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Nano Letters Feb 2020Mucus is a viscoelastic biological hydrogel that protects the epithelial surface from penetration by most nanoparticles, which limits the efficiency of oral drug...
Mucus is a viscoelastic biological hydrogel that protects the epithelial surface from penetration by most nanoparticles, which limits the efficiency of oral drug delivery. Pursuing highly efficient, biocompatible, and biodegradable oral drug vehicles is of central importance to the development of promising nanomedicine. Here, we prepared five peptosomes (PSs) with various sizes, shapes, and rigidities based on self-assembly of amphiphilic α-lactalbumin (α-lac) peptides from partial enzymolysis and cross-linking. The mucus permeation of α-lac PSs and release of curcumin (Cur) encapsulated in these PSs were evaluated. Compared with a long nanotube, big nanosphere, small nanosphere, and cross-linked short nanotube, we demonstrated that a short nanotube (SNT) exhibits excellent permeability in mucus, which enables it to arrive at epithelial cells quickly. Besides, SNT exhibits the highest cellular uptake and transmembrane permeability on Caco-2/HT29-MTX (E12) 3D coculture model. In vivo pharmacokinetic evaluation revealed that SNT formulation shows the highest curcumin bioavailability, which is 6.85-folds higher than free Cur. Most importantly, Cur loaded in SNT exhibits the optimum therapeutic efficacy for in vivo treatment of dextran sulfate sodium (DSS)-induced ulcerative colitis. In the end, the mechanism of the high permeability of SNTs through mucus was explained by coarse-grained molecular dynamics simulations, which indicated that short time scale jiggling and flying across pores of mucus network played key roles. These findings revealed the tubular α-lac PSs could be a promising oral drug delivery system targeted to mucosal for improving absorption and bioavailability of hydrophobic bioactive ingredients.
Topics: Animals; Biological Availability; Caco-2 Cells; Colitis, Ulcerative; Curcumin; Dextran Sulfate; Drug Carriers; Humans; Intestines; Lactalbumin; Mice; Mucus; Nanoparticles; Nanospheres; Nanotubes; Permeability
PubMed: 31904988
DOI: 10.1021/acs.nanolett.9b04841 -
Journal of Agricultural and Food... Aug 2021Improving the stability and bioavailability of catechins is of great importance. Epigallocatechin (EGC), the major catechin in green tea, is a potent antioxidant with...
Improving the stability and bioavailability of catechins is of great importance. Epigallocatechin (EGC), the major catechin in green tea, is a potent antioxidant with numerous attributed health benefits. However, the low permeability and stability limit its enrichment in the diet for preventive medicine. In this study, we explored the interaction of EGC and α-lactalbumin by spectroscopic, thermodynamic, and crystallographic methods. The isothermal titration calorimetry experiments elucidated that α-lactalbumin binds to EGC at a ratio of 1:1 with a low affinity of (4.01 ± 0.11) × 10 M. A crystal structure solved at a high resolution (1.2 Å) provided direct evidence for the weak interaction between EGC and α-lactalbumin at an atomic level. The novel binding site was discovered at the exterior surface of α-lactalbumin for the first time, supporting a new binding behavior. Consequently, our results demonstrated that the binding of α-lactalbumin to EGC could protect EGC against light-induced, thermal-induced, and pH-induced damage. More importantly, the formed complex has better bioaccessibility than unbound EGC, which was approved by a cell absorption experiment. Such research is beneficial for designing protein-based nanocarriers for polyphenols.
Topics: Caco-2 Cells; Catechin; Humans; Lactalbumin; Tea
PubMed: 34286590
DOI: 10.1021/acs.jafc.1c03427 -
Food Chemistry Aug 2023Alpha-lactalbumin (α-La) is a crucial active component in whey protein. It would be mixed with edible azo pigments during processing. Spectroscopic analyses and...
Alpha-lactalbumin (α-La) is a crucial active component in whey protein. It would be mixed with edible azo pigments during processing. Spectroscopic analyses and computer simulations were used here to characterize the interaction between acid red 27 (C27) /acidic red B (FB) and α-La. Fluorescence, thermodynamics, and energy transfer showed the binding mechanism is a static quenching with a medium affinity. This binding process occurred spontaneously and was mainly driven by hydrophobic forces. Conformation analysis showed FB led to a greater change in the secondary structure of α-La compared with C27. C27 increased and FB decreased the surface hydrophobicity of α-La. The spatial structures of complexes were visualized with computer aid. The azo colorant binds to α-La easily and deeply with a smaller space volume and dipole moment and thereby affecting the α-La conformation and functionality. This study provides a theoretical basis for the application of edible azo pigments.
