Phlorizin(PHL)is a natural compound with strong antioxidant properties mainly found in apples.In this paper,the interaction mechanism of PHL with pepsin and trypsin was comparatively evaluated by computer simulation,f...Phlorizin(PHL)is a natural compound with strong antioxidant properties mainly found in apples.In this paper,the interaction mechanism of PHL with pepsin and trypsin was comparatively evaluated by computer simulation,fluorescence spectra,circular dichroism(CD),and Fourier transform infrared(FT-IR)spectra at a molecular level.Fluorescence spectra showed that PHL quenches the pepsin/trypsin by static quenching.Thermodynamic parameters indicated that PHL binds to pepsin mainly through hydrogen bonds and van der Waals forces,and that of trypsin was electrostatic forces.The ground state complexes PHL and protease have a moderate affinity of 105 L/mol PHL binds more strongly to trypsin than to pepsin.CD and FT-IR spectra results showed that pepsin/trypsin decreased theβ-sheet content and slightly changed its secondary structure upon PHL.These experimental results are mutually verified with the predicted computer-aid simulation results.Upon PHL and trypsin binding,the antioxidant capacity of PHL was elevated.Nevertheless,the antioxidant capacity of PHL was decreased after binding to pepsin.This work elucidates the binding of PHL binding mechanisms to pepsin/trypsin and provides useful information for the digestion of PHL to improve the application of PHL in food processing.展开更多
During postmortem storage,fluoride in Antarctic krill can be enriched in the muscle.Trypsin,as the most important digestive enzyme in Antarctic krill with a high activity in low temperature,plays a potential role in t...During postmortem storage,fluoride in Antarctic krill can be enriched in the muscle.Trypsin,as the most important digestive enzyme in Antarctic krill with a high activity in low temperature,plays a potential role in this process.In this study,endogenous trypsin was purified and its properties were investigated.The involvement of trypsin in the generation of free fluoride from Antarctic krill cuticle was explored.Cuticle microstructure before and after hydrolysis was compared with scanning electron microscopy,and the ash samples of the hydrolyzed Antarctic krill cuticle were analyzed with X-ray diffraction,Fourier transform infrared spectroscopy,and electron dispersive spectroscopy,respectively.Mass spectrometry analysis and inhibition tests confirmed that the purified enzyme was endogenous trypsin.Results of the present study indicated that trypsin digestion caused the increases of the concentrations of both fluoride ions and free amino N simultaneously,while the protein coated on the cuticle surface was dissolved too.However,no compositional change was detected in the cuticle inorganic salts.These findings suggest that trypsin triggered free fluoride release from Antarctic krill cuticle.In addition,the kinetics of free fluoride release could be described by the equation C_(W)=(1-0.97^(-0.006t)-0.03e^(0.0558t))×337.53+10.50.The present study improved the understanding of the role of trypsin in free fluoride release from Antarctic krill cuticle,facilitating future studies aimed at reducing the fluoride content in krill protein during Antarctic krill processing.展开更多
AIM:To compare the safety and clinical outcomes of subconjunctival trypsin and dexamethasone(DEX)injections in the treatment of anterior chamber fibrin exudates in eyes with globe rupture following primary wound repai...AIM:To compare the safety and clinical outcomes of subconjunctival trypsin and dexamethasone(DEX)injections in the treatment of anterior chamber fibrin exudates in eyes with globe rupture following primary wound repair and vitrectomy.METHODS:A retrospective analysis included 42 males and 10 females(mean age 46.0±6.0y,range 34 to 58y)who underwent primary wound sutures and vitrectomy for globe rupture.Patients with pupil-covered fibrinous exudate or/and membrane in the anterior chamber were treated.On the first postoperative day,subconjunctival injections of either 5000 units(0.4 mL)of trypsin solution(n=25)or 0.5 mL(1 mg)DEX(n=27)were administered to accelerate exudate absorption.Efficacy was assessed by observing break time and partial absorption of the fibrin exudate membrane.Safety and comfort were evaluated by monitoring intraocular pressure(IOP),allergy,pain,and foreign body sensation.RESULTS:Both groups achieved 1/3 absorption of the anterior chamber fibrin exudate membrane,but the trypsin group exhibited shorter break time and partial absorption time compared to the DEX group(P<0.05).Trypsin treatment was also less irritating to patients.No adverse reactions were reported,and IOP remained stable.Visual acuity improved in both groups without statistical difference.CONCLUSION:Compared to DEX,trypsin demonstrates a shorter absorption time for the fibrin exudate membrane with a more comfortable process in treating pupil-covered fibrinous exudate or/and membrane after vitrectomy for globe rupture.展开更多
