Abstract
Protein hydrolysates are easily digested and utilized by humans and animals, and are less likely to cause allergies. Protein hydrolysis caused by endopeptidases often leads to the exposure of hydrophobic amino acids at the ends of peptides, which consequently causes bitter taste. Microbial aminopeptidases remove the exposed hydrophobic amino acids at the ends of aminopeptides, which improves taste, allowing for easier production. This processe is attacking significant attention from industry and laboratories. Aminopeptidases selectively hydrolyze peptide bonds from the N-terminal of proteins or peptides to produce free amino acids. Aminopeptidases can be classified into leucine, lysine, methionine and proline aminopeptidases by hydrolyzed N-terminal residues; metallo-, serine- and cysteine- aminopeptidases by the reaction mechanisms; dipeptide and triphoptide enzymes by the released number of amino acid residues at the end of hydrolyzed peptides; or acidic, neutral and basic aminopeptidases by their optimal hydrolysis pH. Commercial aminopeptidases are generally produced by microbial fermentation, and are mainly applied in the debittering of protein hydrolysates, the deep hydrolysis of protein, and the production of condiments, cheese, and bioactive peptides, as well as for disease detection in the medical industry.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
All data generated or analyzed during this study are included in this published article.
References
Are VN, Kumar A, Goyal VD et al (2019) Structures and activities of widely conserved small prokaryotic aminopeptidases-P clarify classification of M24B peptidases. Proteins 87(3):212–225. https://doi.org/10.1002/prot.25641
Arima J, Uesugi Y, Iwabuchi M et al (2006a) Study on peptide hydrolysis by aminopeptidases from Streptomyces griseus, Streptomyces septatus and Aeromonas proteolytica. Appl Microbiol Biotechnol 70:541–547. https://doi.org/10.1007/s00253-005-0105-8
Arima J, Uesugi Y, Uraji M et al (2006b) Dipeptide synthesis by an aminopeptidase from Streptomyces septatus TH-2 and its application to synthesis of biologically active peptides. Appl Environ Microbiol 72(6):4225–4231. https://doi.org/10.1128/AEM.00150-06
Arima J, Morimoto M, Usuki H et al (2010) Beta-alanyl peptide synthesis by Streptomyces S9 aminopeptidase. J Biotechnol 147(1):52–58. https://doi.org/10.1016/j.jbiotec.2010.03.007
Arima J, Tokai S, Chiba M et al (2014) Gene cloning and biochemical characterization of eryngase, a serine aminopeptidase of Pleurotus eryngii belonging to the family S9 peptidases. Biosci Biotech Biochem 78(11):1856–1863. https://doi.org/10.1080/09168451.2014.940277
Arya PS, Yagnik SM, Rajput KN et al (2021) Understanding the basis of occurrence, biosynthesis, and implications of thermostable alkaline proteases. Appl Biochem Biotech 193(12):4113–4150. https://doi.org/10.1007/s12010-021-03701-x
Axelrad I, Safrin M, Cahan R et al (2021) Extracellular proteolytic activation of Pseudomonas aeruginosa aminopeptidase (PaAP) and insight into the role of its non-catalytic N-terminal domain. PLoS ONE 16(6):e0252970. https://doi.org/10.1371/journal.pone.0252970
Bocanegra-Jiménez FY, Montero-Morán GM, Lara-González S (2021) Purification and characterization of an FeII - and α-ketoglutarate-dependent xanthine hydroxylase from Aspergillus oryzae. Protein Expres Purif 183:105862. https://doi.org/10.1016/j.pep.2021.105862
Chen Y, Zhang R, Zhang W et al (2022) Alanine aminopeptidase from Bacillus licheniformis E7 expressed in Bacillus subtilis efficiently hydrolyzes soy protein to small peptides and free amino acids. LWT-Food Sci Technol 165:113642. https://doi.org/10.1016/j.lwt.2022.113642
de Diego I, Veillard FT, Guevara T et al (2013) Porphyromonas gingivalis virulence factor gingipain RgpB shows a unique zymogenic mechanism for cysteine peptidases. J Biol Chem 288(20):14287–14296. https://doi.org/10.1074/jbc.M112.444927
