Engineering pichiapastorisExpression system for enhancedCandida antarcticalipase b Enzyme secretion

Abstract

Candida antarcticalipases have gained significant commercial newlineimportance and as the expression levels in the native organism are too low newlinethere is a need for recombinant overexpression for large scale production of newlinethis biocatalyst In the production of recombinant proteins yeasts offer newlineadvantages over both bacterial and mammalian systems In eukaryotes yeasts newlineare easy to manipulate have a relatively rapid growth rate and can be grown newlineto higher cell densities than bacteria On the other hand as they are newlineeukaryotic they offer the capacity to posttranslationally modify the newlinetranslated protein and are able to secrete proteins in a simple medium at a newlinefraction of the cost of a mammalian expression system The methylotropic newlineyeastPpastorishas gained much popularity in recent years due to its newlineadvantages such as easy manipulation at the molecular genetic level newlineexpression of proteins at very high level intracellularly or extracellularly and newlineits growth in simple and cheap medium to very high cell densities Hence an newlineattempt was made to produce the Candida antarcticalipase CalB enzyme in newlineP pastoris newlineIn Pichiapastoris heterologous protein secretion is limited due to newlinesuboptimal folding conditions inside the cell The Hsp70 and Hsp40 newlinechaperone families in the cytoplasm or in endoplasmic reticulum ER newlineimportantly regulate the translocation folding and secretion of heterologous newlineproteins newline newline