Trehalose, a naturally occurring osmolyte, is considered as a universal protein stabilizer. We investigated the effect of the disaccharides, trehalose and sucrose, on the thermal stability and conformation of bromelain. To our surprise, bromelain in the presence of 1 M trehalose/sucrose was destabilized under thermal stress. The average Tm values as determined by UV spectroscopy and CD spectropolarimetry decreased by 5 degrees and 7 degrees C for bromelain in 1 M sucrose or trehalose solutions, respectively. The enzyme was also found to inactivate faster at 60 degrees C in the presence of these osmolytes. The tertiary and secondary structure of bromelain undergoes small changes in the presence of sucrose/trehalose. Studies on the binding of these osmolytes with the native and the heat denatured enzyme revealed that sucrose/trehalose lead to preferential hydration of the denatured bromelain as compared to the native one, hence stabilizing more the denatured conformation. This is perhaps the first report on the destabilization of a protein by trehalose. Study holds ProTherm entries: 23984, 23985, 23986, 23987, 23988 Extra Details: destabilization; stem bromelain; temperature; trehalose
Submitter: Connie Wang
Submission Date: April 24, 2018, 8:55 p.m.
|Number of data points||5|
|Proteins||Stem bromelain ; STEM BROMELAIN|
|Assays/Quantities/Protocols||Experimental Assay: Tm prot_conc:0.5 mg/mL, details:Additives 1M trehalose ; Experimental Assay: Tm prot_conc:0.5 mg/mL, details:Additives ; Experimental Assay: Tm prot_conc:1 mg/mL, details:Additives 1M trehalose ; Experimental Assay: Tm prot_conc:1 mg/mL, details:Additives sucrose (1M), ; Experimental Assay: Tm prot_conc:1 mg/mL, details:Additives|
|Libraries||Mutations for sequence VPQSIDWRDYGAVTSVKNQNPCGACWAFAAIATVESIYKIKKGILEPLSEQQVLDCAKGYGCKGGWEFRAFEFIISNKGVASGAIYPYKAAKGTCKTDGVPNSAYITGYARVPRNNESSMMYAVSKQPITVAVDANANFQYYKSGVFNGPCGTSLNHAVTAIGYGQDSIIYPKKWGAKWGEAGYIRMARDVSSSSGICGIAIDPLYPTLE|
|Percent Identity||Matching Chains||Protein||Accession||Entry Name|