Production and Characterization of Cross-Linked Aggregates of <i>Geobacillus thermoleovorans</i> CCR11 Thermoalkaliphilic Recombinant Lipase
Immobilization of enzymes has many advantages for their application in biotechnological processes. In particular, the cross-linked enzyme aggregates (CLEAs) allow the production of solid biocatalysts with a high enzymatic loading and the advantage of obtaining derivatives with high stability at low...
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| Format: | Article |
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MDPI AG
2021-12-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/26/24/7569 |
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| author | Rosa-María Oliart-Ros Giselle-Lilian Badillo-Zeferino Rodolfo Quintana-Castro Irving-Israel Ruíz-López Alfonso Alexander-Aguilera Jorge-Guillermo Domínguez-Chávez Azmat Ali Khan Dinh Duc Nguyen Ashok Kumar Nadda María-Guadalupe Sánchez-Otero |
| author_facet | Rosa-María Oliart-Ros Giselle-Lilian Badillo-Zeferino Rodolfo Quintana-Castro Irving-Israel Ruíz-López Alfonso Alexander-Aguilera Jorge-Guillermo Domínguez-Chávez Azmat Ali Khan Dinh Duc Nguyen Ashok Kumar Nadda María-Guadalupe Sánchez-Otero |
| author_sort | Rosa-María Oliart-Ros |
| collection | DOAJ |
| description | Immobilization of enzymes has many advantages for their application in biotechnological processes. In particular, the cross-linked enzyme aggregates (CLEAs) allow the production of solid biocatalysts with a high enzymatic loading and the advantage of obtaining derivatives with high stability at low cost. The purpose of this study was to produce cross-linked enzymatic aggregates (CLEAs) of LipMatCCR11, a 43 kDa recombinant solvent-tolerant thermoalkaliphilic lipase from <i>Geobacillus thermoleovorans</i> CCR11. LipMatCCR11-CLEAs were prepared using (NH<sub>4</sub>)<sub>2</sub>SO<sub>4</sub> (40% <i>w</i>/<i>v</i>) as precipitant agent and glutaraldehyde (40 mM) as cross-linker, at pH 9, 20 °C. A U<sub>10</sub>(5<sup>6</sup>) uniform design was used to optimize CLEA production, varying protein concentration, ammonium sulfate %, pH, glutaraldehyde concentration, temperature, and incubation time. The synthesized CLEAs were also analyzed using scanning electron microscopy (SEM) that showed individual particles of <1 µm grouped to form a superstructure. The cross-linked aggregates showed a maximum mass activity of 7750 U/g at 40 °C and pH 8 and retained more than 20% activity at 100 °C. Greater thermostability, resistance to alkaline conditions and the presence of organic solvents, and better durability during storage were observed for LipMatCCR11-CLEAs in comparison with the soluble enzyme. LipMatCCR11-CLEAs presented good reusability by conserving 40% of their initial activity after 9 cycles of reuse. |
| first_indexed | 2024-03-10T03:28:10Z |
| format | Article |
| id | doaj.art-4b06ed08a00d4d399681a08c5872d359 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-10T03:28:10Z |
| publishDate | 2021-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-4b06ed08a00d4d399681a08c5872d3592023-11-23T09:46:06ZengMDPI AGMolecules1420-30492021-12-012624756910.3390/molecules26247569Production and Characterization of Cross-Linked Aggregates of <i>Geobacillus thermoleovorans</i> CCR11 Thermoalkaliphilic Recombinant LipaseRosa-María Oliart-Ros0Giselle-Lilian Badillo-Zeferino1Rodolfo Quintana-Castro2Irving-Israel Ruíz-López3Alfonso Alexander-Aguilera4Jorge-Guillermo Domínguez-Chávez5Azmat Ali Khan6Dinh Duc Nguyen7Ashok Kumar Nadda8María-Guadalupe Sánchez-Otero9Unidad de Investigación y Desarrollo en Alimentos, Tecnológico Nacional de México, Instituto Tecnológico de Veracruz, M.A. De Quevedo 2779, Veracruz C.P. 91897, Ver., MexicoUnidad de Investigación y Desarrollo en Alimentos, Tecnológico Nacional de México, Instituto Tecnológico de Veracruz, M.A. De Quevedo 2779, Veracruz C.P. 91897, Ver., MexicoFacultad de Bioanálisis, Universidad Veracruzana, Carmen Serdán Esq. Iturbide, Veracruz C.P. 91700, Ver., MexicoFacultad de Ingeniería Química, Benemérita Universidad Autónoma de Puebla, Av. San Claudio y 18 Sur, Ciudad