This study was designed to evaluate the bioactivities such asβ-glucosidase activity,α-galactosidase activity,and the growth behavior of the Lactobacillus cultures in soy milk medium.Ten Lactobacillus cultures were c...This study was designed to evaluate the bioactivities such asβ-glucosidase activity,α-galactosidase activity,and the growth behavior of the Lactobacillus cultures in soy milk medium.Ten Lactobacillus cultures were considered in this study.L.fermentum(M2)and L.casei(NK9)were selected due to their betterα-galactosidase,β-glucosidase activity and growth behavior in soy milk medium during fermentation.Further,soy milk fermented with M2 showed higher proteolytic activity(0.67 OD)and ACE-inhibitory(48.44%)than NK9(proteolytic activity:0.48 OD and ACE-inhibitory activity:41.33%).Bioactive peptides produced during the fermentation of soy milk using the selected Lactobacillus cultures were also identified with potent ACE-inhibitory activity by MALDI-TOF spectrometry,and the identified ACE inhibitory peptide sequences from fermented soy milk were characterized using Biopep database.展开更多
In this study,four types of Kas¸ar cheese were produced using raw milk(C),starter culture(Streptococcus thermophilus and Lactobacillus delbrueckii ssp.bulgaricus)(S),starter culture+probiotic Lactobacillus acidop...In this study,four types of Kas¸ar cheese were produced using raw milk(C),starter culture(Streptococcus thermophilus and Lactobacillus delbrueckii ssp.bulgaricus)(S),starter culture+probiotic Lactobacillus acidophilus ATCC 4356(SLA),and probiotic L.acidophilus ATCC 4356(LA)were produced and ripened at 6±2℃ for 150 days.The physicochemical,microbiological,proteolysis,electrophoretic(Urea-PAGE),ACE-inhibitory,and antioxidant activity analyses of vacuum-packed Kas¸ar cheeses were performed on 5,30,60,90,120,and 150 days of ripening.During ripening,no significant differences(p>0.05)were detected in total solids,fat,and protein values of Kas¸ar cheeses.Titratable acidity,water-soluble nitrogen(WSN),ripening index(RI),and electrophoresis results of cheese samples showed an increase while pH values decreased.Also,the degradation products ofαs1-casein were higher than those ofβ-casein.The number of probiotic L.acidophilus ATCC 4356 after scalding decreased by an average of 1.57 log cfu/g.However,probiotic bacteria count increased during ripening and maintained their viability.The addition of L.acidophilus ATCC 4356 improved the bioactivity,electrophoresis,titratable acidity,and ripening index results of Kas¸ar cheese.ACE-inhibitory activity of Kas¸ar cheeses increased during ripening and had the highest value on the 150th day of ripening.Furthermore,antioxidant activity of Kas¸ar cheeses increased on the 60th day of ripening and then decreased.Consequently,the combination of probiotic bacteria and starter culture had a positive effect on the physicochemical,microbiological,and proteolytic properties of Kas¸ar cheese.展开更多
Edible coatings offer many advantages in cheese coating for prolonging the shelf life and improving quality properties.Mould spoilage that occurs during ripening is one of the most important issues in Kas¸ar chee...Edible coatings offer many advantages in cheese coating for prolonging the shelf life and improving quality properties.Mould spoilage that occurs during ripening is one of the most important issues in Kas¸ar cheese.The current study focused on the effects of an edible coating solution based on quince seed mucilage(QSM)containing thyme oil(TO)in different ratios,on the surface moulding of Kas¸ar cheese and evaluated the physicochemical,microbiological,textural,and proteolysis characteristics of Kas¸ar cheese throughout the 120 days of ripening.Also,the effect of edible coating on the in-vitro Angiotensin-converting enzyme(ACE)-inhibitory activities of pH 4.6-soluble extracts of Kas¸ar cheeses was investigated.Results showed that the coating of Kas¸ar cheese with QSM and the increased levels of thyme oil significantly suppress mould growth throughout the ripening.Until 120^(th) days of ripening,counts of yeasts and moulds were lower<2 log cfu g^(-1)in coated cheese sample with QSM containing 2%TO.Coating with QSM caused an increase in moisture loss compared to vacuum-packed cheeses,however,didn’t change the normal ripening conditions such as microbiological,chemical,textural,and proteolysis properties of Kas¸ar cheese.As the ripening progressed,ACE-inhibitory activity increased in all samples,with the highest inhibition in vacuum-packed and coated cheeses.Overall,it was determined that the QSM coating showed good potential as a coating material for the protection of cheese shelf life and quality as well as for enhancing ACE-inhibitory activity.展开更多
