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Production of Galactooligosaccharides Using β-Galactosidase Immobilized on Chitosan-Coated Magnetic Nanoparticles with Tris(hydroxymethyl)phosphine as an Optional Coupling Agent.

Chen SC, Duan KJ - Int J Mol Sci (2015)

Bottom Line: However, activity retention of batchwise reactions was similar for both immobilized systems.All the three enzyme systems produced GOS compound with similar concentration profiles, with a maximum GOS yield of 50.5% from 36% (w · v(-1)) lactose on a dry weight basis.The chitosan-coated magnetic Fe3O4 nanoparticles can be regenerated using a desorption/re-adsorption process described in this study.

View Article: PubMed Central - PubMed

Affiliation: Department of Bioengineering, Tatung University, Taipei 104, Taiwan. monica94707@yahoo.com.tw.

ABSTRACT
β-Galactosidase was immobilized on chitosan-coated magnetic Fe3O4 nanoparticles and was used to produce galactooligosaccharides (GOS) from lactose. Immobilized enzyme was prepared with or without the coupling agent, tris(hydroxymethyl)phosphine (THP). The two immobilized systems and the free enzyme achieved their maximum activity at pH 6.0 with an optimal temperature of 50 °C. The immobilized enzymes showed higher activities at a wider range of temperatures and pH. Furthermore, the immobilized enzyme coupled with THP showed higher thermal stability than that without THP. However, activity retention of batchwise reactions was similar for both immobilized systems. All the three enzyme systems produced GOS compound with similar concentration profiles, with a maximum GOS yield of 50.5% from 36% (w · v(-1)) lactose on a dry weight basis. The chitosan-coated magnetic Fe3O4 nanoparticles can be regenerated using a desorption/re-adsorption process described in this study.

No MeSH data available.


Effect of pH on the relative enzyme activity at 50 °C. Fe3O4-CS-immobilized enzyme (0.2 g), Fe3O4-CS-THP-immobilized enzyme (0.2 g) or the free enzyme (1 mL) was added to 100 mL of 5% (w·v−1) lactose at 50 °C in a 500 mL Erlenmeyer flask and incubated at various pH from 3.0 to 8.0 in an orbital shaker bath at 200 rpm for 10 min. The relative activity was determined by measuring the production of glucose. The buffer used in this set of reactions were 0.1 M sodium acetate buffer (pH 3.0~5.6) and 0.1 M potassium dihydrogen orthophosphate (pH 5.8~8.0). The data are averaged from three samples. (●) Fe3O4-CS-immobilized enzyme; (○) Fe3O4-CS-THP-immobilized enzyme; (▼) free enzyme.
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ijms-16-12499-f004: Effect of pH on the relative enzyme activity at 50 °C. Fe3O4-CS-immobilized enzyme (0.2 g), Fe3O4-CS-THP-immobilized enzyme (0.2 g) or the free enzyme (1 mL) was added to 100 mL of 5% (w·v−1) lactose at 50 °C in a 500 mL Erlenmeyer flask and incubated at various pH from 3.0 to 8.0 in an orbital shaker bath at 200 rpm for 10 min. The relative activity was determined by measuring the production of glucose. The buffer used in this set of reactions were 0.1 M sodium acetate buffer (pH 3.0~5.6) and 0.1 M potassium dihydrogen orthophosphate (pH 5.8~8.0). The data are averaged from three samples. (●) Fe3O4-CS-immobilized enzyme; (○) Fe3O4-CS-THP-immobilized enzyme; (▼) free enzyme.

Mentions: The results of relative activity from the immobilized and free enzymes corresponding to pH changes are shown in Figure 4. The activities of Fe3O4-CS-THP, Fe3O4-CS-immobilized enzyme and the free enzymes were maximally achieved at pH 6.0. This result suggests that there was no apparent charge in the immobilization at the active site of the enzyme after coupling with THP. Both the immobilized enzymes show a little improvement of relative pH activity at pH 3.0 to 8.0.