de Palencia PF, de Felipe FL, Requena T et al (2000) The aminopeptidase C (PepC) from Lactobacillus helveticus CNRZ32. A comparative study of PepC from lactic acid bacteria. Eur Food Res Technol 212(1):89–94. https://doi.org/10.1007/s002170000203
Dong Z, Yang S, Zhang Z et al (2022) Prolyl aminopeptidases: reclassification, properties, production and industrial applications. Process Biochem 118:121–132. https://doi.org/10.1016/j.procbio.2022.04.025
Ewert J, Schlierenkamp F, Nesensohn L et al (2018) Improving the colloidal and sensory properties of a caseinate hydrolysate using particular exopeptidases. Food Funct 9(11):5989–5998. https://doi.org/10.1039/c8fo01749b
Fu Y, Chen J, Bak KH et al (2019) Valorisation of protein hydrolysates from animal by-products: perspectives on bitter taste and debittering methods: a review. Int J Food Sci Tech 54(4):978–986. https://doi.org/10.1111/ijfs.14037
Fundoiano-Hershcovitz Y, Rabinovitch L, Shulami S et al (2005) The ywad gene from Bacillus subtilis encodes a double-zinc aminopeptidase. FEMS Microbiol Lett 243(1):157–163. https://doi.org/10.1016/j.femsle.2004.12.001
Gao X, Cui C, Ren J et al (2011) Changes in the chemical composition of traditional Chinese-type soy sauce at different stages of manufacture and its relation to taste. Int J Food Sci Tech 46(2):243–249. https://doi.org/10.1111/j.1365-2621.2010.02487.x
Gao X, Cui W, Ding N et al (2013) Structure-based approach to alter the substrate specificity of Bacillus subtilis aminopeptidase. Prion 7(4):328–334. https://doi.org/10.4161/pri.25147
Gao X, Yin Y, Zhou C (2018) Purification, characterisation and salt-tolerance molecular mechanisms of aspartyl aminopeptidase from Aspergillus oryzae 3.042. Food Chem 240:377–385. https://doi.org/10.1016/j.foodchem.2017.07.081
González-Bacerio J, Varela AC, Aguado ME et al (2022) Bacterial metalo-aminopeptidases as targets in human infectious diseases. Curr Drug Targets 23(12):1155–1190. https://doi.org/10.2174/1389450123666220316085859
Guleria S, Walia A, Chauhan A et al (2016) Molecular characterization of alkaline protease of Bacillus amyloliquefaciens SP1 involved in biocontrol of Fusarium oxysporum. Int J Food Microbiol 232:134–143. https://doi.org/10.1016/j.ijfoodmicro.2016.05.030
Hakim A, Bhuiyan FR, Iqbal A et al (2018) Production and partial characterization of dehairing alkaline protease from Bacillus subtilis AKAL7 and Exiguobacterium indicum AKAL11 by using organic municipal solid wastes. Heliyon 4(6):e00646. https://doi.org/10.1016/j.heliyon.2018.e00646
Han Y, Guo C, Yan Z et al (2018) A proteome-based design of bitter peptide digestion regime to attenuate bone soup bitterness: comparison with a rainbow trout extract-mediated bitter taste masking approach. bioRxiv. https://doi.org/10.1101/279265
Holz RC (2002) The aminopeptidase from Aeromonas proteolytica: structure and mechanism of co-catalytic metal centers involved in peptide hydrolysis. Coordin Chem Rev 232(1):5–26. https://doi.org/10.1016/S0010-8545(01)00470-2
Hu X, Zhang Q, Zhang Q et al (2022) An updated review of functional properties, debittering methods, and applications of soybean functional peptides. Crit Rev Food Sci 321:1–16. https://doi.org/10.1080/10408398.2022.2062587
Idowu AT, Benjakul S (2019) Bitterness of fish protein hydrolysate and its debittering prospects. J Food Biochem. https://doi.org/10.1111/jfbc.12978
Isshiki S, Sachiyo S, Shouko K et al (2017) Characterization of an aminopeptidase from Pseudozyma hubeiensis 31-B and potential applications. Mycoscience 58(1):60–67. https://doi.org/10.1016/j.myc.2016.10.001
