Hexafluoroisopropanol-induced helix–sheet transition of stem bromelain: Correlation to function

Inhibition of Adipogenesis and Induction of Apoptosis and Lipolysis by Stem Bromelain in 3T3-L1 Adipocytes

“…SBM solutions of native and catalytic-ally inactive were prepared, dialyzed extensively and subjected to size exclusion chromatography as mentioned elsewhere [28], [42], [43]. MTT assay reagents, Oil Red O, haematoxylin stain, insulin, β-nicotinamide adenine dinucleotide, dihydroxyacetone-3-phosphate, triethanolamine, atRA (purity ≥98% (HPLC)), 1,25-dihydroxyvitamin D3 (1,25(OH) 2 D 3 ; vitamin D) (purity ≥99% (HPLC)), dexamethasone, and 3-isobutyl-1-methylxanthine were obtained from Calbiochem.…”

“…In addition, it is being used efficaciously as an anti-obesity pill (France Vita Natura Pvt Ltd.). While its anti-metastatic effects and its modulation of immune cells have been adequately addressed [25], [27], [28], [29], its mechanism of action on adipocytes, however has not been addressed or understood. The diverse effects of SBM appear to depend on its unusual ability to traverse membranes and its acceptability as an orally administered enzyme with minimal side effects [24], [30], [31], [32], [33].…”

Trifluoroethanol stabilizes the molten globule state and induces non-amyloidic turbidity in stem bromelain near its isoelectric point

“…Although no data are available on its crystal structure, the amino acid sequence of SBM is similar to that of papain [55], actinidin [56], proteinase [57], and chymopapain [58]. SBM is unique in its therapeutic benefits and in some of its folding behaviors in response to chaotropic agents [22,50]. Thus, despite the sequence similarities, these proteins might not share common folding patterns and could have different structures.…”

“…3B) [45]. SBM contains five tryptophan residues: three are buried in the hydrophobic core and two are located near the surface of the molecule [50]. Comparisons of the effective quenching constant (K c ) and the fraction of accessible fluorophore (f a ) provide information about the changes in the accessibility of tryptophanyl residues of SBM at pH 10 and TFE concentrations of 0-40% ( Table 1).…”

Characteristics of thermostable amylopullulanase of Geobacillus thermoleovorans and its truncated variants

“…ANS is known to bind the hydrophobic regions of the protein and produces fluorescence in the wavelength range 400-600 nm with the emission maximum occurring at 470-490 nm [38][39][40][41]. It has, therefore, been successfully used for monitoring structural changes in proteins during both denaturation and renaturation processes [42,43].…”