4.1 Article

Unaided efficient transglutaminase cross-linking of whey proteins strongly impacts the formation and structure of protein alginate particles

Journal

FOOD CHEMISTRY: MOLECULAR SCIENCES
Volume 5, Issue -, Pages -

Publisher

ELSEVIER
DOI: 10.1016/j.fochms.2022.100137

Keywords

LC-MS; MS cross-link identification; Size exclusion chromatography; Dynamic light scattering; Far-and near-UV CD; Intrinsic and ANS fluorescence spectra; Molecular dynamics inter-residue distance analysis

Funding

  1. Novo Nordisk Foundation (NNF) , Denmark Biotechnology Synthesis and Production
  2. Tech-nical University of Denmark
  3. [NNFOC0027616]

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This study challenges the dogma of pretreatment in whey protein modification by showing that enzymatic cross-linking of whey proteins and 13-lactoglobulin can be achieved without pretreatment. The cross-linked proteins had altered structure and showed different complexation behavior with alginate.
There is a dogma within whey protein modification, which dictates the necessity of pretreatment to enzymatic cross-linking of 13-lactoglobulin (13-Lg). Here microbial transglutaminase (MTG) cross-linked whey proteins and 13-Lg effectively in 50 mM NaHCO3, pH 8.5, without pretreatment. Cross-linked 13-Lg spanned 18 to >240 kDa, where 6 of 9 glutamines reacted with 8 of 15 lysines. The initial isopeptide bond formation caused loss of 13-Lg native structure with t1/2 = 3 h, while the polymerization occurred with t1/2 = 10 h. Further, cross-linking effects on protein carbohydrate interaction have been overlooked, leaving a gap in understanding of these complex food matrices. Complexation with alginate showed that 13-Lg cross-linking decreased onset of particle formation, hydrodynamic diameter, stoichiometry (13-Lg/alginate) and dissociation constant. The complexation was favored at higher temperatures (40 degrees C), suggesting that hydrophobic interactions were important. Thus, 13-Lg was cross -linked without pretreatment and the resulting polymers gave rise to altered complexation with alginate.

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