Characterization of SARS-COV-2 main protease inhibitory peptides from <italic style="font-style: italic">Ulva</italic> <italic style="font-style: italic">prolifera</italic> proteins

Zhiyong LI; Yehua WANG; Caiwei FU; Dongren ZHANG; Tuanjie CHE; Songdong SHEN

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Characterization of SARS-COV-2 main protease inhibitory peptides from Ulva prolifera proteins

Biology | Updated：2024-10-14

- Characterization of SARS-COV-2 main protease inhibitory peptides from Ulva prolifera proteins
- Journal of Oceanology and Limnology Vol. 41, Issue 5, Pages: 1887-1899(2023)
- Affiliations：
  
  1.School of Biology & Basic Medical Sciences, Soochow University, Suzhou 215101, China
  2.Key Laboratory of Functional Genomic and Molecular Diagnosis of Gansu Province, Lanzhou 730030, China
- Author bio：
  
  shensongdong@suda.edu.cn
- Funds：
- DOI：
  CLC：
- Received：27 April 2022，
  
  Accepted：09 June 2022，
  
  Online First：05 July 2022，
  
  Published：01 September 2023
- Accepted：
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LI Zhiyong,WANG Yehua,FU Caiwei,et al.Characterization of SARS-COV-2 main protease inhibitory peptides from Ulvaprolifera proteins[J].Journal of Oceanology and Limnology,2023,41(05):1887-1899.
DOI：

LI Zhiyong,WANG Yehua,FU Caiwei,et al.Characterization of SARS-COV-2 main protease inhibitory peptides from Ulvaprolifera proteins[J].Journal of Oceanology and Limnology,2023,41(05):1887-1899. DOI：

摘要

Abstract

The main protease (M

pro

) is essential for the replication of SARS-COV-2 and therefore represents a promising anti-viral target. In this study

we screened M

pro

inhibitory peptides from

Ulva

prolifera

protein on in-silico proteolysis. Cytotoxicity analysis using the online toxic prediction tool ToxinPred revealed that all the peptides were non-cytotoxic. The hexapeptide (SSGFID) exhibited high M

pro

inhibitory activity in molecular docking and its IC

value was 139.40±0.82 μmol/L in vitro according to fluorescence resonance energy transfer assay (FRET). Quantitative real-time (qRT-) PCR results show that SSGFID could stimulate the expression of mitosis-related factors

including nuclear factor-κB

cyclin D1

and cyclin-dependent kinase 4

to promote the proliferation of mice splenocytes. Stability study revealed that SSGFID showed resistance against pepsin and trypsin but lost D (Asp) after pretreatment at 121 °‍‍C for 15 min. Besides

SSGFID was mainly transported through the Caco-2 cell monolayer by the peptide transporter PepT1 and passive-mediated transport during the transport study. Unfortunately

the peptide was also degraded by Caco-2 intracellular enzymes

and the transfer rate of intact peptide was 4.2%. Furthermore

Lineweaver–Burk plots demonstrated that SSGFID possessed a mixed inhibitory characteristic with M

pro

. Our study indicated the potential of

Ulva

prolifera

as antiviral and immune-enhancing functional food ingredients and nutraceuticals.

关键词

Keywords

references

Abuo-Rahma G E D A , Mohamed M F A , Ibrahim T S et al . 2020 . Potential repurposed SARS-CoV-2 (COVID-19) infection drugs . RSC Advances , 10 ( 45 ): 26895 - 26916 , https://doi.org/10.1039/D0RA05821A https://doi.org/10.1039/D0RA05821A .

Aguilar-Briseño J A , Cruz-Suarez L E , Sassi J F et al . 2015 . Sulphated polysaccharides from Ulva clathrata and Cladosiphon okamuranus seaweeds both inhibit viral attachment/entry and cell-cell fusion, in NDV infection . Marine Drugs , 13 ( 2 ): 697 - 712 , https://doi.org/10.3390/md13020697 https://doi.org/10.3390/md13020697 .