Ito K, Nakajima Y, Onohara Y et al (2006) Crystal structure of aminopeptidase N (proteobacteria alanyl aminopeptidase) from Escherichia coli and conformational change of methionine 260 involved in substrate recognition. J Biol Chem 281(44):33664–33676. https://doi.org/10.1074/jbc.M605203200
Kiehstaller S, Ottmann C, Hennig S (2020) MMP activation–associated aminopeptidase N reveals a bivalent 14–3-3 binding motif. J Biol Chem 295(52):18266–18275. https://doi.org/10.1074/jbc.RA120.014708
Kim JH, Lee BR, Lee YP (2011) Secretory overproduction of the aminopeptidase from Bacillus licheniformis by a novel hybrid promoter in Bacillus subtilis. World J Microbiol Biotechnol 27(11):2747–2751. https://doi.org/10.1007/s11274-011-0749-8
Kremer A, Li SM (2008) Tryptophan aminopeptidase activity of several indole prenyltransferases from Aspergillus fumigatus. Chem Biol 15(7):729–738. https://doi.org/10.1016/j.chembiol.2008.05.019
Laskar A, Chatterjee A, Chatterjee S et al (2012) Three-dimensional molecular modeling of a diverse range of SC clan serine proteases. Mol Biol Int 2012:580965. https://doi.org/10.1155/2012/580965
Lei F, Zhao Q, Sun-Waterhouse D et al (2017) Characterization of a salt-tolerant aminopeptidase from marine Bacillus licheniformis SWJS33 that improves hydrolysis and debittering efficiency for soy protein isolate. Food Chem 214:347–353. https://doi.org/10.1016/j.foodchem.2016.07.028
Lei F, Chuanrong H, Dongping H et al (2019) The specificity of an aminopeptidase affects its performance in hydrolyzing peanut protein isolate and zein. LWT-Food Sci Technol 102:37–44. https://doi.org/10.1016/j.lwt.2018.10.041
Li L, Yang ZY, Yang XQ et al (2008) Debittering effect of Actinomucor elegans peptidases on soybean protein hydrolysates. J Ind Microbiol Biotechnol 35(1):41–47. https://doi.org/10.1007/s10295-007-0264-y
Lin X, Dong L, Yu D et al (2020) High-level expression and characterization of the thermostable leucine aminopeptidase Thelap from the thermophilic fungus Thermomyces lanuginosus in Aspergillus niger and its application in soy protein hydrolysis. Protein Expres Purif 167:105544. https://doi.org/10.1016/j.pep.2019.105544
Liu X, Jiang D, Peterson DG (2014) Identification of bitter peptides in whey protein hydrolysate. J Agr Food Chem 62(25):5719–5725. https://doi.org/10.1021/jf4019728
Majumdar S, Sarmah B, Gogoi D et al (2014) Characterization, mechanism of anticoagulant action, and assessment of therapeutic potential of a fibrinolytic serine protease (Brevithrombolase) purified from Brevibacillus brevis strain FF02B. Biochimie 103:50–60. https://doi.org/10.1016/j.biochi.2014.04.002
Malcolm TR, Swiderska KW, Hayes BK et al (2021) Mapping the substrate specificity of the Plasmodium M1 and M17 aminopeptidases. Biochem J 478(13):2697–2713. https://doi.org/10.1042/BCJ20210172
Marusic N, Vidacek S, Janci T et al (2014) Determination of volatile compounds and quality parameters of traditional Istrian dry-cured ham. Meat Sci 96(4):1409–1416. https://doi.org/10.1016/j.meatsci.2013.12.003
Matkawala F, Nighojkar S, Kumar A et al (2021) Microbial alkaline serine proteases: production, properties and applications. World J Microbiol Biotechnol 37(4):63. https://doi.org/10.1007/s11274-021-03036-z
Matsushita-Morita M, Tada S, Suzuki S et al (2011) Overexpression and characterization of an extracellular leucine aminopeptidase from Aspergillus oryzae. Curr Microbiol 62(2):557–564. https://doi.org/10.1007/s00284-010-9744-9
Méndez Y, Pérez-Labrada K, González-Bacerio J et al (2014) Combinatorial multicomponent access to natural-products-inspired peptidomimetics: discovery of selective inhibitors of microbial metallo-aminopeptidases. Chem Med Chem 9(10):2351–2359. https://doi.org/10.1002/cmdc.201402140
Mika N, Zorn H, Rühl M (2015) Prolyl-specific peptidases for applications in food protein hydrolysis. Appl Microbiol Biotechnol 99(19):7837–7846. https://doi.org/10.1007/s00253-015-6838-0
