Polyethylenimine-Assisted Aggregation of β-Galactosidase from Kluyveromyces lactis

Nedim Albayrak

Article Details

Polyethylenimine-Assisted Aggregation of β-Galactosidase from Kluyveromyces lactis

Authors:

Nedim Albayrak

Publisher:

Hitit University

Main Topics:

Fermentation Technology | Food Biotechnology

Journal:

Hittite Journal of Science and Engineering Vol.11 Issue 4, pp.149-156

Document Type:

Article

Publication Date:

12/2024

Document Language:

English

Document ID (Org No.):

2001337652

pISSN:

2148-4171

Keywords:

Abstract:

 Using PEI as an aggregating agent with β-galactosidase from Kluyveromyces lactis was investigated with no solid support. PEI, a cationic polymer initiated instant aggregation with the enzyme in solution. The factors affecting the aggregation such as PEI to enzyme ratio, glutar aldehyde for cross-linking and pH were investigated. Aggregation and sedimentation as well as the residual activity of aggregates were effectively dependent on the PEI to enzyme ratios. Easily precipitating aggregates at the ratio of 18, PEI enzyme aggregates were able to contain all enzyme in the complexation and displayed 60 of initial lactase activity. The PEI aggregation of enzyme led to enhancements in chemical and physical characteristics compered to free enzyme. The soluble enzyme showed a narrow optimum at about pH 7 while pH optimum of the aggregates extended one pH unit toward the alkaline range. Upon overnight incubation at 40 ºC, aggregated enzyme displayed 30 greater stability on average at all pHs tested. Although the free enzyme showed the highest activity at 40C, it rapidly lost 50 of its activity at 50C. In contrast, the aggregated enzyme retained full activity at 50C and 70 activity at 65C. With markedly enhanced thermal stability, the half-life of the aggregated enzyme increased from 76 hours to 254 hours at 40ºC. Overall, the simple and rapid aggregation of PEI with the enzyme led to instant and intense clustering, resulting in higher thermal and pH stabilities. This method may potentially offer efficient and cost-effective catalysis in lactase conversion processes.

 Using PEI as an aggregating agent with β-galactosidase from Kluyveromyces lactis was investigated with no solid support. PEI, a cationic polymer initiated instant aggregation with the enzyme in solution. The factors affecting the aggregation such as PEI to enzyme ratio, glutar aldehyde for cross-linking and pH were investigated. Aggregation and sedimentation as well as the residual activity of aggregates were effectively dependent on the PEI to enzyme ratios. Easily precipitating aggregates at the ratio of 18, PEI enzyme aggregates were able to contain all enzyme in the complexation and displayed 60 of initial lactase activity. The PEI aggregation of enzyme led to enhancements in chemical and physical characteristics compered to free enzyme. The soluble enzyme showed a narrow optimum at about pH 7 while pH optimum of the aggregates extended one pH unit toward the alkaline range. Upon overnight incubation at 40 ºC, aggregated enzyme displayed 30 greater stability on average at all pHs tested. Although the free enzyme showed the highest activity at 40C, it rapidly lost 50 of its activity at 50C. In contrast, the aggregated enzyme retained full activity at 50C and 70 activity at 65C. With markedly enhanced thermal stability, the half-life of the aggregated enzyme increased from 76 hours to 254 hours at 40ºC. Overall, the simple and rapid aggregation of PEI with the enzyme led to instant and intense clustering, resulting in higher thermal and pH stabilities. This method may potentially offer efficient and cost-effective catalysis in lactase conversion processes.