Aguilera-Morales M , Casas-Valdez M , Carrillo-Domı́nguez S et al . 2005 . Chemical composition and microbiological assays of marine algae Enteromorpha spp. as a potential food source . Journal of Food Composition and Analysis , 18 ( 1 ): 79 - 88 , https://doi.org/10.1016/j.jfca.2003.12.012 https://doi.org/10.1016/j.jfca.2003.12.012 .

Ahn G , Hwang I , Park E et al . 2008 . Immunomodulatory effects of an enzymatic extract from Ecklonia cava on murine splenocytes . Marine Biotechnology , 10 ( 3 ): 278 - 289 , https://doi.org/10.1007/s10126-007-9062-9 https://doi.org/10.1007/s10126-007-9062-9 .

Alanyali S , Yeniay L , Demir D et al . 2017 . Expression of Cyclin D1 and its relationship to hormone receptors and survival in breast cancer . Turkish Journal of Oncology , 32 ( 1 ): 7 - 13 , https://doi.org/10.5505/tjo.2017.1386 https://doi.org/10.5505/tjo.2017.1386 .

Artursson P , Palm K , Luthman K . 1996 . Caco-2 monolayers in experimental and theoretical predictions of drug transport . Advanced Drug Delivery Reviews , 22 ( 1-2 ): 67 - 84 , https://doi.org/10.1016/S0169-409X(96)00415-2 https://doi.org/10.1016/S0169-409X(96)00415-2 .

Chiu Y H , Chan Y L , Li T L et al . 2012 . Inhibition of Japanese Encephalitis virus infection by the sulfated polysaccharide extracts from Ulva lactuca . Marine Biotechnology , 14 ( 4 ): 468 - 478 , https://doi.org/10.1007/s10126-011-9428-x https://doi.org/10.1007/s10126-011-9428-x .

Cian R E , López-Posadas R , Drago S R et al . 2012 . A Porphyra columbina hydrolysate upregulates IL-10 production in rat macrophages and lymphocytes through an NF-κB, and p38 and JNK dependent mechanism . Food Chemistry , 134 ( 4 ): 1982 - 1990 , https://doi.org/10.1016/j.foodchem.2012.03.134 https://doi.org/10.1016/j.foodchem.2012.03.134 .

Dai W H , Zhang B , Jiang X M et al . 2020 . Structure-based design of antiviral drug candidates targeting the SARS-CoV-2 main protease . Science , 368 ( 6497 ): 1331 - 1335 , https://doi.org/10.1126/science.abb4489 https://doi.org/10.1126/science.abb4489 .

Ding X M , Hu X Y , Chen Y et al . 2021 . Differentiated Caco-2 cell models in food-intestine interaction study: current applications and future trends . Trends in Food Science & Technology , 107 : 455 - 465 , https://doi.org/10.1016/j.tifs.2020.11.015 https://doi.org/10.1016/j.tifs.2020.11.015 .

Drugs . com . 2023 . Paxlovid Side Effects from Drugs. com. Drugs , 18 March 2023, https://www.drugs.com/sfx/paxlovid-side-effects.html. Accessed on 2023-03-18 https://www.drugs.com/sfx/paxlovid-side-effects.html.Accessedon2023-03-18 .

Engel M , Hoffmann T , Wagner L et al . 2003 . The crystal structure of dipeptidyl peptidase IV (CD26) reveals its functional regulation and enzymatic mechanism . Proceedings of the National Academy of Sciences of the United States of America , 100 ( 9 ): 5063 - 5068 , https://doi.org/10.1073/pnas.0230620100 https://doi.org/10.1073/pnas.0230620100 .

Fan H B , Wu J P . 2021 . Purification and identification of novel ACE inhibitory and ACE2 upregulating peptides from spent hen muscle proteins . Food Chemistry , 345 : 128867 , https://doi.org/10.1016/j.foodchem.2020.128867 https://doi.org/10.1016/j.foodchem.2020.128867 .

Fan H B , Xu Q B , Hong H et al . 2018 . Stability and transport of spent hen-derived ACE-inhibitory peptides IWHHT, IWH, and IW in human intestinal Caco-2 cell monolayers . Journal of Agricultural and Food Chemistry , 66 ( 43 ): 11347 - 11354 , https://doi.org/10.1021/acs.jafc.8b03956 https://doi.org/10.1021/acs.jafc.8b03956 .

