Bovineα-lactalbumin(BLA)induced severe cow's milk allergy.In this study,a novel strategy combining ultrasonication,performed before glycation,and phosphorylation was proposed to reduce BLA allergenicity.Result sh...Bovineα-lactalbumin(BLA)induced severe cow's milk allergy.In this study,a novel strategy combining ultrasonication,performed before glycation,and phosphorylation was proposed to reduce BLA allergenicity.Result showed that IgE-and IgG-binding capacities and the release rates of histamine and interleukin-6 from RBL-2 H3 were reduced.Moreover,intrinsic fluorescence intensity and surface hydrophobicity were decreased,whereas glycated sites(R10,N44,K79,K108,N102 and K114)and phosphorylated sites(Y36 and S112)of BLA were increased.Minimum allergenicity was detected during BLA treatment after ultrasonic prior to glycation and subsequent phosphorylation because of considerable increase in glycated and phosphorylated sites.Therefore,the decrease in allergenicity of BLA,the effect correlated well with the shielding effect of glycated sites combined with phosphorylated sites and the conformational changes.This study provides important theoretical foundations for improving and using the ultrasonic technology combined with protein modification in allergenic protein processing.展开更多
Reversed-phase high-performance liquid chromatography （RP-HPLC） method with C4 column and C18 column for analyzing β-lactoglobulin and α-lactalbumin in bovine milk was developed and the performance and characteris...Reversed-phase high-performance liquid chromatography （RP-HPLC） method with C4 column and C18 column for analyzing β-lactoglobulin and α-lactalbumin in bovine milk was developed and the performance and characteristic of two columns were compared. Shiseido Proteonavi C4 column （250 mm×4.6 mm×5μm） and Shiseido CAPCELL PAK SG 300 C18 column （250 mm× 4.6 mm×5 μm） were used in the experiment. Phase A was composed of 0.1% （V/V） trifluoroacetic acid （TFA） in ultrapure water and Phase B （organic phase） was composed of 0.1% TFA in acetonitrile. Gradient elution was taken. Flow rate was 1 mL min-1. The detection wavelength was 215 nm. The injection volume was 20 μL and the column temperature was 30℃. The results showed that linear relationship was good and recovery of α-lactalbumin and β-lactoglobulin was 86.12%-104.38%, C18 column had stronger ability to resist acid and more stable, and the method with C4 column had excellent sensitivities and good separation.展开更多
α-lactalbumin (α-LA) might increase its antioxidant potential after hydrolysis. In particular, low molecular weight (LMW) peptides showed greater antioxidant capacity. Different hydrolysis conditions with Alcalase e...α-lactalbumin (α-LA) might increase its antioxidant potential after hydrolysis. In particular, low molecular weight (LMW) peptides showed greater antioxidant capacity. Different hydrolysis conditions with Alcalase enzyme were optimized with a composite central design and surface methodology. Sample obtained after 0.1% (w/w enzyme:substrate), 60 min hydrolysis, ultrafiltrated with membranes of 3 kDa (named 4 LMW), showed the greatest antioxidant values: 1.574 ± 0.060 and 1.636 ± 0.076 μmolTE/mg of protein for ABTS and ORAC-FL, respectively. Sample 4 LMW produced mild ACE inhibition capacity, 22% related to Captopril. 4 LMW was submitted to in vitro gastrointestinal conditions using α-amylase, pepsin, pancreatin and bile-extract;its antioxidant capacity was enhanced by the shorter peptides released, confirmed by SE-HPLC. Antioxidant capacity of digested 4 LMW sample (D 4 LMW) was 1.743 ± 0.086 and 2.542 ± 0.245 μmolTE/mg of protein for ABTS and ORAC-FL, respectively, showing improvement on bioaccessibility. Intestinal cells viability was higher for D 4 LMW.展开更多
Casein andα-lactalbumin(α-LA)are the main allergens in cow's milk,which can affect the skin,respiratory system,and gastrointestinal tract,even cause anaphylactic shock.Developing a rapid and sensitive detection ...Casein andα-lactalbumin(α-LA)are the main allergens in cow's milk,which can affect the skin,respiratory system,and gastrointestinal tract,even cause anaphylactic shock.Developing a rapid and sensitive detection method of casein andα-LA is still pursued.Herein,a surface-enhanced Raman scattering based lateral flow assay(SERS-LFA)method for rapid and highly sensitive detection of milk allergens in food was established for effectively preventing allergic symptoms.Gold@silver nanoparticles(Au@Ag NPs)were synthesized as SERS active substrate to prepare the antibody-modified SERS Probe and SERS-LFA strips toward casein andα-LA were assembled according to the sandwich mode.The detection results were calculated according to colorimetric and Raman signal.The introduction of SRES signal significantly increased the sensitivity of detection with the limit of detection(LOD)of 0.19 ng/mL and 1.74 pg/mL,and exhibited an excellent linear relationship within the range of 0.55-791.50 ng/mL and 0.1 pg/mL-100.0 ng/mL for casein andα-LA,respectively.Furthermore,SERS-LFA strips was highly specific with the recovery rates for corresponding80.36%-105.12%and 85.73%-118.22%for casein andα-LA,respectively.Therefore,the SERS-LFA could be in great potential to develop a unique allergen detection method.展开更多
