Browsing by Subject "Enzymes"

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Importance of the 5’ untranslated region for recombinant enzyme production in isolated Bacillus subtilis 007
(2025) Senger, Jana; Schulz, Adriana; Seitl, Ines; Heider, Martin; Fischer, Lutz; Senger, Jana; Institute of Food Science and Biotechnology, Department of Biotechnology and Enzyme Science, University of Hohenheim, Garbenstr. 25, 70599, Stuttgart, Germany; Schulz, Adriana; Institute of Food Science and Biotechnology, Department of Biotechnology and Enzyme Science, University of Hohenheim, Garbenstr. 25, 70599, Stuttgart, Germany; Seitl, Ines; Institute of Food Science and Biotechnology, Department of Biotechnology and Enzyme Science, University of Hohenheim, Garbenstr. 25, 70599, Stuttgart, Germany; Heider, Martin; Institute of Food Science and Biotechnology, Department of Biotechnology and Enzyme Science, University of Hohenheim, Garbenstr. 25, 70599, Stuttgart, Germany; Fischer, Lutz; Institute of Food Science and Biotechnology, Department of Biotechnology and Enzyme Science, University of Hohenheim, Garbenstr. 25, 70599, Stuttgart, Germany
The production of industrial enzymes requires an efficient expression system with a suitable host. This study investigated the isolated Bacillus subtilis 007 as a host for expressing three enzymes with potential application in the food industry. Firstly, testing the PaprE and P43 promoters and the corresponding 5’ untranslated regions revealed great differences in the production of the recently discovered β-galactosidase from Paenibacillus wnnyii. Expression controlled by the PaprE promoter yielded a significantly higher activity of 2515 µkat/L, compared to 56 µkat/L with the P43 promoter. Modifications on the PaprE core promoter region or the spacer, the sequence between the Shine-Dalgarno sequence and the start codon, did not improve β-galactosidase production. Since the aprE 5’ untranslated region contributes to a high mRNA stability, it was incorporated into the P43 construct to determine whether mRNA stability is responsible for the differences observed in β-galactosidase production. Interestingly, mRNA stability was significantly improved and led to a nearly 50-fold higher β-galactosidase production of 2756 µkat/L. This strategy was successfully validated by the expression of two other enzymes: the cellobiose-2-epimerase from Caldicellulosiruptor saccharolyticus and the β-glucosidase from Pyrococcus furiosus. These findings underscored the crucial role of post-transcriptional regulation and emphasized mRNA stability as a key role in recombinant enzyme production in B. subtilis 007.
Multi‐scale dough adhesion analysis: Relation between laboratory scale, pilot scale and human sensory
(2023) Vogt, Ulrike Therese; Kwak, Ju Eun; Fahmy, Ahmed Raouf; Laukemper, Rita; Henrich, Alexander; Becker, Thomas; Jekle, Mario
Undesired dough adhesion is still a challenge during the production of baked goods. There are various methods for determining the adhesive texture properties of dough. In the majority of scientific papers, dough stickiness is measured analytically by the force‐distance recording of dough detachment. In this study, we describe a new multi‐scale approach to compare dough adhesion phenomena in a laboratory, pilot sale and human sensory assessment. In it, the adhesive material properties of dough were investigated using a pilot scale toppling device representing dough adhesion behavior in the production process, in the laboratory by texture analysis with the Chen–Hoseney method and furthermore with a new, implemented non‐oral human sensory analysis. To simulate different dough adhesion behavior, the dough mechanical and adhesion properties were varied by applying dough‐modifying enzymes and different dough storage times. The structural changes in the different wheat dough system were compared by rheological characterization. By characterizing the different adhesion phenomena of the doughs, the sample with bacterial xylanase showed the highest values after 80 min of storage time in all three methods. Correlation analysis revealed a strong relationship between the detachment time (pilot scale) and human sensory assessment attributes (Force R = 0.81, Time R = 0.87, Distance R = 0.92, Stickiness R = 0.80) after 80 min of storage time. Even though human sensory assessment showed limits in the detectability of differences in dough adhesion behavior compared to the Chen–Hoseney method, it was better suited to predict machinability.