Silica mesoporous materials were synthesized and used as the supporting materials for immobilization of enzyme.Penicillin G acylase,an enzyme which was used to produce 6-APA in pharmaceutical industry,was immobilized ...Silica mesoporous materials were synthesized and used as the supporting materials for immobilization of enzyme.Penicillin G acylase,an enzyme which was used to produce 6-APA in pharmaceutical industry,was immobilized in the mesoporous materials by the immersion method.The stabilities of the immobilized penicillin G acylase were studied.After incubation at 60 ℃ for 2 h,the activity of immobilized penicillin G acylase remained 80% in the best case.At higher or lower pH,the free enzyme was deactivated quickly,while the immobilized enzyme still retained active.The result of operational stability showed that the immobilized(enzyme) retained 70% of its initial activity after operating for 6 times.These results showed that the stabilities of immobilized penicillin G acylase,related to the pore size of mesoporous materials,were increased significantly compared with those of free enzyme.The improvement of stabilities of immobilized enzyme was significant when the pore size of the mesoporous materials matched the enzyme molecule size.展开更多
Penicillin G acylase(PGA) was immobilized on the magnetic hydrophilic polymer microspheres with average pore size of 17.1 nm,specific surface area of 128.2 m2/g and saturate magnetization of 6.4 emu/g.The 96.7%ampicil...Penicillin G acylase(PGA) was immobilized on the magnetic hydrophilic polymer microspheres with average pore size of 17.1 nm,specific surface area of 128.2 m2/g and saturate magnetization of 6.4 emu/g.The 96.7%ampicillin yield with 1.60 of the synthesis/hydrolysis(S/H) ratio from 6-aminopenicillanic acid(6-APA) and D-(-)-alpha-phenylglycine methyl ester(D-PGME) can be achieved using the resultant magnetic biocatalyst in ethylene glycol,where only 82.1%yield with 1.40 of the S/H ratio was obtained using the free PGA under the identical reaction conditions.The immobilized PGA can be separated magnetically and recycled for five times without obvious loss of its catalytic activity.展开更多
通过研究氨苄西林、6-氨基青霉烷酸(6-APA)和D-苯甘氨酸甲酯(D-PGME)在聚乙二醇(PEG)和不同盐组成的双水相体系(ATPS)中的分配行为,建立了一个由PEG-4000(w=20%)和硫酸铵(w=15%)组成的双水相体系.在此体系中,氨苄西林的分配系数为5.0,6-...通过研究氨苄西林、6-氨基青霉烷酸(6-APA)和D-苯甘氨酸甲酯(D-PGME)在聚乙二醇(PEG)和不同盐组成的双水相体系(ATPS)中的分配行为,建立了一个由PEG-4000(w=20%)和硫酸铵(w=15%)组成的双水相体系.在此体系中,氨苄西林的分配系数为5.0,6-APA和D-PGME的分配系数分别为1.7和1.4.以环氧基功能化介孔分子筛SBA-15为载体固定青霉素酰化酶(Penicillin G Acylase,PGA),并在双水相体系中催化合成氨苄西林,产率为80%,经过4批次的连续合成,产率提高到98%,而在水相体系中酶促合成氨苄西林的产率仅为27%.展开更多
A hydrotalcite-like Mg 2+ /Al 3+ layered double hydroxide(LDH) material was prepared by means of a modified coprecipitation method involving a rapid mixing step followed by a separate aging process. LDH calci...A hydrotalcite-like Mg 2+ /Al 3+ layered double hydroxide(LDH) material was prepared by means of a modified coprecipitation method involving a rapid mixing step followed by a separate aging process. LDH calcined at 500 ℃, denoted as CLDH, was characterized by XRD, IR and BET surface area measurements. CLDH has a poor crystalline MgO-like structure with a high surface area and porosity. CLDH was used as a support for the immobilization of penicillin G acylase(PGA). The effect of varying the immobilization conditions, such as pH, contact time and the ratio of enzyme to support, on the activity of the immobilized enzyme in the hydrolysis of penicillin G has been studied. It was found that the activity of the immobilized enzyme decreased slightly with decreasing pH and reached a maximum after a contact time of 24 h. The activity of the immobilized enzyme increased with increasing the ratio of enzyme to support. It was found that the adsorption of PGA inhibited the expected reaction of CLDH with an aqueous medium to regenerate a LDH phase. Its original activity(36%) after 15 cycles of reuse of the immobilized enzyme was retained, but no further loss in the activity was observed.展开更多
