1. There are different isoforms of this enzyme in swiss prot, would you mind please help me to find out which isoform is better for expression in Pichia pastoris?
2. I want to use recombinant laccase for bioremediation of wastewater. Which isoform do you recommend for this purpose?
3. I want to do codon optimization based on the synonymous codon bias of P.pastoris and optimize G+C content for further improvement of the expression level of recombinant laccase. Would you mind please tell me which software and company should I use?
4. Can we use testament enzyme for laccase isolated from other different sources? (For example laccase from Pleurotus ostreatus (mushroom) sigma 75117)
Hi! Regarding Point 3: I normally use GeneArt or DNA 2.0 for codon optimization for Pichia. DNA 2.0 is a bit more expensive but they claim to have the best algorithm for codon optimization, therefore I prefer it for Pichia.
There are too many open questions and I would recommend to do an extensive literature search first.
There are several organism with several laccase isoenzymes you might start with. You should try to find out which enzyme will have best activity for your bioremediation under process conditions and then start to clone and express the enzyme.
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