In this study,Ulva prolifera protein was used for preparing angiotensin-I converting enzyme(ACE)-inhibitory peptide via virtual gastrointestinal digestion and in silico screening.Some parameters of the obtained peptid...In this study,Ulva prolifera protein was used for preparing angiotensin-I converting enzyme(ACE)-inhibitory peptide via virtual gastrointestinal digestion and in silico screening.Some parameters of the obtained peptide,such as inhibition kinetics,docking mechanism,stability,transport pathway,were explored by Lineweaver-Burk plots,molecular docking,in vitro stimulate gastrointestinal(GI)digestion and Caco-2 cells monolayer model,respectively.Then,a novel anti-ACE peptide LDF(IC_(50),(1.66±0.34)μmol/L)was screened and synthesized by chemical synthesis.It was a no-competitive inhibitor and its anti-ACE inhibitory effect mainly attributable to four Conventional Hydrogen Bonds and Zn701 interactions.It could keep activity during simulated GI digestion in vitro and was transported by peptide transporter PepT1 and passive-mediated mode.Besides,it could activate Endothelial nitric oxide synthase(eNOS)activity to promote the production of NO and reduce Endothelin-1(ET-1)secretion induced by AngiotensinⅡ(AngⅡ)in Human Umbilical Vein Endothelial Cells(HUVECs).Meanwhile,it could promote mice splenocytes proliferation in a concentration-dependent manner.Our study indicated that this peptide was a potential ingredient functioning on vasodilation and enhancing immunity.展开更多
Fish roes,rich in sialoglycoproteins,are often discarded,raising environmental concerns.The current study aimed to evaluate the antihypertensive effects of sialoglycoprotein hydrolysate produced from Labeo rohita roes...Fish roes,rich in sialoglycoproteins,are often discarded,raising environmental concerns.The current study aimed to evaluate the antihypertensive effects of sialoglycoprotein hydrolysate produced from Labeo rohita roes using Wistar rats.SGPH exhibited moderate but significant ACE-inhibitory activity(74.70±0.13%),DPPH radical scavenging activity(37.85±1.08%),ABTS radical scavenging activity(47.15±1.60%),and FRAP activity(27.13±0.03μM Trolox equivalents/g).The ACE-inhibition kinetics and gastrointestinal(GI)digestibility of SGPH demonstrated that SGPH was a competitive inhibitor and relatively stable in simulated GI conditions.SGPH revealed a moderate but significant antihypertensive effect on hypertensive(2K1C)Wistar rats in a dose-dependent manner by reducing systolic and diastolic blood pressure(SBP and DBP)and kidney ACE activity(P<0.05)after 4 weeks of treatment.Reduction of SBP,DBP,and kidney ACE activity ranged from 140±2-128±2 mmHg,100±3-82±3 mmHg,and 0.94±0.01-0.61±0.02μM HA/min/mg,respectively.Notably,SGPH elevated the superoxide dismutase and catalase activities in 2K1C rats(P<0.05),irrespective of SGPH dose,which could augment additional cardio-protection by minimizing oxidative stress.SGPH-treated rat groups displayed urine,haematology and serum biochemistry values in the normal range,indicating no adverse effects of SGPH.Further,histopathological evaluation elucidated that there was no incidence of organ abnormality in SGPH-treated rats,asserting that SGPH precluded vital organ injuries.Therefore,SGPH could be a promising antihypertensive ingredient that can be deployed in functional foods/nutraceuticals development for combating hypertension and its related diseases.展开更多