Topics: Lactalbumin; Whey Proteins; Hydrophobic and Hydrophilic Interactions; Protein Structure, Secondary
PubMed: 36893641
DOI: 10.1016/j.foodchem.2023.135826 -
Applied Spectroscopy Jun 2021Characterization and quantification of individual whey proteins are of crucial importance to many industrial dairy processes. Labor intensive wet-chemical methods are...
Characterization and quantification of individual whey proteins are of crucial importance to many industrial dairy processes. Labor intensive wet-chemical methods are still being used for this purpose, but a rapid quantification method for individual whey proteins is highly desired. This work investigate the utility of Fourier transform mid-infrared spectroscopy and Fourier transform near-infrared spectroscopy for rapid quantification of the two main whey proteins (β-lactoglobulin and α-lactalbumin) in complex aqueous whey solutions simulating production process streams. MIR and NIR spectra obtained on whey samples with known and varying amounts of the proteins of interest and are used to develop partial least squares prediction models. Selection of informative wavelength regions allowed for prediction of β-lactoglobulin and α-lactalbumin concentrations with very high precision and accuracy (root mean square error of cross-validation, or RMSECV, of 0.6% and R of 0.99 for NIR), demonstrating the potential of being implemented for rapid in-line monitoring of protein composition in industrial whey streams. Two-dimensional MIR-NIR correlation spectroscopy is used to identify the most informative parts of the NIR spectra in relation to protein secondary structure. In addition multivariate curve resolution is applied to the MIR data to resolve mixture spectra and to elucidate the spectral ranges that were most useful in distinguishing between the two whey proteins. The study concludes that NIR spectroscopy has potential for accurate in-line protein quantification and overall secondary protein structure quantification which open new possibilities for in-line industrial applications.
Topics: Fourier Analysis; Lactalbumin; Lactoglobulins; Least-Squares Analysis; Spectroscopy, Fourier Transform Infrared; Spectroscopy, Near-Infrared; Whey; Whey Proteins
PubMed: 33231482
DOI: 10.1177/0003702820979747 -
Scientific Reports Apr 2021To verify whether myo-inositol plus α-lactalbumin may reduce insulin resistance and excessive fetal growth in women with gestational diabetes mellitus. In a 12-month... (Randomized Controlled Trial)
Randomized Controlled Trial
To verify whether myo-inositol plus α-lactalbumin may reduce insulin resistance and excessive fetal growth in women with gestational diabetes mellitus. In a 12-month period, 120 women with a diagnosis of gestational diabetes mellitus were consecutively enrolled with an allocation of 1:1 in each group and randomly treated with myo-inositol plus α-lactalbumin plus folic acid (treated group) or folic acid (control group) for 2 months. Primary outcome was the variation of insulin resistance through the study evaluated by HOMA-IR. Secondary outcome was the evaluation, through the study, of fetal growth by ultrasound measurements of abdominal circumference centiles and estimated fat thickness. Some clinical outcomes were also considered. After 2 months, in the treated group, a significant reduction in insulin resistance (HOMA values 3.1 ± 1.4 vs 6.1 ± 3.4, p = 0.0002) and fetal growth was shown (Abdominal circumference centiles 54.9 ± 23.5 vs 67.5 ± 22.6, P = 0.006). Among clinical outcomes, a significant decrease in the rate of women who needed insulin (6.7% vs 20.3%, p = 0.03) and of pre-term birth (0 vs 15.2%, p = 0.007) was evidenced. A combination of myo-inositol and α-lactalbumin may reduce insulin resistance and excessive fetal growth.Clinical trial registration: ClinicalTrials.gov, http://www.clinicaltrials.gov , NCT03763669, first posted date 04/12/2018; last posted date December 06/12/2018.
Topics: Adult; Birth Weight; Case-Control Studies; Diabetes, Gestational; Dietary Supplements; Female; Folic Acid; Humans; Inositol; Insulin Resistance; Lactalbumin; Pregnancy; Pregnancy Outcome
PubMed: 33893377
DOI: 10.1038/s41598-021-88329-x -
Plant Cell Reports Oct 2022This study demonstrated high expression and accumulation of human α-lactalbumin in transgenic maize, and significant improvement of lysine content in maize endosperm....