Gentile D , Patamia V , Scala A et al . 2020 . Putative inhibitors of SARS-CoV-2 main protease from a library of marine natural products: a virtual screening and molecular modeling study . Marine Drugs , 18 ( 4 ): 225 . https://do 10.3390/md18040225 http://dx.doi.org/10.3390/md18040225

Guo C , Li F C , Jiang P et al . 2011 . Bacterial diversity in surface water of the Yellow Sea during and after a green alga tide in 2008 . Chinese Journal of Oceanology and Limnology , 29 ( 6 ): 1147 - 1154 , https://doi.org/10.1007/s00343-011-0264-7 https://doi.org/10.1007/s00343-011-0264-7 .

Han H K , de Vrueh R L A , Rhie J K et al . 1998 . 5′-Amino acid esters of antiviral nucleosides, acyclovir, and AZT are absorbed by the intestinal PEPT1 peptide transporter . Pharmaceutical Research , 15 ( 8 ): 1154 - 1159 , https://doi.org/10.1023/A:1011919319810 https://doi.org/10.1023/A:1011919319810 .

Hardouin K , Bedoux G , Burlot A S et al . 2016 . Enzyme-assisted extraction (EAE) for the production of antiviral and antioxidant extracts from the green seaweed Ulva armoricana (Ulvales, Ulvophyceae) . Algal Research , 16 : 233 - 239 , https://doi.org/10.1016/j.algal.2016.03.013 https://doi.org/10.1016/j.algal.2016.03.013 .

Henriques S T , Huang Y H , Rosengren K J et al . 2011 . Decoding the membrane activity of the cyclotide kalata B1: the importance of phosphatidylethanolamine phospholipids and lipid organization on hemolytic and anti-HIV activities . Journal of Biological Chemistry , 286 : 24231 - 24241 , https://doi.org/10.1074/jbc.M111.253393 https://doi.org/10.1074/jbc.M111.253393 .

Hong W , Li T , Song Y et al . 2014 . Inhibitory activity and mechanism of two scorpion venom peptides against herpes simplex virus type 1 . Antiviral Research , 102 : 1 - 10 , https://doi.org/10.1016/j.antiviral.2013.11.013 https://doi.org/10.1016/j.antiviral.2013.11.013 .

Hooton D , Lentle R , Monro J et al . 2015 . The secretion and action of brush border enzymes in the mammalian small intestine . In: Nilius B, Gudermann T, Jahn R et al. eds . Reviews of Physiology, Biochemistry and Pharmacology. Springer, Cham . p. 59 - 118 , https://doi.‍org/10.1007/112_2015_24 https://doi.‍org/10.1007/112_2015_24 . https://do 10.1007/112_2015_24 http://dx.doi.org/10.1007/112_2015_24

Hou H , Fan Y , Li B F et al . 2012 . Purification and identification of immunomodulating peptides from enzymatic hydrolysates of Alaska pollock frame . Food Chemistry , 134 ( 2 ): 821 - 828 , https://doi.org/10.1016/j.foodchem.2012.02.186 https://doi.org/10.1016/j.foodchem.2012.02.186 .

Ibrahim M A A , Abdelrahman A H M , Hussien T A et al . 2020 . In silico drug discovery of major metabolites from spices as SARS-CoV-2 main protease inhibitors . Computers in Biology and Medicine , 126 : 104046 , https://doi.org/10.1016/j.compbiomed.2020.104046 https://doi.org/10.1016/j.compbiomed.2020.104046 .

Jin Z M , Du X Y , Xu Y C et al . 2020 . Structure of M pro from SARS-CoV-2 and discovery of its inhibitors . Nature , 582 ( 7811 ): 289 - 293 , https://doi.org/10.1038/s41586-020-2223-y https://doi.org/10.1038/s41586-020-2223-y .

Kim Y , Liu H W , Kankanamalage A C G et al . 2016 . Correction: reversal of the progression of fatal coronavirus infection in cats by a broad-spectrum coronavirus protease inhibitor . PLoS Pathogens , 12 ( 5 ): e1005650 , https://doi.org/10.1371/journal.ppat.1005650 https://doi.org/10.1371/journal.ppat.1005650 .