Production of Galactooligosaccharides Using β-Galactosidase Immobilized on Chitosan-Coated Magnetic Nanoparticles with Tris(hydroxymethyl)phosphine as an Optional Coupling Agent.

Chen SC, Duan KJ - Int J Mol Sci (2015)

Effect of pH on the relative enzyme activity at 50 °C. Fe3O4-CS-immobilized enzyme (0.2 g), Fe3O4-CS-THP-immobilized enzyme (0.2 g) or the free enzyme (1 mL) was added to 100 mL of 5% (w·v−1) lactose at 50 °C in a 500 mL Erlenmeyer flask and incubated at various pH from 3.0 to 8.0 in an orbital shaker bath at 200 rpm for 10 min. The relative activity was determined by measuring the production of glucose. The buffer used in this set of reactions were 0.1 M sodium acetate buffer (pH 3.0~5.6) and 0.1 M potassium dihydrogen orthophosphate (pH 5.8~8.0). The data are averaged from three samples. (●) Fe3O4-CS-immobilized enzyme; (○) Fe3O4-CS-THP-immobilized enzyme; (▼) free enzyme.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4490457&req=5

ijms-16-12499-f004: Effect of pH on the relative enzyme activity at 50 °C. Fe3O4-CS-immobilized enzyme (0.2 g), Fe3O4-CS-THP-immobilized enzyme (0.2 g) or the free enzyme (1 mL) was added to 100 mL of 5% (w·v−1) lactose at 50 °C in a 500 mL Erlenmeyer flask and incubated at various pH from 3.0 to 8.0 in an orbital shaker bath at 200 rpm for 10 min. The relative activity was determined by measuring the production of glucose. The buffer used in this set of reactions were 0.1 M sodium acetate buffer (pH 3.0~5.6) and 0.1 M potassium dihydrogen orthophosphate (pH 5.8~8.0). The data are averaged from three samples. (●) Fe3O4-CS-immobilized enzyme; (○) Fe3O4-CS-THP-immobilized enzyme; (▼) free enzyme.
Mentions: The results of relative activity from the immobilized and free enzymes corresponding to pH changes are shown in Figure 4. The activities of Fe3O4-CS-THP, Fe3O4-CS-immobilized enzyme and the free enzymes were maximally achieved at pH 6.0. This result suggests that there was no apparent charge in the immobilization at the active site of the enzyme after coupling with THP. Both the immobilized enzymes show a little improvement of relative pH activity at pH 3.0 to 8.0.

Bottom Line: However, activity retention of batchwise reactions was similar for both immobilized systems.All the three enzyme systems produced GOS compound with similar concentration profiles, with a maximum GOS yield of 50.5% from 36% (w · v(-1)) lactose on a dry weight basis.The chitosan-coated magnetic Fe3O4 nanoparticles can be regenerated using a desorption/re-adsorption process described in this study.

View Article: PubMed Central - PubMed

Affiliation: Department of Bioengineering, Tatung University, Taipei 104, Taiwan. monica94707@yahoo.com.tw.

ABSTRACT
β-Galactosidase was immobilized on chitosan-coated magnetic Fe3O4 nanoparticles and was used to produce galactooligosaccharides (GOS) from lactose. Immobilized enzyme was prepared with or without the coupling agent, tris(hydroxymethyl)phosphine (THP). The two immobilized systems and the free enzyme achieved their maximum activity at pH 6.0 with an optimal temperature of 50 °C. The immobilized enzymes showed higher activities at a wider range of temperatures and pH. Furthermore, the immobilized enzyme coupled with THP showed higher thermal stability than that without THP. However, activity retention of batchwise reactions was similar for both immobilized systems. All the three enzyme systems produced GOS compound with similar concentration profiles, with a maximum GOS yield of 50.5% from 36% (w · v(-1)) lactose on a dry weight basis. The chitosan-coated magnetic Fe3O4 nanoparticles can be regenerated using a desorption/re-adsorption process described in this study.

No MeSH data available.