Proteases,with their extensive sources and remarkable characteristics such as high catalytic efficiency,substrate specificity,and species diversity,have long attracted widespread attention and are widely applied in va...Proteases,with their extensive sources and remarkable characteristics such as high catalytic efficiency,substrate specificity,and species diversity,have long attracted widespread attention and are widely applied in various fields including food processing,detergent production,pharmaceutical,and environmental protection;these enzymes can hydrolyze proteins into peptides and amino acids,thereby participating in crucial physiological activities like digestion and immune regulation,and playing an indispensable role in maintaining the health and daily life of organisms.Moreover,through artificial synthesis of the required proteases,it is possible to achieve efficient large-scale expression and production,which significantly reduces industrial costs,making them more economically viable in practical applications.This paper provides a comprehensive review of proteases,covering their classification,sources,structure-activity relationships,and industrial applications,and constructs a closed-loop analytical framework based on“basic characteristics,production technology,and practical application”to systematically organize and analyze the relevant information;in particular,it quantitatively compares the advantages and defects of expression systems using Escherichia coli,yeast,and Bacillus subtilis,which not only deepens the understanding of these systems but also provides valuable theoretical support for the rational selection of expression vectors in different scenarios,and introduces their development prospects in food,medicine,environmental protection,and related fields.The insights provided herein offer specific directions for future applied research on artificially engineered proteases.展开更多
Proteases are essential for homeostasis,and their primary function is proteolytic in extracellular and intracellular compartments.The deregulation of expression,abundance,and activity of proteases has been related to ...Proteases are essential for homeostasis,and their primary function is proteolytic in extracellular and intracellular compartments.The deregulation of expression,abundance,and activity of proteases has been related to several pathologies,including cancer.This deregulation contributes to their pro-tumorigenic activity since they participate in the degradation of extracellular matrix components and adhesion molecules,and the activation of growth factors.However,some proteases,such as ADAM metallopeptidase with thrombospondin type 1 motif 8 and kallikrein-related peptidases 5 and 10,have emerged as tumor suppressors due to their antitumoral actions in specific cancer contexts.In this article,we discuss the antitumoral effects of ADAM metallopeptidase with thrombospondin type 1 motif 8,kallikrein-related peptidases 5 and 10 that have been described to date,suggesting their potential use as novel biomarkers and therapeutic targets in cancer.展开更多
Snake venoms,especially those from the two subfamilies,Crotalinae and Viperinae,contained a lot of serine proteases. They were responsible for the hemorrhage,shock,or disorder of blood coagulation after envenomation. ...Snake venoms,especially those from the two subfamilies,Crotalinae and Viperinae,contained a lot of serine proteases. They were responsible for the hemorrhage,shock,or disorder of blood coagulation after envenomation. They acted,by activating,inactivating,or other converting effects,on almost all the components of hemostatic and fibrinolytic systems. Their sequences were homologous to trypsin-kallikrein serine proteases. Variation of primary sequences out of active center results in the difference of substrate specificities and the further difference of biological and pharmacological activities. Because of their common and unique properties compared to their physiological corresponding factors,snake venom proteases are proved to be an excellent model for the study of protease substrate discriminating mechanism. Furthermore,they have found an important position both in basic research and application of hemostasis and thrombosis in clinic.展开更多
[Objective] The study aimed to investigate the activity and gene expression of caspase-like proteases in tobacco leaves growing under different light qualities. [Method] By covering tobacco plants with white, red, yel...[Objective] The study aimed to investigate the activity and gene expression of caspase-like proteases in tobacco leaves growing under different light qualities. [Method] By covering tobacco plants with white, red, yellow, blue and purple films to obtain different light quality, the changes of chlorophyll content, activity and gene expression of caspase-like proteases in the tobacco leaves were studied. [Results] Compared with treatments of white, red and yellow film, blue and purple films delayed the decrease of chlorophyll content and senescence of tobacco leaves at the late growth stage, and relatively lowered the activity and gene expression of caspase-like proteases during growth, development and senescence periods. [Conclusion] Different light qualities exhibited various effects on the growth, development and senescence of tobacco leaves, possibly by affecting the activity and gene expression of caspase-like proteases to some extent.展开更多
