Biosynthesis of raw starch degradingβ-cyclodextrin glycosyltransferase by immobilized cells of Bacillus licheniformis using potato wastewater

The study was sought to enhance the synthesis of thermal stableβ-cyclodextrin glycosyltransferase(β-CGTase)using potato wastewater as a low-cost medium and assess the degree to which it is efficient for industrial production ofβ-cyclodextrin(β-CD)from raw potato starch.Thermophilic bacteria producingβ-CGTase was isolated from Saudi Arabia and the promising strain was identified as Bacillus licheniformis using phylogenetic analysis of the 16S rRNA gene.Alginate-encapsulated cultures exhibited twice-fold ofβ-CGTase production more than free cells.Scanning electron microscopy(SEM)of polymeric capsules indicated the potential for a longer shelf-life,which promotes the restoration of activity in bacterial cells across semi-continuous fermentation ofβ-CGTase production for 252 h.The optimal conditions forβ-CGTase synthesis using potato wastewater medium were at 36 h,pH of 8.0,and 50°C with 0.4%potato starch and 0.6%yeast extract as carbon and nitrogen sources,respectively.The purified enzyme showed a specific activity of 63.90 U/mg with a molecular weight of∼84.6 kDa as determined by SDS-PAGE analysis.The high enzyme activity was observed up to 60°C,and complete stability was achieved at 75°C.High levels of activity and stability were shown at pH 8.0,and the pH range from 7.0–10.0,respectively.The enzyme has an appreciable affinity for raw potato starch with a Km of 5.7×10−6 M and a Vmax of 87.71μmoL/mL/min.β-CD production was effective against 25 U/g of raw potato starch.The outcomes demonstrated its feasibility to develop a fermentation process by integrating the cost-effective production ofβ-CGTase having distinctive properties forβ-CD production with ecofriendly utilization of potato wastewater.

Discovery and Characterization of a Novel Method for Effective Improvement of Cyclodextrin Yield and Product Specificity

Cyclodextrins(CDs) are widely used in food,phamiaceuticals, drug delivery, and chemical industries and in agriculture and environmental engineering. To improve the yield and selectivity of CDs, this work presented a tacile, scalable and efficient enzymatic synthesis of β-CD from starch using β-cyclodextrin glycosyltransferase (CGTase, EC2.4.1.19) from Bacillus cereus. First, we found that the pretreatment of starch dramatically influenced CDs yield that was related to the structure and molecular weight of the substrate starch. Second, alcohol solvents influenced the yield and product selectivity of CDs;tertiary alcohols enhanced CDs yield(from 54.95% to 68.21%) and secondary alcohols increased the product selectivity(β-CD/γ-CD changed from 6.25 to 8.05). Fluorescence quenching analysis showed that the binding constants and entropy of the solvents influenced the yield and product selectivity, respectively. In conclusion, the results demonstrate that this study provides a promising method for the industrial production of β-CD.