Hybrid magnetic cross-linked enzyme aggregates of phenylalanine ammonia lyase from Rhodotorula glutinis.
Novel hybrid magnetic cross-linked enzyme aggregates of phenylalanine ammonia lyase (HM-PAL-CLEAs) were developed by co-aggregation of enzyme aggregates with magnetite nanoparticles and subsequent crosslinking with glutaraldehyde. The HM-PAL-CLEAs can be easily separated from the reaction mixture by using an external magnetic field. Analysis by scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM) indicated that PAL-CLEAs were inlayed in nanoparticle aggregates. The HM-PAL-CLEAs revealed a broader limit in optimal pH compared to free enzyme and PAL-CLEAs. Although there is no big difference in Km of enzyme in CLEAs and HM-PAL-CLEAs, Vmax of HM-PAL-CLEAs is about 1.75 times higher than that of CLEAs. Compared with free enzyme and PAL-CLEAs, the HM-PAL-CLEAs also exhibited the highest thermal stability, denaturant stability and storage stability. The HM-PAL-CLEAs retained 30% initial activity even after 11 cycles of reuse, whereas PAL-CLEAs retained 35% of its initial activity only after 7 cycles. These results indicated that hybrid magnetic CLEAs technology might be used as a feasible and efficient solution for improving properties of immobilized enzyme in industrial application.
Research Center for Fermentation Engineering of Hebei, College of Bioscience and Bioengineering, Hebei University of Science and Technology, Shijiazhang, P R China; National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, HaiDian district, Beijing, P R China; Key Laboratory of Industry Microbiology, Ministry of Education, Tianjin University of Science and Technology, Tai Da Development Area, Tianjin, P R China.
Jian dong
Jd
Cui
Research Center for Fermentation Engineering of Hebei, College of Bioscience and Bioengineering, Hebei University of Science and Technology, Shijiazhang, P R China; National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, HaiDian district, Beijing, P R China.
Li li
Ll
Cui
National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, HaiDian district, Beijing, P R China; Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), Tianjin, China.
Song ping
Sp
Zhang
National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, HaiDian district, Beijing, P R China.
Yu fei
Yf
Zhang
National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, HaiDian district, Beijing, P R China.
Zhi guo
Zg
Su
National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, HaiDian district, Beijing, P R China.
Guang hui
Gh
Ma
D001709:Biotechnology / Q000379:methods*; D018592:Cross-Over Studies; D005976:Glutaral / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008280:Magnetics; D058185:Magnetite Nanoparticles / Q000737:chemistry*; D018613:Microscopy, Confocal; D008855:Microscopy, Electron, Scanning; D010650:Phenylalanine Ammonia-Lyase / Q000378:metabolism*; D066329:Protein Aggregates / Q000502:physiology*; D012248:Rhodotorula / Q000201:enzymology* / Q000502:physiology
D058185:Magnetite Nanoparticles; D066329:Protein Aggregates; D010650:Phenylalanine Ammonia-Lyase; D005976:Glutaral