Celiac disease(CD)is a globally prevalent autoimmune disorder,affecting 1.4%of the population with 7.5%annual incidence increase.In genetically predisposed individuals,gliadin antigen can trigger an immune response in...Celiac disease(CD)is a globally prevalent autoimmune disorder,affecting 1.4%of the population with 7.5%annual incidence increase.In genetically predisposed individuals,gliadin antigen can trigger an immune response in the small intestine,causing villous atrophy that leads to severe symptoms.Current treatment of CD relies on strict gluten-free diets.Neprosin from Nepenthes species is one of the most promising enzymes for the detoxification of gliadin antigen.However,the potential of neprosin in gluten-safe food production has yet to be explored.In this study,neprosin from Nepenthes rafflesiana(NrNpr1)was recombinantly expressed with chicken lysozyme signal peptide in the baker's yeast Saccharomyces cerevisiae expression host.The culture media of the bioengineered S.cerevisiae can degrade pure gliadin substrate and gliadin in wheat flour.The live yeast culture was demonstrated to degrade gliadin in wheat flour dough.The purified recombinant NrNpr1(rNrNpr1)enzyme was successfully characterized to have an optimal pH of 2.5,optimal temperature of 50℃,thermal tolerance of 0-65℃,and can digest all immunogenic epitopes inα-gliadin 33-mer when tested in vitro.ELISA test proved that rNrNpr1 can detoxify 750 mg/L gliadin to a gluten-free level(<20 mg/L)after 6 h of incubation.This study characterized neprosin from N.rafflesiana and report the bioengineering of S.cerevisiae with recombinant neprosin.We demonstrated that rNrNpr1 has promising potential applications in revolutionary gluten-safe food production,which needs further optimization and formulation in bakery.展开更多
Research on marine bioactive peptides has mainly focused on characterization of peptides in hydrolysates prepared with commercial industrial enzymes and the usefulness of such hydrolysates in health and functional foo...Research on marine bioactive peptides has mainly focused on characterization of peptides in hydrolysates prepared with commercial industrial enzymes and the usefulness of such hydrolysates in health and functional foods. However, a relevant question is whether digestion of fish proteins with gastrointestinal proteases per se generates peptides that also can have health promoting properties and can reduce, e.g., diabetes 2, inflammation and hypertension either in relation to gastrointestinal digestion or as alternative to industrial proteases. The aim of the study was to investigate hydrolysates obtained from in vitro sequential digestion of salmon muscle and skin with gastrointestinal proteases including pepsin, pancreatic and pancreatic ＋ mucosal proteases for their ability to scavenge ABTS^＋ radicals and inhibit activity of angiotensin I-converting enzyme （ACE） and dipeptidyl peptidase 4 （DPP-4）. Furthermore, it was the aim to study the inhibitory mechanism and stability towards ACE and DPP-4 activity. Analysis of〈 10 kDa hydrolysates showed that gastrointestinal proteases generated peptides with clear radical scavenging activity and DPP-4 and ACE inhibiting activity as well. Hydrolysates from pepsin digestion exhibited the lowest ECso values for radical scavenging activity and ACE inhibition, whereas ECso increased in hydrolysates after subsequent digestion with pancreatic and mucosal proteases. Interestingly, ECso values for the DPP-4 inhibition were hardly affected by sequential digestion. Inhibition modes for the muscle hydrolysates were both competitive and non-competitive, but prolonged incubation showed that the inhibitory properties were unstable and therefore they were probably digested as competitive substrates by gastrointestinal proteases.展开更多
Objectives Dysregulated osteoclast function contributes to skeletal diseases.However,the specific ubiquitination regulators of the osteoclastogenesis repressor MafB,particularly at the post-translational level,remain ...Objectives Dysregulated osteoclast function contributes to skeletal diseases.However,the specific ubiquitination regulators of the osteoclastogenesis repressor MafB,particularly at the post-translational level,remain undefined.This study aims to identify ubiquitin-specific proteases(USPs)that deubiquitinate MafB and enhance its stability.Methods We constructed a MafB-conjugated luciferase and overexpressed 40 individual USPs,measuring changes in luciferase activity.The identified USP was overexpressed in human CD14^(+) peripheral blood mononuclear cells(PBMCs)to evaluate its effect.Osteoclast differentiation was assessed through osteoclast marker Integrin alpha-V(CD51)staining and Western blot analysis.Co-immunoprecipitation(co-IP)was performed to assess the interplay.The influence on MafB ubiquitination and degradation was evaluated via immunoprecipitation and Western blot.Finally,MafB was knocked down in the USP-overexpressing PBMCs to analyze its effect on osteoclast differentiation.Results Overexpression of ubiquitin-specific protease 29(USP29)significantly increased MafB expression by approximately 75%(p<0.0001).Elevated USP29 levels strongly inhibited osteoclastic differentiation in CD14^(+) PBMCs(p<0.0001).USP29 was found to interact with MafB,markedly reducing its ubiquitination and subsequent degradation in PBMCs(p<0.001).Knocking down MafB in USP29-overexpressing PBMCs alleviated the inhibitory effect of USP29 on osteoclastogenesis.Conclusion USP29 acts as a potent stabilizer of MafB,inhibiting osteoclastogenesis in human CD14^(+) PBMCs,at least in part,by enhancing MafB stability.These findings expand our understanding of USP29’s role and the post-translational regulation of MafB.Furthermore,USP29 serves as a vital factor that controls osteoclast differentiation,and its regulatory function is at least partially mediated by deubiquitinating and stabilizing MafB.展开更多