Defatted egg yolk is a by-product of phospholipid extraction from egg yolk.The hydrolysate of defatted egg yolk(EYH)obtained in this study showed multi-functions such as osteogenic,metal-chelating,antioxidant and ACEi...Defatted egg yolk is a by-product of phospholipid extraction from egg yolk.The hydrolysate of defatted egg yolk(EYH)obtained in this study showed multi-functions such as osteogenic,metal-chelating,antioxidant and ACEinhibitory activities.This study aimed to provide a scalable method for simultaneously enriching and separating different bioactive peptides from EYH using macroporous resins.Static adsorption and desorption assays indi-cated that DA201-C as the optimal resin.The loading sample pH value adjusted the enrichment effects,and then different bioactive peptides were separated in different ethanol gradient elution.Loading sample pH 7.5 showed the best effects with a total recovery of 87.65%(w/w,based on EYH).W-1 fraction enriched osteogenic peptides with promoted MC3T3-E1 proliferation and differentiation,and metal-chelating activities,Ethl-20%fraction enriched antioxidant peptides(TEAC=675.63μmol TE/g),Ethl-40%and Ethl-80%fractions enriched ACEinhibitory peptides(IC_(50)=0.26 mg/mL).More than twenty phosphopeptides were identified in W-1 by LC-MS/MS,which contributed to osteogenic activity.This research offered a simple and effective technique for large-scale enriching and separating osteogenic,metal-chelating,antioxidant,and ACE-inhibitory peptides from EYH,which is beneficial for the full utilization of defatted egg yolk powder.展开更多
Whey protein concentrate-80(WPC-80)fermented with L.fermentum(KGL4)(37℃)and S.cerevisiae(WBS2A)(25℃)was tested for ACE-inhibitory and antioxidant activities over different periods(12,24,36 and 48 h).Proteolytic acti...Whey protein concentrate-80(WPC-80)fermented with L.fermentum(KGL4)(37℃)and S.cerevisiae(WBS2A)(25℃)was tested for ACE-inhibitory and antioxidant activities over different periods(12,24,36 and 48 h).Proteolytic activity(OPA method)was used to optimize the growth conditions(inoculation rate,i.e.at 1.5%,2.0%,and 2.5%and incubation time,i.e.12,24,36,and 48 h)for peptide production.Results indicated that the highest amount of peptides was obtained at 7.24 mg/mL for KGL4(37℃,48 h)and 8.59 mg/mL for WBS2A(25℃,48 h).The whey protein fermentate inhibited the LPS-induced NO production,while enhanced production concentrations of TNF-α,IL-6,and IL-1β.Subsequently,SDS-PAGE,as well as Two-Dimensional(2D)gel electrophoresis methods,were applied for protein purification using water-soluble extracts(WSEs)of WPC-80 fermented by a combination of L.fermentum and S.cerevisiae.On SDS-PAGE,protein bands were observed in the range of 10-55 kDa,whereas on the 2D page,protein spots were in the range of 10-70 kDa.All the 2D spots were analyzed using RPLC/MS.WSEs of 3 kDa and 10 kDa permeates were used in RP-HPLC to identify distinct peptide fractions.The data from LC/MS was also characterized by utilizing ProteinPilot software.Further,different functional groups were also analyzed using FTIR investigation.The research aims to isolate and characterize novel ACE-inhibitory and antioxidative peptides from fermented WPC-80 produced by Lactobacillus fermentum and S.cerevisiae.展开更多
Chitosan and alginate have been used as promising biodegradable polymers in the oral delivery of bioactive compounds.In the present study,the stone fish-derived ACE-inhibitory peptide,ALGPQFY was encapsulated through ...Chitosan and alginate have been used as promising biodegradable polymers in the oral delivery of bioactive compounds.In the present study,the stone fish-derived ACE-inhibitory peptide,ALGPQFY was encapsulated through ionic gelation with alginate and polyelectrolyte complexation with sodium tripolyphosphate(TPP)cross-linked chitosan,to produce peptide loaded chitosan-coated alginate nanoparticles(PL-C-AL-NPs).The PL-C-AL-NPs were evaluated for physicochemical properties,morphology and thermal stability.The SBP-lowering effect due to the entrapped peptide was then studied on spontaneously hypertensive rats(SHRs).The results for physicochemical characterization revealed stable nanoparticles with small particle size and polydispersity index(pdi)of 212.60 nm and 0.327,and large values of entrapment efficiency(EE),peptide load(PL)and zeta potential of 74.48%,16.67%and+46.80 mV,respectively.Transmission electron microscopy(TEM)displayed homogenously-dispersed vesicular nanoparticles while differential scanning calorimetry indicated a high thermal stability of the nanoparticles with the melting temperature of 143.39℃.The PL-C-AL-NPs exhibited a dose dependent SBP-lowering effect that was significantly higher(p<0.05)compared to the free/unencapsulated peptide.This finding showed that,chitosan and alginate can be used in the design of nanocarriers to enhance the stability,bioavailability and antihypertensive efficacy of ALGPQFY which can be incorporated into functional foods and nutraceuticals for long-term hypertension management.展开更多
