The main proteases(M^(pro)s)are hydrolases playing essential roles in the replication ofβ-coronaviruses including SARS-CoV-2.Herein,a highly efficient strategy was developed for discovering the M^(pro)inactivators fr...The main proteases(M^(pro)s)are hydrolases playing essential roles in the replication ofβ-coronaviruses including SARS-CoV-2.Herein,a highly efficient strategy was developed for discovering the M^(pro)inactivators from crude plant extract integrating target-based biochemical assay and chemoproteomic approaches.Firstly,Pu-erh tea was found to potently suppress SARS-CoV-2 M^(pro)in a time-dependent manner.Next,global chemical analysis coupling with peptide-modification profiling were used to identify the cysteine-modified constituents in Pu-erh tea.The results suggested that seven constituents in Pu-erh tea could modify SARSCoV-2 M^(pro),which turned out that epigallocatechin,gallocatechin and gallic acid were the most efficacious M^(pro)inactivators.Further investigations demonstrated that epigallocatechin and gallocatechin could inactivate S ARS-CoV-2 M^(pro)via blocking the formation of the homodimers.Collectively,this work proposed a novel and practical strategy for highly efficient discovery of time-dependent inhibitors of SARS-CoV-2 M^(pro)from plant extracts,while 3 constituents in Pu-erh tea have emerged as robust SARS-CoV-2 M^(pro)inactivators.展开更多
研究旨在探讨环二肽Cyclo(Leu-Pro)对植物乳杆菌生长的影响。该研究以植物乳杆菌(Lactobacillus Plantarum T1)为研究对象,采用高效液相色谱法初步定性定量植物乳杆菌T1发酵液中的Cyclo(Leu-Pro),然后将植物乳杆菌T1分别接种于不含或含...研究旨在探讨环二肽Cyclo(Leu-Pro)对植物乳杆菌生长的影响。该研究以植物乳杆菌(Lactobacillus Plantarum T1)为研究对象,采用高效液相色谱法初步定性定量植物乳杆菌T1发酵液中的Cyclo(Leu-Pro),然后将植物乳杆菌T1分别接种于不含或含不同质量浓度(0.5、1、5、10μg/mL)Cyclo(Leu-Pro)的MRS肉汤培养基中培养18 h,采用96孔法和平板计数法测定不同培养条件下植物乳杆菌T1的生长曲线和活菌数。结果表明:不同质量浓度的Cyclo(Leu-Pro)对植物乳杆菌T1的生长均有明显的促生长作用,其中,1μg/mL Cyclo(Leu-Pro)的促生长效果最佳,可将活菌数提高到8.73×10^(10) CFU/mL,相比于对照组提高了一个数量级。在不同胁迫条件下,1μg/mL Cyclo(Leu-Pro)能使植物乳杆菌T1的延滞期缩短、生长速度提高,其活菌数和生物量显著增加(P<0.05)。综上所述,Cyclo(Leu-Pro)促进了植物乳杆菌T1的生长,增强了其抗胁迫能力。这将有利于植物乳杆菌T1作为发酵剂在食品发酵领域的应用。展开更多
基金the supporting of Shanghai Science and Technology Innovation Action Plans(21S21900600,20S21901500,20S21900900)supported by Shanghai Science and Technology Committee+5 种基金the NSF of China(82273897,81922070,81973286,82104281,82173798)the Innovation Team and Talents Cultivation Program of National Administration of Traditional Chinese Medicine(ZYYCXTDD-202004)the Three-year Action Plan for Shanghai TCM Development and Inheritance Program(ZY(2021-2023)-0401)Shanghai Municipal Health Commission’s TCM Research Project(2022CX005)Shanghai Municipal Science and Technology Major Project(ZD2021CY001)Excellent Doctoral Student Cultivation Projects in Key Fields(GJ2022012)。
文摘The main proteases(M^(pro)s)are hydrolases playing essential roles in the replication ofβ-coronaviruses including SARS-CoV-2.Herein,a highly efficient strategy was developed for discovering the M^(pro)inactivators from crude plant extract integrating target-based biochemical assay and chemoproteomic approaches.Firstly,Pu-erh tea was found to potently suppress SARS-CoV-2 M^(pro)in a time-dependent manner.Next,global chemical analysis coupling with peptide-modification profiling were used to identify the cysteine-modified constituents in Pu-erh tea.The results suggested that seven constituents in Pu-erh tea could modify SARSCoV-2 M^(pro),which turned out that epigallocatechin,gallocatechin and gallic acid were the most efficacious M^(pro)inactivators.Further investigations demonstrated that epigallocatechin and gallocatechin could inactivate S ARS-CoV-2 M^(pro)via blocking the formation of the homodimers.Collectively,this work proposed a novel and practical strategy for highly efficient discovery of time-dependent inhibitors of SARS-CoV-2 M^(pro)from plant extracts,while 3 constituents in Pu-erh tea have emerged as robust SARS-CoV-2 M^(pro)inactivators.
