Bio-specific immobilization of enzymes on electrospun PHB nanofibers.

Enzyme immobilization provides substantial advantages in terms of improving the efficiency of enzymatic process as well as enhancing the reusability of enzymes. Phasins (PhaPs) are naturally occurring polyhydroxyalkanoate (PHA)-binding proteins, and thus can potentially be used as a fusion partner for oriented immobilization of enzymes onto PHA supports. However, presently available granular PHA supports have low surface-area-to-volume ratio and limited configurational flexibility of enzymatic reactions. In this study, we explored the use of electrospun polyhydroxybutyrate (PHB) nanofibers as an alternative support for high density immobilization of a PhaP-fused lipase. As envisioned, the electrospun PHB nanofibers could anchor 120-fold more enzyme than PHB granules of the same weight. Furthermore, the enzymes immobilized onto the PHB nanofibers exhibited markedly higher stability and activity compared to when immobilized on conventional immobilization supports. Our approach combines the advantageous features of nanofibrous material and specificity of biomolecular interaction for the efficient use of enzymes, which can be widely adopted in the development of various enzymatic processes.

High-level production and high-yield recovery of lactobionic acid by the control of pH and temperature in fermentation of Pseudomonas taetrolens.

Lactobionic acid (LBA) was produced by fermentation of Pseudomonas taetrolens. First, to increase the production of LBA by P. taetrolens, we controlled the pH of culture medium by CaCO3 addition (30 g/L) and then examined the initial lactose concentration ranging from 50 to 200 g/L and the growth temperature ranging from 20 to 37 °C. Both the LBA production titer (180 g/L) and the productivity (2.5 g/L h) were highest at 200 g/L lactose concentration and 25 °C of cell growth temperature in shake-flask culture. Although the production of LBA (178 g/L) was almost similar during the batch fermentation of P. taetrolens using 5 L bioreactor, the LBA productivity highly increased to 4.9 g/L h. The method using ethanol precipitation and ion-exchange chromatography was developed to recover the pure LBA from the fermentation broth. The optimum volume of ethanol and pH of culture medium for the precipitation of Ca2+ salt form of LBA were six volume of ethanol and pH 6.5, respectively. The cation-exchange resin T42 finally showed the best recovery yield (97.6%) of LBA from the culture supernatant. The production titer (178 g/L) and the productivity (4.9 g/L h) of lactobionic acid in this study were highest among the previous studies ever reported using P. taetrolens as a production strain of LBA.