Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2022 Dec;69(6):2550-2560.
doi: 10.1002/bab.2304. Epub 2022 Feb 24.

Effective immobilization of lactate dehydrogenase onto mesoporous silica

Dilek Alagöz  1 , Ali Toprak  2 , Nazlı Ece Varan  3 , Deniz Yildirim  4 , S Seyhan Tükel  3
Affiliations

Effective immobilization of lactate dehydrogenase onto mesoporous silica

Dilek Alagöz et al. Biotechnol Appl Biochem. 2022 Dec.

Abstract

This study presents that covalent immobilization technique has been utilized for the immobilization of l-lactate dehydrogenase (l-LDH) from porcine on mesoporous silica. To develop mesoporous silica as support material for use in l-LDH immobilization, the particle surfaces were functionalized with 3-aminopropyltrimethoxysilane and further conjugated with glutaraldehyde. The effect of some parameters such as glutaraldehyde concentration, immobilization pH, initial enzyme concentration, and immobilization time was investigated and the optimum conditions for these parameters were determined as 1% (w/v), pH 8.0, 1 mg/ml, and 120 min, respectively. The maximum working pH and temperature for the oxidation of lactate to pyruvate reaction were determined as 10.0 and 35°C for free and 9.0 and 40°C for immobilized l-LDH, respectively. The kinetic parameters (Km and Vmax ) of l-LDH for the oxidation of lactate to pyruvate reaction were examined as 1.02 mM and 7.58 U/mg protein for free and 0.635 mM and 1.7 U/mg protein for immobilized l-LDH, respectively. Moreover, the immobilized l-LDH was 1.3-fold more stable than free l-LDH at 25°C according to calculated t1/2 values. The immobilized l-LDH retained 80% of its initial activity in a batch reactor after 14 reuses.

Keywords: immobilization; lactate dehydrogenase; mesoporous silica; thermal stabilization.

PubMed Disclaimer

References

REFERENCES

    1. Wang S, Su P, Huang J, Wu J, Yang Yi. Magnetic nanoparticles coated with immobilized alkaline phosphatase for enzymolysis and enzyme inhibition assays. J. Mater. Chem. B. 2013;1:1749-54.
    1. Yildirim D, Alagöz D, Toprak A, Tükel S, Fernandez-Lafuente R. Tuning dimeric formate dehydrogenases reduction/oxidation activities by immobilization. Process Biochem. 2019;85:97-105.
    1. Zahirinejad S, Hemmati R, Homaei A, Dinari A, Hosseinkhani S, Mohammadi S, Vianello F. Nano-organic supports for enzyme immobilization: Scopes and perspectives. Colloids Surf., B. 2021;204: 111774.
    1. Liu D-M, Dong C. Recent advances in nano-carrier immobilized enzymes and their applications. Process Biochem. 2020;92:464-75.
    1. Manoel EA, Pinto M, Dos Santos JCS, Tacias-Pascacio VG, Freire DMG, Pinto JC, Fernandez-Lafuente R. Design of a coreâ “shell support to improve lipase features by immobilization. RSC Adv. 2016;6:62814-24.

LinkOut - more resources