Universitaria, Puebla C.P. 72570, Pue., MexicoFacultad de Bioanálisis, Universidad Veracruzana, Carmen Serdán Esq. Iturbide, Veracruz C.P. 91700, Ver., MexicoFacultad de Bioanálisis, Universidad Veracruzana, Carmen Serdán Esq. Iturbide, Veracruz C.P. 91700, Ver., MexicoPharmaceutical Biotechnology Laboratory, Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, Riyadh 11451, Saudi ArabiaDepartment of Environmental and Energy Engineering, Kyonggi University, 154-42 Gwanggyosan-ro, Yeongtong-gu, Suwon-si 16227, Gyeonggi-do, KoreaDepartment of Biotechnology and Bioinformatics, Faculty of Biotechnology, Jaypee University of Information Technology, Waknaghat, Solan, Himachal Pradesh 173 234, IndiaFacultad de Bioanálisis, Universidad Veracruzana, Carmen Serdán Esq. Iturbide, Veracruz C.P. 91700, Ver., MexicoImmobilization of enzymes has many advantages for their application in biotechnological processes. In particular, the cross-linked enzyme aggregates (CLEAs) allow the production of solid biocatalysts with a high enzymatic loading and the advantage of obtaining derivatives with high stability at low cost. The purpose of this study was to produce cross-linked enzymatic aggregates (CLEAs) of LipMatCCR11, a 43 kDa recombinant solvent-tolerant thermoalkaliphilic lipase from <i>Geobacillus thermoleovorans</i> CCR11. LipMatCCR11-CLEAs were prepared using (NH<sub>4</sub>)<sub>2</sub>SO<sub>4</sub> (40% <i>w</i>/<i>v</i>) as precipitant agent and glutaraldehyde (40 mM) as cross-linker, at pH 9, 20 °C. A U<sub>10</sub>(5<sup>6</sup>) uniform design was used to optimize CLEA production, varying protein concentration, ammonium sulfate %, pH, glutaraldehyde concentration, temperature, and incubation time. The synthesized CLEAs were also analyzed using scanning electron microscopy (SEM) that showed individual particles of <1 µm grouped to form a superstructure. The cross-linked aggregates showed a maximum mass activity of 7750 U/g at 40 °C and pH 8 and retained more than 20% activity at 100 °C. Greater thermostability, resistance to alkaline conditions and the presence of organic solvents, and better durability during storage were observed for LipMatCCR11-CLEAs in comparison with the soluble enzyme. LipMatCCR11-CLEAs presented good reusability by conserving 40% of their initial activity after 9 cycles of reuse.https://www.mdpi.com/1420-3049/26/24/7569cross-linked enzymatic aggregates (CLEAs)thermoalkaliphilic lipase<i>Geobacillus thermoleovorans</i> |
| spellingShingle | Rosa-María Oliart-Ros Giselle-Lilian Badillo-Zeferino Rodolfo Quintana-Castro Irving-Israel Ruíz-López Alfonso Alexander-Aguilera Jorge-Guillermo Domínguez-Chávez Azmat Ali Khan Dinh Duc Nguyen Ashok Kumar Nadda María-Guadalupe Sánchez-Otero Production and Characterization of Cross-Linked Aggregates of <i>Geobacillus thermoleovorans</i> CCR11 Thermoalkaliphilic Recombinant Lipase Molecules cross-linked enzymatic aggregates (CLEAs) thermoalkaliphilic lipase <i>Geobacillus thermoleovorans</i> |
| title | Production and Characterization of Cross-Linked Aggregates of <i>Geobacillus thermoleovorans</i> CCR11 Thermoalkaliphilic Recombinant Lipase |
| title_full | Production and Characterization of Cross-Linked Aggregates of <i>Geobacillus thermoleovorans</i> CCR11 Thermoalkaliphilic Recombinant Lipase |
| title_fullStr | Production and Characterization of Cross-Linked Aggregates of <i>Geobacillus thermoleovorans</i> CCR11 Thermoalkaliphilic Recombinant Lipase |
| title_full_unstemmed | Production and Characterization of Cross-Linked Aggregates of <i>Geobacillus thermoleovorans</i> CCR11 Thermoalkaliphilic Recombinant Lipase |
| title_short | Production and Characterization of Cross-Linked Aggregates of <i>Geobacillus thermoleovorans</i> CCR11 Thermoalkaliphilic Recombinant Lipase |
| title_sort | production and characterization of cross linked aggregates of i geobacillus thermoleovorans i ccr11 thermoalkaliphilic recombinant lipase |
| topic | cross-linked enzymatic aggregates (CLEAs) thermoalkaliphilic lipase <i>Geobacillus thermoleovorans</i> |
| url | https://www.mdpi.com/1420-3049/26/24/7569 |
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