| Products/Services Used | Details | Operation |
|---|---|---|
| Codon Optimization | Two different plasmids were used, namely: the pPICZαA plasmid containing Rhizopus oryzae lipase (rROL) as expressed by the inducible promoter alcohol oxidase 1 (PAOX1), which was labeled “rROL plasmid”, and a derivative thereof containing the last 28 amino acids of the prosequence in the N-terminal of rROL (proROL). The region corresponding to the 28 amino acids was codon-optimized for P. pastoris (GenScript, Piscataway, NJ, USA) and flanked with XhoI and BtsαI restriction sites for subsequent digestion-ligation cloning. The rROL plasmid natively contained both restriction sites, so cloning left no unwanted sequences in it. The new plasmid was labeled “proROL plasmid”. The Supplementary Information includes the plasmids (Supplementary Materials File S1). | Get A Quote |
ecombinant Rhizopus oryzae lipase (mature sequence, rROL) was modified by adding to its N-terminal 28 additional amino acids from the C-terminal of the prosequence (proROL) to obtain a biocatalyst more suitable for the biodiesel industry. Both enzymes were expressed in Pichia pastoris and compared in terms of production bioprocess parameters, biochemical properties, and stability. Growth kinetics, production, and yields were better for proROL harboring strain than rROL one in batch cultures. When different fed-batch strategies were applied, lipase production and volumetric productivity of proROL-strain were always higher (5.4 and 4.4-fold, respectively) in the best case. rROL and proROL enzymatic activity w... More
