Creating an Efficient Methanol-Stable Biocatalyst by Protein and Immobilization Engineering Steps towards Efficient Biosynthesis of Biodiesel

Shalev Gihaz, Diána Weiser, Adi Dror, Péter Sátorhelyi, Moran Jerabek-Willemsen, László Poppe, Ayelet Fishman

Research output: Contribution to journalArticlepeer-review

Abstract

Two ternary sol–gel matrices, an octyltriethoxysilane-based aliphatic matrix and a phenyltriethoxysilane (PTEOS)-based aromatic matrix, were used to immobilize a methanol-stable variant of lipase from Geobacillus stearothermophilus T6 for the synthesis of biodiesel from waste oil. Superior thermal stability of the mutant versus the wildtype in methanol was confirmed by intrinsic protein fluorescence measurements. The influence of skim milk and soluble E. coli lysate proteins as bulking and stabilizing agents in conjunction with sol–gel entrapment were investigated. E. coli lysate proteins were better stabilizing agents of the purified lipase mutant than skim milk, as evidenced by reverse engineering of the aromatic-based system. This was also shown for commercial Candida antarctica lipase B (CaLB) and Thermomyces lanuginosus lipase (TLL). Uniform, dense, and nonaggregated particles imaged by scanning electron microscopy and a small particle size of 13 μm pertaining to the system comprising PTEOS and E. coli lysate proteins correlated well with high esterification activity. Combining protein and immobilization engineering resulted in a durable biocatalyst with efficient recycling ability and high biodiesel conversion rates.

Original languageEnglish
Pages (from-to)3161-3170
Number of pages10
JournalChemSusChem
Volume9
Issue number22
DOIs
StatePublished - 23 Nov 2016

Keywords

  • biodiesel
  • enzymes
  • immobilization
  • protein engineering
  • sol–gel processes

All Science Journal Classification (ASJC) codes

  • General Chemical Engineering
  • General Energy
  • General Materials Science
  • Environmental Chemistry

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