Herein,a novel method for fl uorometric detection of soybean trypsin inhibitor(SBTI)activity based on a water-soluble poly(diphenylacetylene)derivative was reported.Fluorescence quenching of the polymer via p-nitroani...Herein,a novel method for fl uorometric detection of soybean trypsin inhibitor(SBTI)activity based on a water-soluble poly(diphenylacetylene)derivative was reported.Fluorescence quenching of the polymer via p-nitroaniline,produced from the trypsin-catalyzed decomposition of N-benzoyl-DL-arginine-4-nitroanilide hydrochloride(L-BAPA),was well described using the Stern-Volmer equation.SBTI activity was quantitatively assessed based on changes in the fl uorescence intensity of the polymer.This strategy has several advantages,such as high sensitivity and ease of operation.Moreover,its applicability to other biochemical analyses is promising.展开更多
Trypsin,a novel superoxide scavenger,significantly enhances the storage quality of Hylocereus undatus(H.undatus).To elucidate the preservation mechanism of trypsin on H.undatus,a widely targeted metabolomic analysis,a...Trypsin,a novel superoxide scavenger,significantly enhances the storage quality of Hylocereus undatus(H.undatus).To elucidate the preservation mechanism of trypsin on H.undatus,a widely targeted metabolomic analysis,and transcriptomics analysis were conducted.Firstly,a total of 453 metabolites were identified,with organic acids and their derivatives constituting the largest proportion(25%).Amino acids and their metabolites,prominent among organic acids,were further analyzed.Among them,73 metabolites were associated with amino acids,and 37 exhibited significant differences.The most enriched Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway was arginine biosynthesis(map00220),with polyamine metabolites showing the most pronounced differences,particularly spermine(FC=1.7594).Compared with the control group,4-hydroxy-2-oxoglutaric acid was significantly upregulated(FC=2.117)in the process of spermine biosynthesis.Furthermore,the results of Gene Ontology(GO)and KEGG enrichment analysis of the H.undatus transcriptome profile revealed that trypsin treatment led to 187 differentially expressed genes associated with arginine.Both GO and KEGG analyses exhibited significant enrichment in the spermine biosynthetic process(GO:0006597)(map:00220)within the arginine biosynthesis pathway.Moreover,most enzymes and metabolites within the spermine biosynthesis pathway in H.undatus were upregulated.The results of the PPI network highlighted that ADC,SPDS,and SAMDC,among others,were pivotal proteins involved in trypsin-regulated arginine metabolism and spermine synthesis.This study revealed that trypsin could significantly delay postharvest senescence of H.undatus at room temperature.This effect might be attributed to trypsin triggering the synthesis of 4-hydroxy-2-oxoglutaric acid in the fruit peel,thereby promoting the biosynthesis of spermine and other polyamines.展开更多
By 30% - 60% (NH4)(2)SO4 fractional precipitation, anion-exchange chromatography on DEAE-Sepharose CL-6B, gel filtration on Sephacryl S-200 and anion-exchange chromatography on Waters AP-1 column (Protein(PM)-Pak DEAE...By 30% - 60% (NH4)(2)SO4 fractional precipitation, anion-exchange chromatography on DEAE-Sepharose CL-6B, gel filtration on Sephacryl S-200 and anion-exchange chromatography on Waters AP-1 column (Protein(PM)-Pak DEAE 15HR), a proteinase which can inactivate soybean trypsin inhibitor (STI) was purified from mung bean ( Vigna rabiata (L.) Wilezek) sprouts. Its molecular weight was estimated to be 29.8 kD by SDS-PAGE, and its K-m and V-max for STI were 769.2 N-alpha -benzoyl-L-arginine ethyl ester BAEE/mL and 115.3 BAEE . mL(-1) . min(-1) respectively. This proteinase was stable at temperatures lower than 50 degreesC and pH 6.5 - 8.5, and 90.91% STI activity of defatted soybean powder was inactivated by this preparation, with proteolytic activity 5 000 BAEE/mL at 50 degreesC and pH 8.0 in 4 h.展开更多
基金supported by the National Natural Science Foundation of China(21808020)the Applied Basic Research Program of Science&Technology Department of Sichuan Province(2018JY0151)。