This study demonstrated high expression and accumulation of human α-lactalbumin in transgenic maize, and significant improvement of lysine content in maize endosperm. As a high-yield crop, lack of lysine in endosperm storage protein is a major defect of maize (Zea mays L.). Specifically expression of foreign proteins is a potential way to improve lysine content in maize endosperm. Human α-lactalbumin is such a protein with high lysine content and high nutritional value. In this study, the codon-optimized human lactalbumin alpha (LALBA) gene was driven by maize endosperm-specific 27 kD γ-zein promoter, and transformed into maize. Five independent transgenic lines were obtained, and LALBA was highly expressed in endosperm in all these lines. Protein assay indicated that human α-lactalbumin was highly accumulated in maize endosperm. Immuno-localization assay indicated that human α-lactalbumin was mainly deposited into the protein body (PB). Protein interaction assay showed that human α-lactalbumin interacted with 16 kD γ-zein, which might lead to its deposition to the PBs. Amino acid analysis of two independent transgenic lines showed significant increase of lysine contents in transgenic endosperm, with 47.26% and 45.15% increase to their non-transgenic seeds, respectively. We obtained transgenic maize with endosperm-specific accumulation of human α-lactalbumin at high level and increased the lysine content in maize endosperm. This study demonstrated an effective way to improve the nutritional value of maize seeds.
Topics: Amino Acids; Codon; Endosperm; Humans; Lactalbumin; Lysine; Plants, Genetically Modified; Seeds; Transcription Factors; Zea mays; Zein
PubMed: 35918456
DOI: 10.1007/s00299-022-02906-6 -
Cancer Investigation Oct 2017Alpha-lactalbumin (α-LA), a small milk calcium-binding globular protein, is known to possess noticeable anticancer activity, which is determined by the ability of this... (Comparative Study)
Comparative Study
Alpha-lactalbumin (α-LA), a small milk calcium-binding globular protein, is known to possess noticeable anticancer activity, which is determined by the ability of this protein to form complexes with oleic acid (OA). To date, in addition to human and bovine α-LA, the ability to form such anti-tumor complexes with OA was described for goat and camel α-LA. Although the mechanisms of the anticancer activity of human and bovine α-LA are already well-studied, little is currently known about the anticancer action of this camel protein. The goal of this study was to fill this gap and to analyze the anticancer and pro-apoptotic activities of camel α-LA in its free form (α-cLA) and as an OA-containing complex (OA-α-cLA) using four human cancer cell lines, including Caco-2 colon cancer cells, PC-3 prostate cancer cells, HepG-2 hepatoma cells, and MCF-7 breast cancer cells as targets. The anti-tumor activities of OA-α-cLA and α-cLA were analyzed using MTT test, annexin/PI staining, cell cycle analysis, nuclear staining, and tyrosine kinase (TK) inhibition methods. We show here that the OA-α-cLA complex does not affect normal cells but has noticeable anti-cancer activity, especially against MCF-7 cells, thus boosting the anticancer activity of α-cLA and improving the selectivity of OA. The OA-α-cLA complex mediated cancer cell death via selective induction of apoptosis and cell-cycle arrest at lower IC than that of free α-cLA by more than two folds. However, OA induced apoptosis at higher extent than OA-α-cLA and α-cLA. OA also caused unselective apoptosis-dependent cell death in both normal and cancer cells to a similar degree. The apoptosis and cell-cycle arresting effect of OA-α-cLA may be attributed to the TK inhibition activity of OA. Therefore, OA-α-cLA serves as efficient anticancer complex with two functional components, α-cLA and OA, possessing different activities. This study declared the effectiveness of OA-α-cLA complex as a promising entity with anticancer activity, and these formulated OA-camel protein complexes constitute an auspicious approach for cancer remedy, particularly for breast cancer.
Topics: Animals; Antineoplastic Agents; Apoptosis; Caco-2 Cells; Camelus; Cell Cycle Checkpoints; Cell Proliferation; Chlorocebus aethiops; Drug Compounding; Female; Hep G2 Cells; Humans; Lactalbumin; MCF-7 Cells; Male; Milk; Neoplasms; Oleic Acid; Protein Kinase Inhibitors; Protein-Tyrosine Kinases; Vero Cells
PubMed: 28949782
DOI: 10.1080/07357907.2017.1373783 -
Journal of Agricultural and Food... Oct 2018The glucose glycation of α-lactalbumin and β-lactoglobulin at 50 °C in a glycerol-based liquid system was investigated to evaluate the effect of water activity on...