文摘This study was designed to evaluate the bioactivities such asβ-glucosidase activity,α-galactosidase activity,and the growth behavior of the Lactobacillus cultures in soy milk medium.Ten Lactobacillus cultures were considered in this study.L.fermentum(M2)and L.casei(NK9)were selected due to their betterα-galactosidase,β-glucosidase activity and growth behavior in soy milk medium during fermentation.Further,soy milk fermented with M2 showed higher proteolytic activity(0.67 OD)and ACE-inhibitory(48.44%)than NK9(proteolytic activity:0.48 OD and ACE-inhibitory activity:41.33%).Bioactive peptides produced during the fermentation of soy milk using the selected Lactobacillus cultures were also identified with potent ACE-inhibitory activity by MALDI-TOF spectrometry,and the identified ACE inhibitory peptide sequences from fermented soy milk were characterized using Biopep database.
基金supported by Atatürk University Coordination Unit of Scientific Research Projects within the scope of Graduate Thesis Projects.Project No:FDK-2021-8716(Project ID:8716).
文摘In this study,four types of Kas¸ar cheese were produced using raw milk(C),starter culture(Streptococcus thermophilus and Lactobacillus delbrueckii ssp.bulgaricus)(S),starter culture+probiotic Lactobacillus acidophilus ATCC 4356(SLA),and probiotic L.acidophilus ATCC 4356(LA)were produced and ripened at 6±2℃ for 150 days.The physicochemical,microbiological,proteolysis,electrophoretic(Urea-PAGE),ACE-inhibitory,and antioxidant activity analyses of vacuum-packed Kas¸ar cheeses were performed on 5,30,60,90,120,and 150 days of ripening.During ripening,no significant differences(p>0.05)were detected in total solids,fat,and protein values of Kas¸ar cheeses.Titratable acidity,water-soluble nitrogen(WSN),ripening index(RI),and electrophoresis results of cheese samples showed an increase while pH values decreased.Also,the degradation products ofαs1-casein were higher than those ofβ-casein.The number of probiotic L.acidophilus ATCC 4356 after scalding decreased by an average of 1.57 log cfu/g.However,probiotic bacteria count increased during ripening and maintained their viability.The addition of L.acidophilus ATCC 4356 improved the bioactivity,electrophoresis,titratable acidity,and ripening index results of Kas¸ar cheese.ACE-inhibitory activity of Kas¸ar cheeses increased during ripening and had the highest value on the 150th day of ripening.Furthermore,antioxidant activity of Kas¸ar cheeses increased on the 60th day of ripening and then decreased.Consequently,the combination of probiotic bacteria and starter culture had a positive effect on the physicochemical,microbiological,and proteolytic properties of Kas¸ar cheese.
文摘Edible coatings offer many advantages in cheese coating for prolonging the shelf life and improving quality properties.Mould spoilage that occurs during ripening is one of the most important issues in Kas¸ar cheese.The current study focused on the effects of an edible coating solution based on quince seed mucilage(QSM)containing thyme oil(TO)in different ratios,on the surface moulding of Kas¸ar cheese and evaluated the physicochemical,microbiological,textural,and proteolysis characteristics of Kas¸ar cheese throughout the 120 days of ripening.Also,the effect of edible coating on the in-vitro Angiotensin-converting enzyme(ACE)-inhibitory activities of pH 4.6-soluble extracts of Kas¸ar cheeses was investigated.Results showed that the coating of Kas¸ar cheese with QSM and the increased levels of thyme oil significantly suppress mould growth throughout the ripening.Until 120^(th) days of ripening,counts of yeasts and moulds were lower<2 log cfu g^(-1)in coated cheese sample with QSM containing 2%TO.Coating with QSM caused an increase in moisture loss compared to vacuum-packed cheeses,however,didn’t change the normal ripening conditions such as microbiological,chemical,textural,and proteolysis properties of Kas¸ar cheese.As the ripening progressed,ACE-inhibitory activity increased in all samples,with the highest inhibition in vacuum-packed and coated cheeses.Overall,it was determined that the QSM coating showed good potential as a coating material for the protection of cheese shelf life and quality as well as for enhancing ACE-inhibitory activity.
