the metalloprotease is measured by a biochemical interaction with azocasein. Results showed that when a glutaraldehyde of 0.02% (v/v) was used under a 2 h treatment, the amount of recovered activity in CLEA-elastase was highest. The recovered activity of CLEA-elastase and CLEA-elastase-SB (which was a CLEA co-aggregated with starch and bovine serum albumin (BSA)) were at an approximate 60% and 80%, respectively. The CLEA immobilization of elastase strain K allowed the stability of the enzyme to be enhanced at high temperature and at a broader pH. Both CLEA-elastase and CLEA-elastase-SB end-products were able to maintain up to 67% enzyme activity at 60 °C and exhibiting an enhanced stability within pH 5–9 with up to 90% recovering activity. By implementing a CLEA on the organic solvent tolerant protease, the characteristics of the organic solvent tolerant were preserved and enhanced with the presence of 25% (v/v) acetonitrile, ethanol, and benzene at 165%, 173%, and 153% relative activity. Structural analysis through SEM and dynamic light scattering (DLS) showed that CLEA-elastase had a random aggregate morphology with an average diameter of 1497 nm. |
| first_indexed | 2025-11-15T09:40:24Z |
| format | Article |
| id | upm-38188 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T09:40:24Z |
| publishDate | 2020 |
| publisher | MDPI |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | upm-381882020-05-03T23:05:41Z http://psasir.upm.edu.my/id/eprint/38188/ Biochemical and structural characterization of cross-linked enzyme aggregates (CLEAs) of organic solvent tolerant protease Mohd Razib, Muhammad Syafiq Raja Abdul Rahman, Raja Noor Zaliha Mohd Shariff, Fairolniza Mohamad Ali, Mohd Shukuri Cross-linked enzyme aggregates (CLEAs) is an immobilization technique that can be used to customize enzymes under an optimized condition. Structural analysis on any enzyme treated with a CLEA remains elusive and has been less explored. In the present work, a method for preparing an organic solvent tolerant protease using a CLEA is disclosed and optimized for better biochemical properties, followed by an analysis of the structure of this CLEA-treated protease. The said organic solvent tolerant protease is a metalloprotease known as elastase strain K in which activity of the metalloprotease is measured by a biochemical interaction with azocasein. Results showed that when a glutaraldehyde of 0.02% (v/v) was used under a 2 h treatment, the amount of recovered activity in CLEA-elastase was highest. The recovered activity of CLEA-elastase and CLEA-elastase-SB (which was a CLEA co-aggregated with starch and bovine serum albumin (BSA)) were at an approximate 60% and 80%, respectively. The CLEA immobilization of elastase strain K allowed the stability of the enzyme to be enhanced at high temperature and at a broader pH. Both CLEA-elastase and CLEA-elastase-SB end-products were able to maintain up to 67% enzyme activity at 60 °C and exhibiting an enhanced stability within pH 5–9 with up to 90% recovering activity. By implementing a CLEA on the organic solvent tolerant protease, the characteristics of the organic solvent tolerant were preserved and enhanced with the presence of 25% (v/v) acetonitrile, ethanol, and benzene at 165%, 173%, and 153% relative activity. Structural analysis through SEM and dynamic light scattering (DLS) showed that CLEA-elastase had a random aggregate morphology with an average diameter of 1497 nm. MDPI 2020 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/38188/1/38188.pdf Mohd Razib, Muhammad Syafiq and Raja Abdul Rahman, Raja Noor Zaliha and Mohd Shariff, Fairolniza and Mohamad Ali, Mohd Shukuri (2020) Biochemical and structural characterization of cross-linked enzyme aggregates (CLEAs) of organic solvent tolerant protease. Catalysts, 10 (1). art. no. 55. pp. 1-16. ISSN 2073-4344 https://www.mdpi.com/2073-4344/10/1/55 10.3390/catal10010055 |
| spellingShingle | Mohd Razib, Muhammad Syafiq Raja Abdul Rahman, Raja Noor Zaliha Mohd Shariff, Fairolniza Mohamad Ali, Mohd Shukuri Biochemical and structural characterization of cross-linked enzyme aggregates (CLEAs) of organic solvent tolerant protease |
| title | Biochemical and structural characterization of cross-linked enzyme aggregates (CLEAs) of organic solvent tolerant protease |
| title_full | Biochemical and structural characterization of cross-linked enzyme aggregates (CLEAs) of organic solvent tolerant protease |
| title_fullStr | Biochemical and structural characterization of cross-linked enzyme aggregates (CLEAs) of organic solvent tolerant protease |
| title_full_unstemmed | Biochemical and structural characterization of cross-linked enzyme aggregates (CLEAs) of organic solvent tolerant protease |
| title_short | Biochemical and structural characterization of cross-linked enzyme aggregates (CLEAs) of organic solvent tolerant protease |
| title_sort | biochemical and structural characterization of cross-linked enzyme aggregates (cleas) of organic solvent tolerant protease |
| url | http://psasir.upm.edu.my/id/eprint/38188/ http://psasir.upm.edu.my/id/eprint/38188/ http://psasir.upm.edu.my/id/eprint/38188/ http://psasir.upm.edu.my/id/eprint/38188/1/38188.pdf |