Figure legend
Figure 1 Microscopic images of camelina seeds showing the
effectiveness of Viscozyme® treatment on removing seed
coat mucilage and cotyledon ultrastructure. Upper panels show dissecting
light microscopic images. A dry seed with average length L and width W
indicated (A), a seed showing the hydrated mucilage layer 1 h after
hydration (B), and a dried seed after Viscozyme® treatment (C). Lower
panels show TEM images of a cross-section of cotyledon cells with PSVs
pointed by white arrows and OBs by white arrow heads (D), and OBs at a
higher magnification with black arrows showing the oil body protein
(OBP) layer (E).
Figure 2 Solubility of camelina meal proteins in relation to
pH. Panel A: percentage solubility of the meal protein (%N × 6.25;
one-time extraction) at different pH values. Panel B: polypeptide
profile of soluble proteins at different pH vales. SDS-PAGE separation
was under non-reducing conditions using 8-25% gradient gels.
MWM=Molecular weight markers.
Figure 3 SDS-PAGE of polypeptides from camelina meal, purified
cruciferin and napin. SDS-PAGE profiles of mucilage-reduced, oil-free
meal (A), cruciferin-enriched fraction (B), and napin-enriched fraction
(C) under reducing (+ME) and non-reducing (-ME) conditions. Molecular
weight markers (MWM) are on the left-hand side of each panel with
molecular weights (kDa) of the polypeptide bands shown in right hand
margin of each sample lane in Panel A. Panels D and E show the native
PAGE profile of purified cruciferin and napin, respectively. An 8-25%
gradient gel was used for A, B, D and E, whereas a 20% gel was used for
C. Bovine serum albumin (BSA) and urease were used as standards for
native PAGE.
Figure 4 Two-dimensional electrophoresis separation profiles
and reconstructed images for purified crucifern (A and B) and napin (C
and D) from camelina meal. Numbers in B and D depict the protein spots
in A and C, respectively, that were subjected to LC-MS/MS analysis.
Cartoons in B and D approximate the size and staining intensity observed
in A and C. MWM=Molecular weight markers (kDa).
Figure 5 Polypeptide profiles of OBP from camelina. Panel A:
SDS-PAGE profile in one dimension of meal proteins, intermediate
products (subnatant) and OBP under non-reducing (-ME) and reducing (+ME)
conditions with estimated molecular masses (kDa) of the purified OBPs
obtained using 8-25% gradient precast gels. (MWM=Molecular weight
markers). Panels B and D: Two-dimensional electrophoresis separation of
OBPs on 14% polyacrylamide gels obtained using first dimension IEF pH
3-10 (B) and pH 9-12 (D) followed by SDS-PAGE under non-reducing
conditions for second dimension. Panels C and E: Graphical
representation of B and D, respectively. Numbers in C and E depict the
protein spots observed in A and D, respectively, that were subjected to
LC-MS/MS analysis. Protein spot sizes in C and E approximate the size
and staining intensity observed in B and D. (MWM=Molecular weight
markers).
Figure 6 FTIR spectra of camelina cruciferin and napin together
showing differences in in Amide I region and the regions of phosphorous
containing functional groups. Separate small graphs are for the Fourier
Self Deconvoluted (FSD) spectra of amide I region (1600-1690
cm-1) of cruciferin and napin showing peaks assigned
for secondary structural components. FSD parameters: resolution
enhancement factor (K) = 2.5 (cruciferin) and 2.8 (napin); full width of
half height (FWHH) = 14 cm-1 (cruciferin) and 18
cm-1 (napin); apodization filter = Bassel (cruciferin
and napin).
Figure 7 Changes observed in the far-UV CD spectra of purified
camelina cruciferin (A) and napin (B) with the pH of the medium and the
changes of Fluorescence intensity (C) and the maximum emission
wavelength (λmax) (D) of Tryptophan residues of purified
cruciferin as at ambient temperature (22°C).
Figure 8 Influence of temperature on tryptophan fluorescence of
purified camelina cruciferin at different pH values. Change of
fluorescence intensity (top plot in each panel) and maximum emission
wavelength (λmax) (bottom plot in each panel) at pH 3
(A), pH 7 (B) and pH 10 (C). Values for
F350/F330 ratio are provided in the top
panel.