 Using PEI as an aggregating agent with β-galactosidase from Kluyveromyces lactis was investigated with no solid support. PEI, a cationic polymer initiated instant aggregation with the enzyme in solution. The factors affecting the aggregation such as PEI to enzyme ratio, glutar aldehyde for cross-linking and pH were investigated. Aggregation and sedimentation as well as the residual activity of aggregates were effectively dependent on the PEI to enzyme ratios. Easily precipitating aggregates at the ratio of 18, PEI enzyme aggregates were able to contain all enzyme in the complexation and displayed 60 of initial lactase activity. The PEI aggregation of enzyme led to enhancements in chemical and physical characteristics compered to free enzyme. The soluble enzyme showed a narrow optimum at about pH 7 while pH optimum of the aggregates extended one pH unit toward the alkaline range. Upon overnight incubation at 40 ºC, aggregated enzyme displayed 30 greater stability on average at all pHs tested. Although the free enzyme showed the highest activity at 40C, it rapidly lost 50 of its activity at 50C. In contrast, the aggregated enzyme retained full activity at 50C and 70 activity at 65C. With markedly enhanced thermal stability, the half-life of the aggregated enzyme increased from 76 hours to 254 hours at 40ºC. Overall, the simple and rapid aggregation of PEI with the enzyme led to instant and intense clustering, resulting in higher thermal and pH stabilities. This method may potentially offer efficient and cost-effective catalysis in lactase conversion processes.

 Using PEI as an aggregating agent with β-galactosidase from Kluyveromyces lactis was investigated with no solid support. PEI, a cationic polymer initiated instant aggregation with the enzyme in solution. The factors affecting the aggregation such as PEI to enzyme ratio, glutar aldehyde for cross-linking and pH were investigated. Aggregation and sedimentation as well as the residual activity of aggregates were effectively dependent on the PEI to enzyme ratios. Easily precipitating aggregates at the ratio of 18, PEI enzyme aggregates were able to contain all enzyme in the complexation and displayed 60 of initial lactase activity. The PEI aggregation of enzyme led to enhancements in chemical and physical characteristics compered to free enzyme. The soluble enzyme showed a narrow optimum at about pH 7 while pH optimum of the aggregates extended one pH unit toward the alkaline range. Upon overnight incubation at 40 ºC, aggregated enzyme displayed 30 greater stability on average at all pHs tested. Although the free enzyme showed the highest activity at 40C, it rapidly lost 50 of its activity at 50C. In contrast, the aggregated enzyme retained full activity at 50C and 70 activity at 65C. With markedly enhanced thermal stability, the half-life of the aggregated enzyme increased from 76 hours to 254 hours at 40ºC. Overall, the simple and rapid aggregation of PEI with the enzyme led to instant and intense clustering, resulting in higher thermal and pH stabilities. This method may potentially offer efficient and cost-effective catalysis in lactase conversion processes.

Article Details

Polyethylenimine-Assisted Aggregation of β-Galactosidase from Kluyveromyces lactis

Authors:

Nedim Albayrak

Publisher:

Hitit University

Main Topics:

Fermentation Technology | Food Biotechnology

Journal:

Hittite Journal of Science and Engineering Vol.11 Issue 4, pp.149-156

Document Type:

Article

Publication Date:

12/2024

Document Language:

English

Document ID (Org No.):

2001337652

pISSN:

2148-4171

Kluyveromyces lactis

Lactase

Carrier-Free

PEI-Enzyme Aggregation

Stabilization

PEI

GA

Issues

Article Details

Authors: Nedim Albayrak

Publisher: Hitit University

Main Topics: Fermentation Technology | Food Biotechnology

Journal: Hittite Journal of Science and Engineering Vol.11 Issue 4, pp.149-156

Document Type: Article

Publication Date: 12/2024

Document Language: English

Document ID (Org No.): 2001337652

pISSN: 2148-4171

Issues

Authors:

Nedim Albayrak

Publisher:

Hitit University

Main Topics:

Fermentation Technology | Food Biotechnology

Journal:

Hittite Journal of Science and Engineering Vol.11 Issue 4, pp.149-156

Document Type:

Article

Publication Date:

12/2024

Document Language:

English

Document ID (Org No.):

2001337652

pISSN:

2148-4171