The nature of the buffer alters the effects of the chemical modification on the stability of immobilized lipases

• Published in: Process biochemistry (1991) Vol. 133; pp. 20 - 27
• Main Authors: Abellanas-Perez, Pedro, Carballares, Diego, Rocha-Martin, Javier, Fernandez-Lafuente, Roberto
• Format: Journal Article
• Language: English
• Published: Elsevier Ltd 01-10-2023
• Subjects: Effect of the buffer nature on enzyme stability; Immobilized lipase stability; Solid phase chemical modification of lipases; Effect of the buffer nature on enzyme stability; Solid phase chemical modification of lipases; Immobilized lipase stability
• ISSN: 1359-5113; 1873-3298
• DOI: 10.1016/j.procbio.2023.08.003

The objective of this paper was to analyze whether an interaction between the effects of the buffer nature and chemical modification on enzyme stability exists. For this, the lipase B from Candida antarctica (CALB) and the lipase from Thermomyces lanuginosus (TLL) were immobilized on octyl agarose beads and modified with picryl sulfonic acid (TNBS) and ethylenediamine via the carbodiimide route. The obtained biocatalysts were them inactivated in different buffers (2-amino-2-(hydroxymethyl)propane-1,3-diol (Tris)-HCl, N-(2-hydroxyethyl)piperazine-N′-(2-ethanesulfonic acid) (HEPES) or phosphate). A significant interaction was found between the chemical modification of the enzyme surfaces and their stabilities in these different buffers. While phosphate was the buffer where the lowest enzyme stabilities were found for both unmodified immobilized enzymes, the differences on enzyme stabilities become much smaller after some chemical modifications. In many instances, chemical modification improves enzyme stability when using a buffer and was negative when using the other one (e.g., TNBS modification of TLL was positive using Tris-HCl or negative using the other buffers, amination of CALB decreased its stability when inactivated in Tris-HCl or HEPES while it almost had no effect in phosphate). Thus, clear co-interactions of the effects on enzyme stability between chemical modification and buffer nature were established.
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•Two lipases were immobilized on octyl-agarose beads exceeding the capacity of the support.•The immobilized enzymes were chemically aminated or hydrophobized.•The inactivation in phosphate, Tris-HCl or HEPES buffers provided very different stabilities.•An interaction buffer nature / chemical modification on enzyme stabilities effects was detected.