Influence of maleylation on the physicochemical and functional properties of rapeseed protein isolate was studied. Acylation increased whiteness value and dissociation of proteins, but reduced free sulfhydryl and disulfide content (p < 0.05). Intrinsic fluorescence emission and FTIR spectra revealed distinct perturbations in maleylated proteins' tertiary and secondary conformations. Increase in surface hydrophobicity, foaming capacity, emulsion stability, protein surface load at oil-water interface and decrease in surface tension at air-water interface, occurred till moderate level of modification. While maleylation impaired foam stability, protein solubility and emulsion capacity were markedly ameliorated (p < 0.05), which are concomitant with decreased droplet size distribution (d 32 ). In-vitro digestibility and cytotoxicity tests suggested no severe ill-effects of modified proteins, especially up to low degrees of maleylation. The study shows good potential for maleylated rapeseed proteins as functional food ingredient.

BackgroundTo investigate the potential of Catharanthus roseus leaf aqueous crude extract (CRACE) as a regulator of adipocyte development and function.Methods3T3-L1 adipogenesis model was used to investigate the effect of CRACE on adipogenesis. 3T3-L1 preadipocytes (for adipogenic differentiation) and mature 3T3-L1 adipocytes (for adipocyte function) were treated with non-toxic doses of CRACE. The outcomes were corroborated by intracellular lipid accumulation, expression of pro-and anti-adipogenic effector molecules. To investigate CRACE mediated lipolysis, cAMP accumulation, glycerol release and phosphorylation of key effector molecules were tested in treated mature adipocytes. Finally, the extract was fractionated to identify the active molecule/s in the extract.ResultsCRACE significantly reduced adipocyte differentiation by modulating PPARγ expression. At early stage CRACE directly targeted Lipin1 expression and consequently impacted KLF7, subsequently expression of GATA2, CEBPα, SREBP1c were targeted, with PPARγ expression, particularly curtailed. While CRACE significantly reduced several lipogenic genes like FAS and GPD1 in mature adipocytes, concomitantly, it greatly increased lipolysis resulting in decreased lipid accumulation in mature adipocytes. The increase in lipolysis was due to decreased Akt activation, increased cAMP level, and PKA activity. The fractionation of CRACE allowed identification of two fractions with potent anti-adipogenic activity. Both the fractions contained 1α, 25-dihydroxy Vitamin D3 as major component.Conclusions1α, 25-dihydroxy Vitamin D3 containing CRACE can be developed into an effective anti-obesity formulation that decreases adipogenesis and increases lipid catabolism.

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Use of herbal formulation to modulate adipocyte development and function has been argued as a potent strategy to tackle alarmingly increasing problem of obesity and associated disease like type 2 diabetes and other cardiovascular problems. However, till date no single formulation exists with good efficacy and little side effects. In this study we are investigating the effect of AE11 (6% v/v, nontoxic to 3T3-L1), a plant extract prepared using cold maceration of shade dried leaves of a common flowering plant of Lakhimpur district of Assam, on 3T3-L1 pre-adipocyte differentiation and function. AE11 very efficiently decreased lipid accumulation in differentiating 3T3-L1 cells. To understand the mechanism of such inhibition, we performed gene expression analysis using semi-quantitative PCR for adipogenic master regulator PPAR1 and PPAR2A marked reduction in expression of both of the genes were observed in AE11 treated differentiating 3T3-L1 cells. Western blot analysis confirmed reduction of the two factors at protein level as well. Not surprisingly PPARdownstream GLUT4, PLN1, FABP4, FAS and LPL mRNA content was also reduced in treated groups. Interestingly mRNA content of the transcription factor GATA2, which is a negative regulator of PPAR expression and is normally downregulated during adipogenesis, found to be very high in the AE11 treated cells. This raised a possibility of GATA2 mediated downregulation of PPAR in AE11 treated groups. GATA3 mRNA content was however not different in treated and untreated groups. mRNA of CCAAT enhancer binding protein  (CEBP) which is a positive regulator of PPARexpression was decreased by AE11 exposure to 3T3-L1 cells during differentiation. AE11 targeted the expression of another positive regulator of PPAR expression, SREBP1c. SREBP1c mRNA content was decreased upon AE11 treatment in differentiating 3T3-L1 cells. These preliminary results suggest AE11 is an effective modulator of adipocyte development and function by targeting positive and negative regulators of PPAR gene expression. The authors thank Department of Biotechnology (DBT) for providing fellowship and funds to carry out the work.