Effective immobilization of lactate dehydrogenase onto mesoporous silica


Alagöz D., Toprak A., Varan N. E. , Yıldırım D., Tükel S. S.

BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY, 2022 (Peer-Reviewed Journal) identifier identifier identifier

  • Publication Type: Article / Article
  • Publication Date: 2022
  • Doi Number: 10.1002/bab.2304
  • Journal Name: BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY
  • Journal Indexes: Science Citation Index Expanded, Scopus, Academic Search Premier, Agricultural & Environmental Science Database, Applied Science & Technology Source, BIOSIS, Biotechnology Research Abstracts, CAB Abstracts, Chemical Abstracts Core, Compendex, Computer & Applied Sciences, EMBASE, Environment Index, Food Science & Technology Abstracts, INSPEC, MEDLINE, Veterinary Science Database
  • Keywords: immobilization, lactate dehydrogenase, mesoporous silica, thermal stabilization, ACID BASED POLYMERS, LACTIC-ACID, COVALENT IMMOBILIZATION, ENZYME IMMOBILIZATION, LIPASE, STABILITY, PROTEINS, SUPPORT, GLUTARALDEHYDE, NANOPARTICLES

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 degrees C for free and 9.0 and 40 degrees C for immobilized l-LDH, respectively. The kinetic parameters (K-m and V-max) 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 degrees C according to calculated t(1/2) values. The immobilized l-LDH retained 80% of its initial activity in a batch reactor after 14 reuses.