文摘In this study,Ulva prolifera protein was used for preparing angiotensin-I converting enzyme(ACE)-inhibitory peptide via virtual gastrointestinal digestion and in silico screening.Some parameters of the obtained peptide,such as inhibition kinetics,docking mechanism,stability,transport pathway,were explored by Lineweaver-Burk plots,molecular docking,in vitro stimulate gastrointestinal(GI)digestion and Caco-2 cells monolayer model,respectively.Then,a novel anti-ACE peptide LDF(IC_(50),(1.66±0.34)μmol/L)was screened and synthesized by chemical synthesis.It was a no-competitive inhibitor and its anti-ACE inhibitory effect mainly attributable to four Conventional Hydrogen Bonds and Zn701 interactions.It could keep activity during simulated GI digestion in vitro and was transported by peptide transporter PepT1 and passive-mediated mode.Besides,it could activate Endothelial nitric oxide synthase(eNOS)activity to promote the production of NO and reduce Endothelin-1(ET-1)secretion induced by AngiotensinⅡ(AngⅡ)in Human Umbilical Vein Endothelial Cells(HUVECs).Meanwhile,it could promote mice splenocytes proliferation in a concentration-dependent manner.Our study indicated that this peptide was a potential ingredient functioning on vasodilation and enhancing immunity.
基金Council of Scientific and Industrial Research(CSIR),New Delhi,India,for the research grant(CFTRI MLP-290)AV acknowledges the University Grants Commission,New Delhi,India,for the award of the CSIR-UGC fellowship.
文摘Fish roes,rich in sialoglycoproteins,are often discarded,raising environmental concerns.The current study aimed to evaluate the antihypertensive effects of sialoglycoprotein hydrolysate produced from Labeo rohita roes using Wistar rats.SGPH exhibited moderate but significant ACE-inhibitory activity(74.70±0.13%),DPPH radical scavenging activity(37.85±1.08%),ABTS radical scavenging activity(47.15±1.60%),and FRAP activity(27.13±0.03μM Trolox equivalents/g).The ACE-inhibition kinetics and gastrointestinal(GI)digestibility of SGPH demonstrated that SGPH was a competitive inhibitor and relatively stable in simulated GI conditions.SGPH revealed a moderate but significant antihypertensive effect on hypertensive(2K1C)Wistar rats in a dose-dependent manner by reducing systolic and diastolic blood pressure(SBP and DBP)and kidney ACE activity(P<0.05)after 4 weeks of treatment.Reduction of SBP,DBP,and kidney ACE activity ranged from 140±2-128±2 mmHg,100±3-82±3 mmHg,and 0.94±0.01-0.61±0.02μM HA/min/mg,respectively.Notably,SGPH elevated the superoxide dismutase and catalase activities in 2K1C rats(P<0.05),irrespective of SGPH dose,which could augment additional cardio-protection by minimizing oxidative stress.SGPH-treated rat groups displayed urine,haematology and serum biochemistry values in the normal range,indicating no adverse effects of SGPH.Further,histopathological evaluation elucidated that there was no incidence of organ abnormality in SGPH-treated rats,asserting that SGPH precluded vital organ injuries.Therefore,SGPH could be a promising antihypertensive ingredient that can be deployed in functional foods/nutraceuticals development for combating hypertension and its related diseases.
基金supported by the National Key Research and Development Program of China[2022YFD2101003].
