Modified silicates and carbon nanotubes for immobilization of lipase from Rhizomucor miehei: Effect of support and immobilization technique on the catalytic performance of the immobilized biocatalysts

ALAGÖZ D., TOPRAK A., YILDIRIM D., TÜKEL S. S., Lafuente R. F.

ENZYME AND MICROBIAL TECHNOLOGY, cilt.144, 2021 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 144
  • Basım Tarihi: 2021
  • Doi Numarası: 10.1016/j.enzmictec.2020.109739
  • Dergi Adı: ENZYME AND MICROBIAL TECHNOLOGY
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus, Academic Search Premier, Artic & Antarctic Regions, BIOSIS, Biotechnology Research Abstracts, CAB Abstracts, Chemical Abstracts Core, Chimica, Compendex, EMBASE, Environment Index, Food Science & Technology Abstracts, INSPEC, MEDLINE, Veterinary Science Database
  • Anahtar Kelimeler: Rhizomucor miehei lipase, Carbon nanotube, Silica, Stabilization, COVALENT IMMOBILIZATION, ENZYME IMMOBILIZATION, PROTEIN IMMOBILIZATION, CHEMICAL-MODIFICATION, IMPROVE BIOCATALYST, MESOPOROUS SILICA, AQUEOUS-SOLUTION, AGAROSE BEADS, CROSS-LINKER, SPACER-ARM
  • Çukurova Üniversitesi Adresli: Evet

Özet

Lipase from Rhizomucor miehei (RML) was covalently immobilized on different supports, two silica gels and two carbon nanotube samples, using two different strategies. RML was immobilized on 3-carboxypropyl silica gel (RML@Si-COOH) and multi-wall carbon nanotubes containing carboxylic acid functionalities (RML@MCNT-COOH) using a two-step carbodiimide activation/immobilization reaction. Moreover, the enzyme was also immobilized on 3-aminopropyl silica (RML@Si-Glu) and single-wall carbon nanotubes functionalized with 3-APTES and activated with glutaraldehyde (RML@SCNT-Glu). Before and after RML immobilization, the structurel properties of supports were characterized and compared in detail. After immobilization, the expressed activities were 36.9, 90.2, 16.9, and 26.1 % for RML@Si-COOH, RML@Si-Glu, RML@MCNT-COOH, and RML@SCNT-Glu, respectively. The kinetic parameters of free and immobilized RML samples were determined for three substrates, p-nitrophenyl acetate, p-nitrophenyl butyrate and p-nitrophenyl palmitate, and RML@Si-Glu showed higher catalytic efficiency than the other immobilized RML samples. RML@Si-COOH, RML@Si-Glu, RML@MCNT-COOH, and RML@SCNT-Glu exhibited 5.8, 7.6, 4.2 and 4.6 folds longer half-life values than those of the free enzyme at pH 7.5 and 40 degrees C. Recyclability studies showed that all the immobilized RML biocatalysts retained over 90 % of their initial activities after ten cycles in the hydrolysis of p-nitrophenyl butyrate.