Lacroix I M , Chen X M , Kitts D D et al . 2017 . Investigation into the bioavailability of milk protein-derived peptides with dipeptidyl-peptidase IV inhibitory activity using Caco-2 cell monolayers . Food & Function , 8 ( 2 ): 701 - 709 , https://doi.org/10.1039/C6FO01411A https://doi.org/10.1039/C6FO01411A .

Landowski C P , Song X Q , Lorenzi P L et al . 2005 . Floxuridine amino acid ester prodrugs: enhancing Caco-2 permeability and resistance to glycosidic bond metabolism . Pharmaceutical Research , 22 ( 9 ): 1510 - 1518 , https://doi.org/10.1007/s11095-005-6156-9 https://doi.org/10.1007/s11095-005-6156-9

Lehmann T , Köhler C , Weidauer E et al . 2001 . Expression of MRP1 and related transporters in human lung cells in culture . Toxicology , 167 ( 1 ): 59 - 72 , https://doi.org/10.1016/S0300-483X(01)00458-9 https://doi.org/10.1016/S0300-483X(01)00458-9 .

Li H M , Zhang Y Y , Han X R et al . 2016 . Growth responses of Ulva prolifera to inorganic and organic nutrients: implications for macroalgal blooms in the southern Yellow Sea, China . Scientific Reports , 6 : 26498 , https://doi.org/10.1038/srep26498 https://doi.org/10.1038/srep26498 .

Li Z B , Yang H , Shangguan J et al . 2018 . Growth performance, digestive enzyme activities and serum nonspecific immunity of the red tilapia ( Oreochromis mossambicus × Oreochromis niloticus ) fed diets supplemented with ultrafine powder of Enteromopha prolifera . Journal of Oceanology and Limnology , 36 ( 5 ): 1843 - 1850 , https://doi.org/10.1007/s00343-018-7142-5 https://doi.org/10.1007/s00343-018-7142-5 .

Li Z Y , Zhao S , Xin X D et al . 2020 . Purification, identification and functional analysis of a novel immunomodulatory peptide from silkworm pupa protein . International Journal of Peptide Research and Therapeutics , 26 ( 1 ): 243 - 249 , https://doi.org/10.1007/s10989-019-09832-4 https://doi.org/10.1007/s10989-019-09832-4 .

Liu P , Liao W , Qi X P et al . 2020 . Identification of immunomodulatory peptides from zein hydrolysates . European Food Research and Technology , 246 ( 5 ): 931 - 937 , https://doi.org/10.1007/s00217-020-03450-x https://doi.org/10.1007/s00217-020-03450-x .

Ma C L , Sacco M D , Hurst B et al . 2020 . Boceprevir, GC-376, and calpain inhibitors II, XII inhibit SARS-CoV-2 viral replication by targeting the viral main protease . Cell Research , 30 ( 8 ): 678 - 692 , https://doi.org/10.1038/s41422-020-0356-z https://doi.org/10.1038/s41422-020-0356-z .

McKimm-Breschkin J L , Hay A J , Cao B et al . 2022 . COVID-19, Influenza and RSV: surveillance-informed prevention and treatment—Meeting report from an isirv-WHO virtual conference . Antiviral Research , 197 : 105227 , https://doi.org/10.1016/j.antiviral.2021.105227 https://doi.org/10.1016/j.antiviral.2021.105227 .

Morris H J , Carrillo O , Almarales A et al . 2007 . Immunostimulant activity of an enzymatic protein hydrolysate from green microalga Chlorella vulgaris on undernourished mice . Enzyme and Microbial Technology , 40 ( 3 ): 456 - 460 , https://doi.org/10.1016/j.enzmictec.2006.07.021 https://doi.org/10.1016/j.enzmictec.2006.07.021 .

Owen D R , Allerton C M N , Anderson A S et al . 2021 . An oral SARS-CoV-2 M pro inhibitor clinical candidate for the treatment of COVID-19 . Science , 374 ( 6575 ): 1586 - 1593 , https://doi.org/10.1126/science.abl4784 https://doi.org/10.1126/science.abl4784 .

