TY - JOUR
T1 - A Haloalkane Dehalogenase DhaA Nanoparticle Based on Pullulan Conjugation and Polyethyleneimine Adsorption
AU - Shen, Lijuan
AU - Hu, Lijia
AU - Qi, Jinming
AU - Yu, Weili
AU - Luo, Aiqin
AU - Hu, Tao
N1 - Publisher Copyright:
© The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature 2024.
PY - 2025/3
Y1 - 2025/3
N2 - Haloalkane dehalogenase DhaA is a member of the α/β-hydrolase superfamily and can degrade the halogenated compounds. However, the enzyme could not tolerate harsh and extreme environmental conditions, such as high temperature, extreme pH, and hypersaline, which limits its practical applications. Pullulan is a hydrophilic polysaccharide and acts as an additive to improve the enzyme stability. Polyethyleneimine (PEI) is a protein stabilizer and a polymer with a high density of ionizable amino groups. In the present study, DhaA was covalently conjugated with acetylated pullulan and adsorbed with PEI by electrostatic interactions to form nanoparticles (PEI-pullulan-DhaA). As compared with DhaA, PEI-pullulan-DhaA essentially maintained the enzymatic activity of DhaA, along with slight change in the kinetic parameters and enzyme conformation. The conjugated pullulan tends to form a large hydrated layer around DhaA. PEI, a cationic polymer, generated an amphiphilic microenvironment around DhaA. Pullulan conjugation and PEI adsorption could significantly improve the stability of DhaA against high temperature and low pH by structural stabilization of DhaA. PEI-pullulan-DhaA could also tolerate the hypersaline, organic solvents, and long-term storage. Thus, PEI-pullulan-DhaA has a strong environmental stability and is promising for industrial and environmental applications.
AB - Haloalkane dehalogenase DhaA is a member of the α/β-hydrolase superfamily and can degrade the halogenated compounds. However, the enzyme could not tolerate harsh and extreme environmental conditions, such as high temperature, extreme pH, and hypersaline, which limits its practical applications. Pullulan is a hydrophilic polysaccharide and acts as an additive to improve the enzyme stability. Polyethyleneimine (PEI) is a protein stabilizer and a polymer with a high density of ionizable amino groups. In the present study, DhaA was covalently conjugated with acetylated pullulan and adsorbed with PEI by electrostatic interactions to form nanoparticles (PEI-pullulan-DhaA). As compared with DhaA, PEI-pullulan-DhaA essentially maintained the enzymatic activity of DhaA, along with slight change in the kinetic parameters and enzyme conformation. The conjugated pullulan tends to form a large hydrated layer around DhaA. PEI, a cationic polymer, generated an amphiphilic microenvironment around DhaA. Pullulan conjugation and PEI adsorption could significantly improve the stability of DhaA against high temperature and low pH by structural stabilization of DhaA. PEI-pullulan-DhaA could also tolerate the hypersaline, organic solvents, and long-term storage. Thus, PEI-pullulan-DhaA has a strong environmental stability and is promising for industrial and environmental applications.
KW - DhaA
KW - Enzyme stability
KW - Nanoparticles
KW - Polyethyleneimine
KW - Pullulan
UR - https://www.scopus.com/pages/publications/85210513046
U2 - 10.1007/s12010-024-05117-9
DO - 10.1007/s12010-024-05117-9
M3 - Article
C2 - 39601977
AN - SCOPUS:85210513046
SN - 0273-2289
VL - 197
SP - 1662
EP - 1677
JO - Applied Biochemistry and Biotechnology
JF - Applied Biochemistry and Biotechnology
IS - 3
ER -