文摘Proteases,with their extensive sources and remarkable characteristics such as high catalytic efficiency,substrate specificity,and species diversity,have long attracted widespread attention and are widely applied in various fields including food processing,detergent production,pharmaceutical,and environmental protection;these enzymes can hydrolyze proteins into peptides and amino acids,thereby participating in crucial physiological activities like digestion and immune regulation,and playing an indispensable role in maintaining the health and daily life of organisms.Moreover,through artificial synthesis of the required proteases,it is possible to achieve efficient large-scale expression and production,which significantly reduces industrial costs,making them more economically viable in practical applications.This paper provides a comprehensive review of proteases,covering their classification,sources,structure-activity relationships,and industrial applications,and constructs a closed-loop analytical framework based on“basic characteristics,production technology,and practical application”to systematically organize and analyze the relevant information;in particular,it quantitatively compares the advantages and defects of expression systems using Escherichia coli,yeast,and Bacillus subtilis,which not only deepens the understanding of these systems but also provides valuable theoretical support for the rational selection of expression vectors in different scenarios,and introduces their development prospects in food,medicine,environmental protection,and related fields.The insights provided herein offer specific directions for future applied research on artificially engineered proteases.
基金Supported by Colegio de Ciencia y Tecnologia de la Universidad Autónoma de la Ciudad de México,No.UACM-CCyT-2025-CON-11.
文摘Proteases are essential for homeostasis,and their primary function is proteolytic in extracellular and intracellular compartments.The deregulation of expression,abundance,and activity of proteases has been related to several pathologies,including cancer.This deregulation contributes to their pro-tumorigenic activity since they participate in the degradation of extracellular matrix components and adhesion molecules,and the activation of growth factors.However,some proteases,such as ADAM metallopeptidase with thrombospondin type 1 motif 8 and kallikrein-related peptidases 5 and 10,have emerged as tumor suppressors due to their antitumoral actions in specific cancer contexts.In this article,we discuss the antitumoral effects of ADAM metallopeptidase with thrombospondin type 1 motif 8,kallikrein-related peptidases 5 and 10 that have been described to date,suggesting their potential use as novel biomarkers and therapeutic targets in cancer.
文摘Snake venoms,especially those from the two subfamilies,Crotalinae and Viperinae,contained a lot of serine proteases. They were responsible for the hemorrhage,shock,or disorder of blood coagulation after envenomation. They acted,by activating,inactivating,or other converting effects,on almost all the components of hemostatic and fibrinolytic systems. Their sequences were homologous to trypsin-kallikrein serine proteases. Variation of primary sequences out of active center results in the difference of substrate specificities and the further difference of biological and pharmacological activities. Because of their common and unique properties compared to their physiological corresponding factors,snake venom proteases are proved to be an excellent model for the study of protease substrate discriminating mechanism. Furthermore,they have found an important position both in basic research and application of hemostasis and thrombosis in clinic.
基金Supported by National Natural Science Foundation of China(30460016)Science and Technology Plan from Yunnan Branch Office of China National Tobacco Corporation(2011YN03,2010YN03,07A01)~~
文摘[Objective] The study aimed to investigate the activity and gene expression of caspase-like proteases in tobacco leaves growing under different light qualities. [Method] By covering tobacco plants with white, red, yellow, blue and purple films to obtain different light quality, the changes of chlorophyll content, activity and gene expression of caspase-like proteases in the tobacco leaves were studied. [Results] Compared with treatments of white, red and yellow film, blue and purple films delayed the decrease of chlorophyll content and senescence of tobacco leaves at the late growth stage, and relatively lowered the activity and gene expression of caspase-like proteases during growth, development and senescence periods. [Conclusion] Different light qualities exhibited various effects on the growth, development and senescence of tobacco leaves, possibly by affecting the activity and gene expression of caspase-like proteases to some extent.