文摘Defatted egg yolk is a by-product of phospholipid extraction from egg yolk.The hydrolysate of defatted egg yolk(EYH)obtained in this study showed multi-functions such as osteogenic,metal-chelating,antioxidant and ACEinhibitory activities.This study aimed to provide a scalable method for simultaneously enriching and separating different bioactive peptides from EYH using macroporous resins.Static adsorption and desorption assays indi-cated that DA201-C as the optimal resin.The loading sample pH value adjusted the enrichment effects,and then different bioactive peptides were separated in different ethanol gradient elution.Loading sample pH 7.5 showed the best effects with a total recovery of 87.65%(w/w,based on EYH).W-1 fraction enriched osteogenic peptides with promoted MC3T3-E1 proliferation and differentiation,and metal-chelating activities,Ethl-20%fraction enriched antioxidant peptides(TEAC=675.63μmol TE/g),Ethl-40%and Ethl-80%fractions enriched ACEinhibitory peptides(IC_(50)=0.26 mg/mL).More than twenty phosphopeptides were identified in W-1 by LC-MS/MS,which contributed to osteogenic activity.This research offered a simple and effective technique for large-scale enriching and separating osteogenic,metal-chelating,antioxidant,and ACE-inhibitory peptides from EYH,which is beneficial for the full utilization of defatted egg yolk powder.
文摘Whey protein concentrate-80(WPC-80)fermented with L.fermentum(KGL4)(37℃)and S.cerevisiae(WBS2A)(25℃)was tested for ACE-inhibitory and antioxidant activities over different periods(12,24,36 and 48 h).Proteolytic activity(OPA method)was used to optimize the growth conditions(inoculation rate,i.e.at 1.5%,2.0%,and 2.5%and incubation time,i.e.12,24,36,and 48 h)for peptide production.Results indicated that the highest amount of peptides was obtained at 7.24 mg/mL for KGL4(37℃,48 h)and 8.59 mg/mL for WBS2A(25℃,48 h).The whey protein fermentate inhibited the LPS-induced NO production,while enhanced production concentrations of TNF-α,IL-6,and IL-1β.Subsequently,SDS-PAGE,as well as Two-Dimensional(2D)gel electrophoresis methods,were applied for protein purification using water-soluble extracts(WSEs)of WPC-80 fermented by a combination of L.fermentum and S.cerevisiae.On SDS-PAGE,protein bands were observed in the range of 10-55 kDa,whereas on the 2D page,protein spots were in the range of 10-70 kDa.All the 2D spots were analyzed using RPLC/MS.WSEs of 3 kDa and 10 kDa permeates were used in RP-HPLC to identify distinct peptide fractions.The data from LC/MS was also characterized by utilizing ProteinPilot software.Further,different functional groups were also analyzed using FTIR investigation.The research aims to isolate and characterize novel ACE-inhibitory and antioxidative peptides from fermented WPC-80 produced by Lactobacillus fermentum and S.cerevisiae.
基金support being granted by the Malaysian Ministry of Science,Technology and Innovation(MOSTI)under the project No.02-01-04-SF 2309.
文摘Chitosan and alginate have been used as promising biodegradable polymers in the oral delivery of bioactive compounds.In the present study,the stone fish-derived ACE-inhibitory peptide,ALGPQFY was encapsulated through ionic gelation with alginate and polyelectrolyte complexation with sodium tripolyphosphate(TPP)cross-linked chitosan,to produce peptide loaded chitosan-coated alginate nanoparticles(PL-C-AL-NPs).The PL-C-AL-NPs were evaluated for physicochemical properties,morphology and thermal stability.The SBP-lowering effect due to the entrapped peptide was then studied on spontaneously hypertensive rats(SHRs).The results for physicochemical characterization revealed stable nanoparticles with small particle size and polydispersity index(pdi)of 212.60 nm and 0.327,and large values of entrapment efficiency(EE),peptide load(PL)and zeta potential of 74.48%,16.67%and+46.80 mV,respectively.Transmission electron microscopy(TEM)displayed homogenously-dispersed vesicular nanoparticles while differential scanning calorimetry indicated a high thermal stability of the nanoparticles with the melting temperature of 143.39℃.The PL-C-AL-NPs exhibited a dose dependent SBP-lowering effect that was significantly higher(p<0.05)compared to the free/unencapsulated peptide.This finding showed that,chitosan and alginate can be used in the design of nanocarriers to enhance the stability,bioavailability and antihypertensive efficacy of ALGPQFY which can be incorporated into functional foods and nutraceuticals for long-term hypertension management.