文摘Phlorizin(PHL)is a natural compound with strong antioxidant properties mainly found in apples.In this paper,the interaction mechanism of PHL with pepsin and trypsin was comparatively evaluated by computer simulation,fluorescence spectra,circular dichroism(CD),and Fourier transform infrared(FT-IR)spectra at a molecular level.Fluorescence spectra showed that PHL quenches the pepsin/trypsin by static quenching.Thermodynamic parameters indicated that PHL binds to pepsin mainly through hydrogen bonds and van der Waals forces,and that of trypsin was electrostatic forces.The ground state complexes PHL and protease have a moderate affinity of 105 L/mol PHL binds more strongly to trypsin than to pepsin.CD and FT-IR spectra results showed that pepsin/trypsin decreased theβ-sheet content and slightly changed its secondary structure upon PHL.These experimental results are mutually verified with the predicted computer-aid simulation results.Upon PHL and trypsin binding,the antioxidant capacity of PHL was elevated.Nevertheless,the antioxidant capacity of PHL was decreased after binding to pepsin.This work elucidates the binding of PHL binding mechanisms to pepsin/trypsin and provides useful information for the digestion of PHL to improve the application of PHL in food processing.
基金supported by the Guangdong Provincial Key Laboratory of Aquatic Product Processing and Safety(No.GDPKLAPPS2005)the School Level Talent Project of Lingnan Normal University(No.ZL2021009)+2 种基金the Science and Technology Planning Project of Zhanjiang(No.2020A01040)the Study on the Preparation of Active Polypeptide from the Processing Waste of White Shrimp and its Fatigue Resistance(No.2021E05022)the Scientific Research Capacity Improvement Project of the Key Construction Discipline of Guangdong Province(No.2022ZD JS079).
文摘During postmortem storage,fluoride in Antarctic krill can be enriched in the muscle.Trypsin,as the most important digestive enzyme in Antarctic krill with a high activity in low temperature,plays a potential role in this process.In this study,endogenous trypsin was purified and its properties were investigated.The involvement of trypsin in the generation of free fluoride from Antarctic krill cuticle was explored.Cuticle microstructure before and after hydrolysis was compared with scanning electron microscopy,and the ash samples of the hydrolyzed Antarctic krill cuticle were analyzed with X-ray diffraction,Fourier transform infrared spectroscopy,and electron dispersive spectroscopy,respectively.Mass spectrometry analysis and inhibition tests confirmed that the purified enzyme was endogenous trypsin.Results of the present study indicated that trypsin digestion caused the increases of the concentrations of both fluoride ions and free amino N simultaneously,while the protein coated on the cuticle surface was dissolved too.However,no compositional change was detected in the cuticle inorganic salts.These findings suggest that trypsin triggered free fluoride release from Antarctic krill cuticle.In addition,the kinetics of free fluoride release could be described by the equation C_(W)=(1-0.97^(-0.006t)-0.03e^(0.0558t))×337.53+10.50.The present study improved the understanding of the role of trypsin in free fluoride release from Antarctic krill cuticle,facilitating future studies aimed at reducing the fluoride content in krill protein during Antarctic krill processing.
基金Supported by the Joint Construction Project of Henan Medical Science and Technology(No.LHGJ20220370)the Natural Science Foundation of Henan Province(No.232300420237).
文摘AIM:To compare the safety and clinical outcomes of subconjunctival trypsin and dexamethasone(DEX)injections in the treatment of anterior chamber fibrin exudates in eyes with globe rupture following primary wound repair and vitrectomy.METHODS:A retrospective analysis included 42 males and 10 females(mean age 46.0±6.0y,range 34 to 58y)who underwent primary wound sutures and vitrectomy for globe rupture.Patients with pupil-covered fibrinous exudate or/and membrane in the anterior chamber were treated.On the first postoperative day,subconjunctival injections of either 5000 units(0.4 mL)of trypsin solution(n=25)or 0.5 mL(1 mg)DEX(n=27)were administered to accelerate exudate absorption.Efficacy was assessed by observing break time and partial absorption of the fibrin exudate membrane.Safety and comfort were evaluated by monitoring intraocular pressure(IOP),allergy,pain,and foreign body sensation.RESULTS:Both groups achieved 1/3 absorption of the anterior chamber fibrin exudate membrane,but the trypsin group exhibited shorter break time and partial absorption time compared to the DEX group(P<0.05).Trypsin treatment was also less irritating to patients.No adverse reactions were reported,and IOP remained stable.Visual acuity improved in both groups without statistical difference.CONCLUSION:Compared to DEX,trypsin demonstrates a shorter absorption time for the fibrin exudate membrane with a more comfortable process in treating pupil-covered fibrinous exudate or/and membrane after vitrectomy for globe rupture.