Burn injury is a serious public health problem and scientists are continuously aiming to develop promising biomimetic dressings for effective burn wound management.In this study,a greater efficacy in burn wound healin...Burn injury is a serious public health problem and scientists are continuously aiming to develop promising biomimetic dressings for effective burn wound management.In this study,a greater efficacy in burn wound healing and the associated mechanisms ofα-lactalbumin(ALA)based electrospun nanofibrous scaffolds(ENs)as compared to other regenerative protein scaffolds were established.Bovine serum albumin(BSA),collagen type I(COL),lysozyme(LZM)and ALA were separately blended with poly(ε-caprolactone)(PCL)to fabricate four different composite ENs(LZM/PCL,BSA/PCL,COL/PCL and ALA/PCL ENs).The hydrophilic composite scaffolds exhibited an enhancedwettability and variablemechanical properties.The ALA/PCL ENs demonstrated higher levels of fibroblast proliferation and adhesion than the other composite ENs.As compared to PCL ENs and other composite scaffolds,the ALA/PCL ENs also promoted a better maturity of the regenerative skin tissues and showed a comparable wound healing effect to Collagen sponge^(■)on third-degree burn model.The enhanced wound healing activity of ALA/PCL ENs compared to other ENs could be attributed to their ability to promote serotonin production at wound sites.Collectively,this investigation demonstrated that ALA is a unique protein with a greater potential for burn wound healing as compared to other regenerative proteins when loaded in the nanofibrous scaffolds.展开更多
基金supported by Science Foundation for Young Scientists of Jiangxi Province(20202BABL215027)National Natural Science Foundation of China(31960457)。
文摘Bovineα-lactalbumin(BLA)induced severe cow's milk allergy.In this study,a novel strategy combining ultrasonication,performed before glycation,and phosphorylation was proposed to reduce BLA allergenicity.Result showed that IgE-and IgG-binding capacities and the release rates of histamine and interleukin-6 from RBL-2 H3 were reduced.Moreover,intrinsic fluorescence intensity and surface hydrophobicity were decreased,whereas glycated sites(R10,N44,K79,K108,N102 and K114)and phosphorylated sites(Y36 and S112)of BLA were increased.Minimum allergenicity was detected during BLA treatment after ultrasonic prior to glycation and subsequent phosphorylation because of considerable increase in glycated and phosphorylated sites.Therefore,the decrease in allergenicity of BLA,the effect correlated well with the shielding effect of glycated sites combined with phosphorylated sites and the conformational changes.This study provides important theoretical foundations for improving and using the ultrasonic technology combined with protein modification in allergenic protein processing.
基金Supported by the Project of Science & Technology Plans in Heilongjiang Province in the 11th Five-year Period (GB07B407)
文摘Reversed-phase high-performance liquid chromatography （RP-HPLC） method with C4 column and C18 column for analyzing β-lactoglobulin and α-lactalbumin in bovine milk was developed and the performance and characteristic of two columns were compared. Shiseido Proteonavi C4 column （250 mm×4.6 mm×5μm） and Shiseido CAPCELL PAK SG 300 C18 column （250 mm× 4.6 mm×5 μm） were used in the experiment. Phase A was composed of 0.1% （V/V） trifluoroacetic acid （TFA） in ultrapure water and Phase B （organic phase） was composed of 0.1% TFA in acetonitrile. Gradient elution was taken. Flow rate was 1 mL min-1. The detection wavelength was 215 nm. The injection volume was 20 μL and the column temperature was 30℃. The results showed that linear relationship was good and recovery of α-lactalbumin and β-lactoglobulin was 86.12%-104.38%, C18 column had stronger ability to resist acid and more stable, and the method with C4 column had excellent sensitivities and good separation.