Pan S , Wang S , Jing L et al . 2016 . Purification and characterisation of a novel angiotensin-I converting enzy me (ACE)-inhibitory peptide derived from the enzymatic hydrolysate of Enteromorpha clathrata protein . Food Chemistry , 211 : 423 - 430 , https://doi.org/10.1016/j.foodchem.2016.05.087 https://doi.org/10.1016/j.foodchem.2016.05.087 .

Pan S K , Wang S J , Jing L L et al . 2016 . Purification and characterisation of a novel angiotensin-I converting enzyme (ACE)-inhibitory peptide derived from the enzymatic hydrolysate of Enteromorpha clathrata protein . Food Chemistry , 211 : 423 - 430 , https://doi.org/10.1016/j.foodchem.2016.05.087 https://doi.org/10.1016/j.foodchem.2016.05.087 .

Pappenheimer J R , Dahl C E , Karnovsky M L et al . 1994 . Intestinal absorption and excretion of octapeptidescomposed of D amino acids . Proceedings of the National Academy of Sciences of the United States of America , 91 ( 5 ): 1942 - 1945 , https://doi.org/10.1073/pnas.91.5.1942 https://doi.org/10.1073/pnas.91.5.1942 .

Pei J Y , Hua Y , Zhou T Y et al . 2021 . Transport, in vivo antihypertensive effect, and pharmacokinetics of an Angiotensin-Converting Enzyme (ACE) inhibitory peptide LVLPGE . Journal of Agricultural and Food Chemistry , 69 ( 7 ): 2149 - 2156 , https://doi.org/10.1021/acs.jafc.0c07048 https://doi.org/10.1021/acs.jafc.0c07048 .

Ramajayam R , Tan K P , Liang P H . 2011 . Recent development of 3C and 3CL protease inhibitors for anti-coronavirus and anti-picornavirus drug discovery . Biochemical Society Transactions , 39 ( 5 ): 1371 - 1375 , https://doi.org/10.1042/BST0391371 https://doi.org/10.1042/BST0391371 .

Rothan H A , Bahrani H , Rahman N A et al . 2014 . Identification of natural antimicrobial agents to treat dengue infection: in vitro analysis of latarcin peptide activity against dengue virus . BMC Microbiology , 14 : 140 , https://doi.org/10.1186/1471-2180-14-140 https://doi.org/10.1186/1471-2180-14-140 .

Santiago-López L , Hernández-Mendoza A , Vallejo-Cordoba B et al . 2016 . Food-derived immunomodulatory peptides . Journal of the Science of Food and Agriculture , 96 ( 11 ): 3631 - 3641 , https://doi.org/10.1002/jsfa.7697 https://doi.org/10.1002/jsfa.7697 .

Satake M , Enjoh M , Nakamura Y et al . 2002 . Transepithelial transport of the bioactive tripeptide, Val-Pro-Pro, in human intestinal Caco-2 cell monolayers . Bioscience, Biotechnology, and Biochemistry , 66 ( 2 ): 378 - 384 , https://doi.org/10.1271/bbb.66.378 https://doi.org/10.1271/bbb.66.378 .

Sütas Y , Soppi E , Korhonen H et al . 1996 . Suppression of lymphocyte proliferation in vitro by bovine caseins hydrolyzed with Lactobacillus casei GG-derived enzymes . Journal of Allergy and Clinical Immunology , 98 ( 1 ): 216 - 224 , https://doi.org/10.1016/S0091-6749(96)70245-2 https://doi.org/10.1016/S0091-6749(96)70245-2 .

Sun Y , Liu J , Xia J et al . 2022 . Research development on resource utilization of green tide algae from the Southern Yellow Sea . Energy Reports , 8 : 295 - 303 , https://doi.org/10.1016/j.egyr.2022.01.168 https://doi.org/10.1016/j.egyr.2022.01.168 .

Tamura K , Bhatnagar P K , Takata J S et al . 1996 . Metabolism, uptake, and transepithelial transport of the diastereomers of Val-Val in the human intestinal cell line, Caco-2 . Pharmaceutical Research , 13 ( 8 ): 1213 - 1218 , https://doi.org/10.1023/A:1016068421243 https://doi.org/10.1023/A:1016068421243 .

