Upscale of recombinant a-L-rhamnosidase production by XXXX Mut(S) strain.

@entity10056 @entity1042 @entity26 @entity653 @entity16956
@entity10056 is currently one of the most preferred microorganisms for recombinant enzyme production due to its efficient expression system. The advantages include the production of high amounts of recombinant proteins containing the appropriate posttranslational modifications and easy cultivation conditions. a-L-Rhamnosidase is a biotechnologically important enzyme in food and pharmaceutical industry, used for example in debittering of citrus fruit juices, rhamnose pruning from @entity16956 , or enhancement of wine aromas, creating a demand for the production of an active and stable enzyme. The production of recombinant a-L-rhamnosidase cloned in the Mut(S) strain of @entity10056 KM71H was optimized. The encoding gene is located under the control of the AOX promoter, which is induced by @entity1042 whose concentration is instrumental for these strain types. Fermentation was upscaled in bioreactors employing various media and several @entity1042 -feeding strategies. It was found that fed batch with BSM media was more effective compared to BMMH (Buffered @entity1042 -complex Medium) media due to lower cost and improved biomass formation. In BSM (Basal Salt Medium) medium, the dry cell weight reached approximately 60 g/L, while in BMMH it was only 8.3 g/L, without additional @entity653 , which positively influenced the amount of enzyme produced. New @entity1042 feeding strategy, based on the level of dissolved @entity26 was developed in this study. This protocol that is entirely independent on @entity1042 monitoring was up scaled to a 19.5-L fermenter with 10-L working volume with the productivity of 13.34 mgprot/L/h and specific activity of a-L-rhamnosidase of 82 U/mg. The simplified fermentation protocol was developed for easy and effective fermentation of @entity10056 Mut(S) based on dissolved @entity26 monitoring in the induction phase of an enzyme production.