The glucose glycation of α-lactalbumin and β-lactoglobulin at 50 °C in a glycerol-based liquid system was investigated to evaluate the effect of water activity on glycation and site-specificity in a glycerol matrix. Glycation extent during the reaction was determined using the o-phthalaldehyde (OPA) method as well as ultraperformance liquid chromatography combined with electro-spray ionization mass spectrum (UPLC-ESI-MS) analysis. Glycation sites were identified by data-independent acquisition LC-MS (LC-MS). The surface potential achieved by PyMOL and the tertiary structure determined by circular dichroism (CD) were used to assist the analysis of the glycation site-specificity in the glycerol matrix. The water activity of glycerol solutions was negatively correlated to the glycerol concentration. Results showed that the initial glycation rate in glycerol matrix was fitted to a linear equation in the first 48 h. Glycation accelerated with the increase of glycerol concentration, namely, the decrease of water activity, regardless of the native structure of the protein. The glycation sites were identical at a similar DSP although achieved at different water activities, with 4 and 7 sites detected in α-lactalbumin and β-lactoglobulin, respectively. However, compared with the glycation sites in a water-based matrix, the site-specificity of glycation was affected by the glycerol matrix, depending on the native structure of the proteins. Glycation was prone to occur at the reactive sites distributed on the surface of the proteins, particularly in the region with positive potential.
Topics: Animals; Cattle; Chromatography, Liquid; Glucose; Glycerol; Glycosylation; Lactalbumin; Lactoglobulins; Spectrometry, Mass, Electrospray Ionization
PubMed: 30231611
DOI: 10.1021/acs.jafc.8b03544 -
Journal of the Science of Food and... Mar 2023The protein-polyphenol interaction mechanism has always been a research hotspot, but their interaction is affected by heat treatment, which is widely applied in food...
BACKGROUND
The protein-polyphenol interaction mechanism has always been a research hotspot, but their interaction is affected by heat treatment, which is widely applied in food processing. Moreover, the effects of microwave or water-bath heating on the protein-polyphenol interaction mechanism have been not clarified. The pasteurization condition (65 °C, 30 min) was selected to compare the effects of microwave or water bath on binding behavior, structure, and cell proliferation between α-lactalbumin (α-LA) and safflower yellow (SY), thus providing a guide for the selection of functional dairy processing conditions.
RESULTS
Microwave heat treatment of α-LA-SY resulted in stronger fluorescence quenching than that of conventional heat treatment. Moreover, the binding constant K of all α-LA-SY samples was augmented significantly after microwave or water bath treatment, and microwave-heated α-LA-SY showed the maximum K . Fourier transform infrared spectroscopy showed that microwave heating resulted in more ordered structures of α-LA into its disordered structures than water bath heating. However, the ferric reducing antioxidant power and chroma value of α-LA-SY were more reduced by microwave heating than by water bath heating. Moreover, microwave heating facilitated the cell proliferation of α-LA-SY compared with water bath treatment.
CONCLUSION
It was demonstrated that microwave heating promoted interaction between α-LA and SY more than water bath heating did. Microwave heat treatment was a safe and effective way to enhance the binding affinity of α-LA to SY, being a potential application in food industry. © 2022 Society of Chemical Industry.
Topics: Lactalbumin; Microwaves; Heating; Hot Temperature; Transcription Factors; Cell Proliferation; Water
PubMed: 36347624
DOI: 10.1002/jsfa.12325 -
Food & Function Feb 2022Bioactive peptides in bovine α-lactalbumin were isolated and identified, and the effects and mechanisms of peptide KILDK on insulin resistance in 3T3-L1 adipocytes were...
Bioactive peptides in bovine α-lactalbumin were isolated and identified, and the effects and mechanisms of peptide KILDK on insulin resistance in 3T3-L1 adipocytes were investigated. Mature 3T3-L1 adipocytes were stimulated with TNF-α to induce insulin resistance. Bovine α-lactalbumin hydrolysates (α-LAH) were subjected to stimulated gastrointestinal digestion and Caco-2 absorption, and GD-α-LAH and CA-α-LAH were obtained. Our results demonstrated that α-LAH, GD-α-LAH, and CA-α-LAH increased glucose uptake, enhanced Akt phosphorylation (Ser473), and decreased IRS-1 phosphorylation (Ser307) in insulin resistant 3T3-L1 adipocytes. Gel filtration chromatography and liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI MS/MS) were used to separate and identify bioactive peptides. The identified peptide KILDK attenuated insulin resistance in 3T3-L1 adipocytes, which was attributed to the suppression of JNK phosphorylation (Thr183/Tyr185). Moreover, KILDK downregulated pro-inflammatory genes through blocking NF-κB signaling. Our findings suggested that bovine α-LAH might be a potential ingredient against insulin resistance.
Topics: 3T3-L1 Cells; Animals; Caco-2 Cells; Cattle; Humans; Inflammation; Insulin Resistance; Lactalbumin; MAP Kinase Kinase 4; Mice; Peptides; Proto-Oncogene Proteins c-akt; Signal Transduction; Tumor Necrosis Factor-alpha
PubMed: 35142310
DOI: 10.1039/d1fo01217g