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Codon Optimization | … The laccase coding sequence (GenBank HQ857207) was codon optimized for expression in E. coli (European Nucleotide Archive LT795002), synthesized by Genscript (Piscataway, USA) and transferred to the expression vector pET-22b(+) (Novagen, Darmstadt, Germany) … | Get A Quote |
Laccases are used for the conversion of biomass into fermentable sugars but it is difficult to produce high yields of active laccases in heterologous expression systems. We overcame this challenge by expressing Streptomyces cyaneus CECT 3335 laccase in Escherichia coli (ScLac) and we achieved a yield of up to 104?mg?L following purification by one-step affinity chromatography. Stability and activity assays using simple lignin model substrates showed that the purified enzyme preparation was active over a broad pH range and at high temperatures, suggesting it would be suitable for biomass degradation. The reusability of ScLac was also demonstrated by immobilizing the enzyme on agarose beads with a binding yield... More