文摘α-lactalbumin (α-LA) might increase its antioxidant potential after hydrolysis. In particular, low molecular weight (LMW) peptides showed greater antioxidant capacity. Different hydrolysis conditions with Alcalase enzyme were optimized with a composite central design and surface methodology. Sample obtained after 0.1% (w/w enzyme:substrate), 60 min hydrolysis, ultrafiltrated with membranes of 3 kDa (named 4 LMW), showed the greatest antioxidant values: 1.574 ± 0.060 and 1.636 ± 0.076 μmolTE/mg of protein for ABTS and ORAC-FL, respectively. Sample 4 LMW produced mild ACE inhibition capacity, 22% related to Captopril. 4 LMW was submitted to in vitro gastrointestinal conditions using α-amylase, pepsin, pancreatin and bile-extract;its antioxidant capacity was enhanced by the shorter peptides released, confirmed by SE-HPLC. Antioxidant capacity of digested 4 LMW sample (D 4 LMW) was 1.743 ± 0.086 and 2.542 ± 0.245 μmolTE/mg of protein for ABTS and ORAC-FL, respectively, showing improvement on bioaccessibility. Intestinal cells viability was higher for D 4 LMW.
基金supported by the Science and Technology Innovation Action Plan of Shanghai,China(17391901302)the National Key R&D Program of China(2018YFC1604401)。
文摘Casein andα-lactalbumin(α-LA)are the main allergens in cow's milk,which can affect the skin,respiratory system,and gastrointestinal tract,even cause anaphylactic shock.Developing a rapid and sensitive detection method of casein andα-LA is still pursued.Herein,a surface-enhanced Raman scattering based lateral flow assay(SERS-LFA)method for rapid and highly sensitive detection of milk allergens in food was established for effectively preventing allergic symptoms.Gold@silver nanoparticles(Au@Ag NPs)were synthesized as SERS active substrate to prepare the antibody-modified SERS Probe and SERS-LFA strips toward casein andα-LA were assembled according to the sandwich mode.The detection results were calculated according to colorimetric and Raman signal.The introduction of SRES signal significantly increased the sensitivity of detection with the limit of detection(LOD)of 0.19 ng/mL and 1.74 pg/mL,and exhibited an excellent linear relationship within the range of 0.55-791.50 ng/mL and 0.1 pg/mL-100.0 ng/mL for casein andα-LA,respectively.Furthermore,SERS-LFA strips was highly specific with the recovery rates for corresponding80.36%-105.12%and 85.73%-118.22%for casein andα-LA,respectively.Therefore,the SERS-LFA could be in great potential to develop a unique allergen detection method.
基金supported by the Liaoning Pan Deng Xue Zhe Scholar(No.XLYC2002061)the Overseas Expertise Introduction Project for Discipline Innovation(“111 Project”)(No.D20029)+5 种基金X.G.acknowledges the Educational Department of Liaoning Province(grant No.LJKZ0925),(Youth Project,grant No.LJKQZ2021035)the international postdoctoral exchange fellowship program(grant No.PC2021047)H.B.thanks the financial support from National Natural Science Foundation of China(grant No.82050410448)Fellowship of China Postdoctoral Science Foundation(grant No.2021MD703857)D.Cun acknowledges financial support from Ministry of Education Chunhui Program(2020)VF acknowledges VILLUM FONDEN for supporting the project via the Villum Young Investigator Grant(grant No.19175).
文摘Burn injury is a serious public health problem and scientists are continuously aiming to develop promising biomimetic dressings for effective burn wound management.In this study,a greater efficacy in burn wound healing and the associated mechanisms ofα-lactalbumin(ALA)based electrospun nanofibrous scaffolds(ENs)as compared to other regenerative protein scaffolds were established.Bovine serum albumin(BSA),collagen type I(COL),lysozyme(LZM)and ALA were separately blended with poly(ε-caprolactone)(PCL)to fabricate four different composite ENs(LZM/PCL,BSA/PCL,COL/PCL and ALA/PCL ENs).The hydrophilic composite scaffolds exhibited an enhancedwettability and variablemechanical properties.The ALA/PCL ENs demonstrated higher levels of fibroblast proliferation and adhesion than the other composite ENs.As compared to PCL ENs and other composite scaffolds,the ALA/PCL ENs also promoted a better maturity of the regenerative skin tissues and showed a comparable wound healing effect to Collagen sponge^(■)on third-degree burn model.The enhanced wound healing activity of ALA/PCL ENs compared to other ENs could be attributed to their ability to promote serotonin production at wound sites.Collectively,this investigation demonstrated that ALA is a unique protein with a greater potential for burn wound healing as compared to other regenerative proteins when loaded in the nanofibrous scaffolds.
