TY - JOUR
T1 - Precision fermentation as a route to modify β-lactoglobulin structure through substitution of specific cysteine residues
AU - Brune, Sarah E.
AU - Hoppenreijs, Loes J.G.
AU - Kühl, Toni
AU - Lautenbach, Vanessa
AU - Walter, Johannes
AU - Peukert, Wolfgang
AU - Schwarz, Karin
AU - Imhof, Diana
AU - Boom, Remko M.
AU - Krull, Rainer
AU - Keppler, Julia K.
AU - Biedendieck, Rebekka
PY - 2023/12
Y1 - 2023/12
N2 - Precision fermentation is a promising route to develop recombinant proteins with improved functionality. Protein functionality is highly influenced by protein structure, which can be significantly affected by conformational restraints induced by cysteines. Here, we study the impact of individual cysteines on β-lactoglobulin structure, by stepwise substituting them with alanine. Structural characterisation was performed from secondary to quaternary level. The findings demonstrated that the free Cys121 played no role in the folding and dimerisation of BLG. The β-barrel usually includes one disulphide bond (Cys106–Cys119), which was found to be crucial for its formation. The usually exposed disulphide bond (Cys66–Cys160) played no major role in folding of the β-barrel, but mainly modulated its accessibility and mediated dimerisation. Thereby, cysteine mutations can be used as a tool to modify protein structures. These different structures can now be used to further understand the impact on protein behaviour and their functionality.
AB - Precision fermentation is a promising route to develop recombinant proteins with improved functionality. Protein functionality is highly influenced by protein structure, which can be significantly affected by conformational restraints induced by cysteines. Here, we study the impact of individual cysteines on β-lactoglobulin structure, by stepwise substituting them with alanine. Structural characterisation was performed from secondary to quaternary level. The findings demonstrated that the free Cys121 played no role in the folding and dimerisation of BLG. The β-barrel usually includes one disulphide bond (Cys106–Cys119), which was found to be crucial for its formation. The usually exposed disulphide bond (Cys66–Cys160) played no major role in folding of the β-barrel, but mainly modulated its accessibility and mediated dimerisation. Thereby, cysteine mutations can be used as a tool to modify protein structures. These different structures can now be used to further understand the impact on protein behaviour and their functionality.
U2 - 10.1016/j.idairyj.2023.105772
DO - 10.1016/j.idairyj.2023.105772
M3 - Article
AN - SCOPUS:85169910361
SN - 0958-6946
VL - 147
JO - International Dairy Journal
JF - International Dairy Journal
M1 - 105772
ER -