• Home
  • Search Results
  • Recombinant Candida rugosa lipase 2 from Pichia pastoris: immobilization and use as biocatalyst in a stereoselective reaction.

Recombinant Candida rugosa lipase 2 from Pichia pastoris: immobilization and use as biocatalyst in a stereoselective reaction.

Biotechnology progress (2010-10-15)
M Dolors Benaiges, Manuel Alarcón, Pablo Fuciños, Pau Ferrer, Marisa Rua, Francisco Valero

The characterization of the recombinant Candida rugosa Lip2 (r-Lip2) isoenzyme obtained from fed-batch cultures of Pichia pastoris under PAOX promoter was carried out, determining the optimal pH and temperature as well as their catalytic performance in both hydrolysis and synthesis reactions comparing with purified native Lip2 (n-Lip2) previously determined. The substrate specificity of r-Lip2 in hydrolysis reactions was determined with a series of triacylglycerols and p-nitrophenyl esters of variable acyl chain length. r-Lip2 showed the maximum specificity for both substrates towards medium-chain esters (C-8), similar behavior was observed with n-Lip2. However, significant differences were observed towards unsaturated substrates (triolein) or short-chain esters. A statistical design applied to study the effect of pH and temperature on lipase stability shown that r-Lip2, like n-Lip2, was more sensitive to pH than temperature changes. Nevertheless, the overall stability of soluble r-Lip2 was lower than soluble n-Lip2. The stability of r-lip2 was significantly improved by immobilization onto EP100, an excellent support for lipases with yields around 95% for offered lipolytic activity lower than 600 AU/mL. Finally, immobilized r-Lip2 was tested in the resolution of ibuprofen in isooctane by means of enantioselective esterification using 1-butanol as esterifying agent. r-Lip2 showed a better performance in terms of enantiomeric excess (74%) and enatiomeric factor (96%) than n-Lip2 (56 and 80%, respectively) for the same conversion (40%). Thus, r-Lip2 should be considered a good and pure biocatalyst, easy to produce and with a remaining activity of ca. 90% after one reaction cycle when immobilized on EP100.

Product Number
Product Description

2,2,4-Trimethylpentane, anhydrous, 99.8%
2,2,4-Trimethylpentane, analytical standard
2,2,4-Trimethylpentane, suitable for HPLC, ≥99%
2,2,4-Trimethylpentane, HPLC Plus, for HPLC, GC, and residue analysis, ≥99.5%
2,2,4-Trimethylpentane, puriss. p.a., ACS reagent, ≥99.5% (GC)
2,2,4-Trimethylpentane, ReagentPlus®, ≥99%
2,2,4-Trimethylpentane, puriss. p.a., ≥99.5% (GC)
2,2,4-Trimethylpentane, ACS reagent, ≥99.0%
Density Standard 692 kg/m3, H&D Fitzgerald Ltd. Quality