Thermal Treatment of Commercial Sweetener Solutions Modulates the Metabolic Responses in C57BL/6 Mice during a 24-Week-Long Exposition

The purpose of this paper was to evaluate the effect of thermal treatment (TT: 121 ± 2 °C, 15 min) on the composition of commercial sweeteners diluted in water (10 °Brix). Additionally, we evaluated the impact of this TT on metabolic responses in C57BL/6 mice during a 24-week treatment. The sweeteners included in this study were sucrose (SC), glucose-63 (GLU63), agave syrup (AS), sucralose (SUC), and steviol glycosides (STG). HPLC analysis showed changes in the concentration of simple sugars of GLU63 and AS after TT. Importantly, in all sweeteners, TT modulated metabolic responses in mice. The mice drinking thermally treated sweetener solution showed an increase of 10–13% (p < 0.05) in food intake (AS, SUC, and STG), beverage intake (2–21%; SC and GLU63), weight gain (38%; SUC), energy (10–13%; AS, SUC, and STG), glucose levels (11–17%; SC and STG), GLP-1 (30%; SC) and insulin (88%; AS) release, and the generation of protein carbonyl (SC) and malondialdehyde (all sweeteners tested) compared to mice drinking solution without TT. In conclusion, TT of sweetener solutions accentuates the metabolic responses of healthy mice, which can be related to overweight and its comorbidities

Cloning, Sequencing, and Expression of the Chitinase Gene chiA74 from Bacillus thuringiensis

The endochitinase gene chiA74 from Bacillus thuringiensis serovar kenyae strain LBIT-82 was cloned in Escherichia coli DH5αF′. A sequence of 676 amino acids was deduced when the gene was completely sequenced. A molecular mass of 74 kDa was estimated for the preprotein, which includes a putative 4-kDa signal sequence located at the N terminus. The deduced amino acid sequence showed high degree of identity with other chitinases such as ChiB from Bacillus cereus (98%) and ChiA71 from Bacillus thuringiensis serovar pakistani (70%). Additionally, ChiA74 showed a modular structure comprised of three domains: a catalytic domain, a fibronectin-like domain, and a chitin-binding domain. All three domains showed conserved sequences when compared to other bacterial chitinase sequences. A ca. 70-kDa mature protein expressed by the cloned gene was detected in zymograms, comigrating with a chitinase produced by the LBIT-82 wild-type strain. ChiA74 is active within a wide pH range (4 to 9), although a bimodal activity was shown at pH 4.79 and 6.34. The optimal temperature was estimated at 57.2°C when tested at pH 6. The potential use of ChiA74 as a synergistic agent, along with the B. thuringiensis insecticidal Cry proteins, is discussed