文摘Silica mesoporous materials were synthesized and used as the supporting materials for immobilization of enzyme.Penicillin G acylase,an enzyme which was used to produce 6-APA in pharmaceutical industry,was immobilized in the mesoporous materials by the immersion method.The stabilities of the immobilized penicillin G acylase were studied.After incubation at 60 ℃ for 2 h,the activity of immobilized penicillin G acylase remained 80% in the best case.At higher or lower pH,the free enzyme was deactivated quickly,while the immobilized enzyme still retained active.The result of operational stability showed that the immobilized(enzyme) retained 70% of its initial activity after operating for 6 times.These results showed that the stabilities of immobilized penicillin G acylase,related to the pore size of mesoporous materials,were increased significantly compared with those of free enzyme.The improvement of stabilities of immobilized enzyme was significant when the pore size of the mesoporous materials matched the enzyme molecule size.
基金supported by The National Basic Research Program 973 of China(No.2010CB534916)the National Natural Science Foundation of China(Nos.50662004,20564002).
文摘Penicillin G acylase(PGA) was immobilized on the magnetic hydrophilic polymer microspheres with average pore size of 17.1 nm,specific surface area of 128.2 m2/g and saturate magnetization of 6.4 emu/g.The 96.7%ampicillin yield with 1.60 of the synthesis/hydrolysis(S/H) ratio from 6-aminopenicillanic acid(6-APA) and D-(-)-alpha-phenylglycine methyl ester(D-PGME) can be achieved using the resultant magnetic biocatalyst in ethylene glycol,where only 82.1%yield with 1.40 of the S/H ratio was obtained using the free PGA under the identical reaction conditions.The immobilized PGA can be separated magnetically and recycled for five times without obvious loss of its catalytic activity.
文摘通过研究氨苄西林、6-氨基青霉烷酸(6-APA)和D-苯甘氨酸甲酯(D-PGME)在聚乙二醇(PEG)和不同盐组成的双水相体系(ATPS)中的分配行为,建立了一个由PEG-4000(w=20%)和硫酸铵(w=15%)组成的双水相体系.在此体系中,氨苄西林的分配系数为5.0,6-APA和D-PGME的分配系数分别为1.7和1.4.以环氧基功能化介孔分子筛SBA-15为载体固定青霉素酰化酶(Penicillin G Acylase,PGA),并在双水相体系中催化合成氨苄西林,产率为80%,经过4批次的连续合成,产率提高到98%,而在水相体系中酶促合成氨苄西林的产率仅为27%.
基金Supported by the National Natural Science Foundation of China( No.2 99730 0 4 )
文摘A hydrotalcite-like Mg 2+ /Al 3+ layered double hydroxide(LDH) material was prepared by means of a modified coprecipitation method involving a rapid mixing step followed by a separate aging process. LDH calcined at 500 ℃, denoted as CLDH, was characterized by XRD, IR and BET surface area measurements. CLDH has a poor crystalline MgO-like structure with a high surface area and porosity. CLDH was used as a support for the immobilization of penicillin G acylase(PGA). The effect of varying the immobilization conditions, such as pH, contact time and the ratio of enzyme to support, on the activity of the immobilized enzyme in the hydrolysis of penicillin G has been studied. It was found that the activity of the immobilized enzyme decreased slightly with decreasing pH and reached a maximum after a contact time of 24 h. The activity of the immobilized enzyme increased with increasing the ratio of enzyme to support. It was found that the adsorption of PGA inhibited the expected reaction of CLDH with an aqueous medium to regenerate a LDH phase. Its original activity(36%) after 15 cycles of reuse of the immobilized enzyme was retained, but no further loss in the activity was observed.
