Characterization of 36-kDa Dimeric Methotrexate-degrading Enzyme from Variovorax paradoxus, its Performance and Stability of its Cross-linked Aggregates

Document Type

Article

Publication Date

8-1-2023

Abstract

Abstract: In the present study, methotrexate (MTX)-degrading 36-kDa dimer was characterized from the 0–80% ammonium sulphate precipitate of Variovorax paradoxus cell lysate. This enzyme showed optimum activity at 35°C and pH 6.5 with KM value of 199.6 µM for MTX which are different from those of 46-kDa dimer. Micro- or nano-sized cross-linked aggregates of new enzyme showed better stability in the presence of serum than the native soluble form. Addition of an endogenic carrier protein, human serum albumin (HSA) to the enzyme aggregates further improved their serum stability. Nanoaggregates showed better serum stability over microaggregates. Nanoaggregates of the enzyme degraded the MTX faster than soluble form and microaggregates due to their lower K M values on MTX, whereas microaggregates were slower than soluble enzyme itself due to their higher K M values. Hence, cross-linked aggregates of MTX- degrading enzyme isolated from V. paradoxus shown better performance in their nanoform compared to the microform. Nanoaggregates of enzyme revealed the highest functionality and serum stability that makes them more suitable for therapeutic applications.

Identifier

85168369263 (Scopus)

Publication Title

Applied Biochemistry and Microbiology

External Full Text Location

https://doi.org/10.1134/S0003683823040129

e-ISSN

16083024

ISSN

00036838

First Page

468

Last Page

475

Issue

4

Volume

59

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