基金The authors appreciate the support from the Zhe-jiang Province Lingyan Key R&D Project(No.2022C01177)the Zhejiang Administration for Market Regulation Eyas Program Cultiva-tion Project(No.CY2022355).
文摘Herein,a novel method for fl uorometric detection of soybean trypsin inhibitor(SBTI)activity based on a water-soluble poly(diphenylacetylene)derivative was reported.Fluorescence quenching of the polymer via p-nitroaniline,produced from the trypsin-catalyzed decomposition of N-benzoyl-DL-arginine-4-nitroanilide hydrochloride(L-BAPA),was well described using the Stern-Volmer equation.SBTI activity was quantitatively assessed based on changes in the fl uorescence intensity of the polymer.This strategy has several advantages,such as high sensitivity and ease of operation.Moreover,its applicability to other biochemical analyses is promising.
基金the National Key Research and Development Program of China(2017YFC1600802).
文摘Trypsin,a novel superoxide scavenger,significantly enhances the storage quality of Hylocereus undatus(H.undatus).To elucidate the preservation mechanism of trypsin on H.undatus,a widely targeted metabolomic analysis,and transcriptomics analysis were conducted.Firstly,a total of 453 metabolites were identified,with organic acids and their derivatives constituting the largest proportion(25%).Amino acids and their metabolites,prominent among organic acids,were further analyzed.Among them,73 metabolites were associated with amino acids,and 37 exhibited significant differences.The most enriched Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway was arginine biosynthesis(map00220),with polyamine metabolites showing the most pronounced differences,particularly spermine(FC=1.7594).Compared with the control group,4-hydroxy-2-oxoglutaric acid was significantly upregulated(FC=2.117)in the process of spermine biosynthesis.Furthermore,the results of Gene Ontology(GO)and KEGG enrichment analysis of the H.undatus transcriptome profile revealed that trypsin treatment led to 187 differentially expressed genes associated with arginine.Both GO and KEGG analyses exhibited significant enrichment in the spermine biosynthetic process(GO:0006597)(map:00220)within the arginine biosynthesis pathway.Moreover,most enzymes and metabolites within the spermine biosynthesis pathway in H.undatus were upregulated.The results of the PPI network highlighted that ADC,SPDS,and SAMDC,among others,were pivotal proteins involved in trypsin-regulated arginine metabolism and spermine synthesis.This study revealed that trypsin could significantly delay postharvest senescence of H.undatus at room temperature.This effect might be attributed to trypsin triggering the synthesis of 4-hydroxy-2-oxoglutaric acid in the fruit peel,thereby promoting the biosynthesis of spermine and other polyamines.
文摘By 30% - 60% (NH4)(2)SO4 fractional precipitation, anion-exchange chromatography on DEAE-Sepharose CL-6B, gel filtration on Sephacryl S-200 and anion-exchange chromatography on Waters AP-1 column (Protein(PM)-Pak DEAE 15HR), a proteinase which can inactivate soybean trypsin inhibitor (STI) was purified from mung bean ( Vigna rabiata (L.) Wilezek) sprouts. Its molecular weight was estimated to be 29.8 kD by SDS-PAGE, and its K-m and V-max for STI were 769.2 N-alpha -benzoyl-L-arginine ethyl ester BAEE/mL and 115.3 BAEE . mL(-1) . min(-1) respectively. This proteinase was stable at temperatures lower than 50 degreesC and pH 6.5 - 8.5, and 90.91% STI activity of defatted soybean powder was inactivated by this preparation, with proteolytic activity 5 000 BAEE/mL at 50 degreesC and pH 8.0 in 4 h.