基金supported by the Malaysian Ministry of Higher Education Fundamental Research Grant Scheme(FRGS/1/2024/STG01/UKM/02/5)Universiti Kebangsaan Malaysia Research University Grant(GUP-2024-092)awarded to Hoe-Han Gohsupported by Universiti Kebangsaan Malaysia postdoctoral research scheme。
文摘Celiac disease(CD)is a globally prevalent autoimmune disorder,affecting 1.4%of the population with 7.5%annual incidence increase.In genetically predisposed individuals,gliadin antigen can trigger an immune response in the small intestine,causing villous atrophy that leads to severe symptoms.Current treatment of CD relies on strict gluten-free diets.Neprosin from Nepenthes species is one of the most promising enzymes for the detoxification of gliadin antigen.However,the potential of neprosin in gluten-safe food production has yet to be explored.In this study,neprosin from Nepenthes rafflesiana(NrNpr1)was recombinantly expressed with chicken lysozyme signal peptide in the baker's yeast Saccharomyces cerevisiae expression host.The culture media of the bioengineered S.cerevisiae can degrade pure gliadin substrate and gliadin in wheat flour.The live yeast culture was demonstrated to degrade gliadin in wheat flour dough.The purified recombinant NrNpr1(rNrNpr1)enzyme was successfully characterized to have an optimal pH of 2.5,optimal temperature of 50℃,thermal tolerance of 0-65℃,and can digest all immunogenic epitopes inα-gliadin 33-mer when tested in vitro.ELISA test proved that rNrNpr1 can detoxify 750 mg/L gliadin to a gluten-free level(<20 mg/L)after 6 h of incubation.This study characterized neprosin from N.rafflesiana and report the bioengineering of S.cerevisiae with recombinant neprosin.We demonstrated that rNrNpr1 has promising potential applications in revolutionary gluten-safe food production,which needs further optimization and formulation in bakery.
文摘Research on marine bioactive peptides has mainly focused on characterization of peptides in hydrolysates prepared with commercial industrial enzymes and the usefulness of such hydrolysates in health and functional foods. However, a relevant question is whether digestion of fish proteins with gastrointestinal proteases per se generates peptides that also can have health promoting properties and can reduce, e.g., diabetes 2, inflammation and hypertension either in relation to gastrointestinal digestion or as alternative to industrial proteases. The aim of the study was to investigate hydrolysates obtained from in vitro sequential digestion of salmon muscle and skin with gastrointestinal proteases including pepsin, pancreatic and pancreatic ＋ mucosal proteases for their ability to scavenge ABTS^＋ radicals and inhibit activity of angiotensin I-converting enzyme （ACE） and dipeptidyl peptidase 4 （DPP-4）. Furthermore, it was the aim to study the inhibitory mechanism and stability towards ACE and DPP-4 activity. Analysis of〈 10 kDa hydrolysates showed that gastrointestinal proteases generated peptides with clear radical scavenging activity and DPP-4 and ACE inhibiting activity as well. Hydrolysates from pepsin digestion exhibited the lowest ECso values for radical scavenging activity and ACE inhibition, whereas ECso increased in hydrolysates after subsequent digestion with pancreatic and mucosal proteases. Interestingly, ECso values for the DPP-4 inhibition were hardly affected by sequential digestion. Inhibition modes for the muscle hydrolysates were both competitive and non-competitive, but prolonged incubation showed that the inhibitory properties were unstable and therefore they were probably digested as competitive substrates by gastrointestinal proteases.
文摘Objectives Dysregulated osteoclast function contributes to skeletal diseases.However,the specific ubiquitination regulators of the osteoclastogenesis repressor MafB,particularly at the post-translational level,remain undefined.This study aims to identify ubiquitin-specific proteases(USPs)that deubiquitinate MafB and enhance its stability.Methods We constructed a MafB-conjugated luciferase and overexpressed 40 individual USPs,measuring changes in luciferase activity.The identified USP was overexpressed in human CD14^(+) peripheral blood mononuclear cells(PBMCs)to evaluate its effect.Osteoclast differentiation was assessed through osteoclast marker Integrin alpha-V(CD51)staining and Western blot analysis.Co-immunoprecipitation(co-IP)was performed to assess the interplay.The influence on MafB ubiquitination and degradation was evaluated via immunoprecipitation and Western blot.Finally,MafB was knocked down in the USP-overexpressing PBMCs to analyze its effect on osteoclast differentiation.Results Overexpression of ubiquitin-specific protease 29(USP29)significantly increased MafB expression by approximately 75%(p<0.0001).Elevated USP29 levels strongly inhibited osteoclastic differentiation in CD14^(+) PBMCs(p<0.0001).USP29 was found to interact with MafB,markedly reducing its ubiquitination and subsequent degradation in PBMCs(p<0.001).Knocking down MafB in USP29-overexpressing PBMCs alleviated the inhibitory effect of USP29 on osteoclastogenesis.Conclusion USP29 acts as a potent stabilizer of MafB,inhibiting osteoclastogenesis in human CD14^(+) PBMCs,at least in part,by enhancing MafB stability.These findings expand our understanding of USP29’s role and the post-translational regulation of MafB.Furthermore,USP29 serves as a vital factor that controls osteoclast differentiation,and its regulatory function is at least partially mediated by deubiquitinating and stabilizing MafB.
