TY - JOUR
T1 - Putting precision and elegance in enzyme immobilisation with bio-orthogonal chemistry
AU - Pei, Xiaolin
AU - Luo, Zhiyuan
AU - Qiao, Li
AU - Xiao, Qinjie
AU - Zhang, Pengfei
AU - Wang, Anming
AU - Sheldon, Roger A.
PY - 2022
Y1 - 2022
N2 - The covalent immobilisation of enzymes generally involves the use of highly reactive crosslinkers, such as glutaraldehyde, to couple enzyme molecules to each other or to carriers through, for example, the free amino groups of lysine residues, on the enzyme surface. Unfortunately, such methods suffer from a lack of precision. Random formation of covalent linkages with reactive functional groups in the enzyme leads to disruption of the three dimensional structure and accompanying activity losses. This review focuses on recent advances in the use of bio-orthogonal chemistry in conjunction with rec-DNA to affect highly precise immobilisation of enzymes. In this way, cost-effective combination of production, purification and immobilisation of an enzyme is achieved, in a single unit operation with a high degree of precision. Various bio-orthogonal techniques for putting this precision and elegance into enzyme immobilisation are elaborated. These include, for example, fusing (grafting) peptide or protein tags to the target enzyme that enable its immobilisation in cell lysate or incorporating non-standard amino acids that enable the application of bio-orthogonal chemistry.
AB - The covalent immobilisation of enzymes generally involves the use of highly reactive crosslinkers, such as glutaraldehyde, to couple enzyme molecules to each other or to carriers through, for example, the free amino groups of lysine residues, on the enzyme surface. Unfortunately, such methods suffer from a lack of precision. Random formation of covalent linkages with reactive functional groups in the enzyme leads to disruption of the three dimensional structure and accompanying activity losses. This review focuses on recent advances in the use of bio-orthogonal chemistry in conjunction with rec-DNA to affect highly precise immobilisation of enzymes. In this way, cost-effective combination of production, purification and immobilisation of an enzyme is achieved, in a single unit operation with a high degree of precision. Various bio-orthogonal techniques for putting this precision and elegance into enzyme immobilisation are elaborated. These include, for example, fusing (grafting) peptide or protein tags to the target enzyme that enable its immobilisation in cell lysate or incorporating non-standard amino acids that enable the application of bio-orthogonal chemistry.
UR - http://www.scopus.com/inward/record.url?scp=85135504863&partnerID=8YFLogxK
U2 - 10.1039/d1cs01004b
DO - 10.1039/d1cs01004b
M3 - Review article
AN - SCOPUS:85135504863
SN - 0306-0012
VL - 51
SP - 7281
EP - 7304
JO - Chemical Society Reviews
JF - Chemical Society Reviews
IS - 16
ER -