Tu M L , Liu H X , Zhang R Y et al . 2018 . Analysis and evaluation of the inhibitory mechanism of a novel angiotensin-I-converting enzyme inhibitory peptide derived from casein hydrolysate . Journal of Agricultural and Food Chemistry , 66 ( 16 ): 4139 - 4144 , https://doi.org/10.1021/acs.jafc.8b00732 https://doi.org/10.1021/acs.jafc.8b00732 .

Udenigwe C C . 2014 . Bioinformatics approaches, prospects and challenges of food bioactive peptide research . Trends in Food Science & Technology , 36 ( 2 ): 137 - 143 , https://doi.org/10.1016/j.tifs.2014.02.004 https://doi.org/10.1016/j.tifs.2014.02.004 .

Vijayakumar M , Janani B , Kannappan P et al . 2022 . In silico identification of potential inhibitors against main protease of SARS-CoV-2 6LU7 from Andrographis panniculata via molecular docking, binding energy calculations and molecular dynamics simulation studies . Saudi Journal of Biological Sciences , 29 ( 1 ): 18 - 29 , https://doi.org/10.1016/j.sjbs.2021.10.060 https://doi.org/10.1016/j.sjbs.2021.10.060 .

Wachinger M , Kleinschmidt A , Winder D et al . 1998 . Antimicrobial peptides melittin and cecropin inhibit replication of human immunodeficiency virus 1 by suppressing viral gene expression . Journal of General Virology , 79 ( 4 ): 731 - 740 , https://doi.org/10.1099/0022-1317-79-4-731 https://doi.org/10.1099/0022-1317-79-4-731 .

Xu Z Q , Wu C P , Sun-Waterhouse D et al . 2021 . Identification of post-digestion angiotensin-I converting enzyme (ACE) inhibitory peptides from soybean protein Isolate: their production conditions and in silico molecular docking with ACE . Food Chemistry , 345 : 128855 , https://doi.org/10.1016/j.foodchem.2020.128855 https://doi.org/10.1016/j.foodchem.2020.128855 .

Yamauchi F , Suetsuna K . 1993 . Immunological effects of dietary peptide derived from soybean protein . The Journal of Nutritional Biochemistry , 4 ( 8 ): 450 - 457 , https://doi.org/10.1016/0955-2863(93)90062-2 https://doi.org/10.1016/0955-2863(93)90062-2 .

Yang Q , Cai X X , Huang M C et al . 2020 . A specific peptide with immunomodulatory activity from Pseudostellaria heterophylla and the action mechanism . Journal of Functional Foods , 68 : 103887 , https://doi.org/10.1016/j.jff.2020.103887 https://doi.org/10.1016/j.jff.2020.103887 .

Yang R Y , Zhang Z F , Pei X R et al . 2009 . Immunomodulatory effects of marine oligopeptide preparation from Chum Salmon ( Oncorhynchus keta ) in mice . Food Chemistry , 113 ( 2 ): 464 - 470 , https://doi.org/10.1016/j.foodchem.2008.07.086 https://doi.org/10.1016/j.foodchem.2008.07.086 .

Ye N H , Zhang X W , Mao Y Z et al . 2011 . 'Green tides' are overwhelming the coastline of our blue planet: taking the world's largest example . Ecological Research , 26 ( 3 ): 477 - 485 , https://doi.org/10.1007/s11284-011-0821-8 https://doi.org/10.1007/s11284-011-0821-8 .

Zhang L L , Lin D Z , Kusov Y et al . 2020 . α-Ketoamides as broad-spectrum inhibitors of coronavirus and enterovirus replication: structure-based design, synthesis, and activity assessment . Journal of Medicinal Chemistry , 63 ( 9 ): 4562 - 4578 , https://doi.org/10.1021/acs.jmedchem.9b01828 https://doi.org/10.1021/acs.jmedchem.9b01828 .

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Characterization of SARS-COV-2 main protease inhibitory peptides from Ulva prolifera proteins

DOI：

摘要

Abstract

关键词

Keywords

references