Minasov G, Lam MR, Rosas-Lemus M et al (2020) Comparison of metal-bound and unbound structures of aminopeptidase B proteins from Escherichia coli and Yersinia pestis. Protein Sci 29(7):1618–1628. https://doi.org/10.1002/pro.3876
Modak JK, Rut W, Wijeyewickrema LC et al (2016) Structural basis for substrate specificity of Helicobacter pylori M17 aminopeptidase. Biochimie 121:60–71. https://doi.org/10.1016/j.biochi.2015.11.021
Nagendra Kumar C, Deepali K, Anjali K et al (2022) Proteolytic enzymes derived from a macro fungus and their industrial application. Hydrolases. IntechOpen, London
Nakajima Y, Ito K, Sakata M et al (2006) Unusual extra space at the active site and high activity for acetylated hydroxyproline of prolyl aminopeptidase from Serratia marcescens. J Bacteriol 188(4):1599–1606. https://doi.org/10.1128/JB.188.4.1599-1606.2006
Nandan A, Nampoothiri KM (2017) Molecular advances in microbial aminopeptidases. Bioresour Technol 245:1757–1765. https://doi.org/10.1016/j.biortech.2017.05.103
Nandan A, Nampoothiri KM (2020) Therapeutic and biotechnological applications of substrate specific microbial aminopeptidases. Appl Microbiol Biotechnol 104(12):5243–5257. https://doi.org/10.1007/s00253-020-10641-9
Naveed M, Nadeem F, Mehmood T et al (2021) Protease—a versatile and ecofriendly biocatalyst with multi-industrial applications: an updated review. Catal Lett 151(2):307–323. https://doi.org/10.1007/s10562-020-03316-7
Nishiwaki T, Yoshimizu S, Furuta M et al (2002) Debittering of enzymatic hydrolysates using an aminopeptidase from the edible basidiomycete Grifola frondosa. J Biosci Bioeng 93(1):60–63. https://doi.org/10.1016/S1389-1723(02)80055-X
Page MJ, Di Cera E (2008) Serine peptidases: classification, structure and function. Cell Mol Life Sci 65(7–8):1220–1236. https://doi.org/10.1007/s00018-008-7565-9
Pérez-Palacios T, Ruiz J, Barat JM et al (2010) Influence of pre-cure freezing of Iberian ham on proteolytic changes throughout the ripening process. Meat Sci 85(1):121–126. https://doi.org/10.1016/j.meatsci.2009.12.015
Pérez-Rodríguez J, Téllez-Jurado A, Villa-Tanaca L et al (2022) Intracellular aminopeptidase activity determination from the fungus Sporisorium reilianum: purification and biochemical characterization of psrAPEi enzyme. Curr Microbiol 79(3):90. https://doi.org/10.1007/s00284-022-02787-8
Punia S, Sandhu KS, Grasso S et al (2020) Aspergillus oryzae fermented rice bran: a byproduct with enhanced bioactive compounds and antioxidant potential. Foods 10(1):70. https://doi.org/10.3390/foods10010070
Qin Q, Tang C, Wu J et al (2021) A dual-functional aminopeptidase from Streptomyces canus T20 and its application in the preparation of small rice peptides. Int J Biol Macromol 167:214–222. https://doi.org/10.1016/j.ijbiomac.2020.11.175
Rahulan R, Dhar KS, Nampoothiri KM et al (2012) Characterization of leucine amino peptidase from Streptomyces gedanensis and its applications for protein hydrolysis. Process Biochem 47(2):234–242. https://doi.org/10.1016/j.procbio.2011.10.038
Razzaq A, Shamsi S, Ali A et al (2019) Microbial proteases applications. Front Bioeng Biotech 7:110. https://doi.org/10.3389/fbioe.2019.00110
Santos K, Medrano FJ (2007) Expression, purification, and characterization of an aminopeptidase (Xac2987) with broad specificity from Xanthomonas axonopodis pv. citri. Protein Expres Purif 52(1):117–122. https://doi.org/10.1016/j.pep.2006.10.001
Soeryapranata E, Powers JR, Unlu G (2007) Cloning and characterization of debittering peptidases, PepE, PepO, PepO2, PepO3, and PepN, of Lactobacillus helveticus WSU19. Int Dairy J 17(9):1096–1106. https://doi.org/10.1016/j.idairyj.2007.02.002
Solanki P, Putatunda C, Kumar A et al (2021) Microbial proteases: ubiquitous enzymes with innumerable uses. 3 Biotech 11(10):428. https://doi.org/10.1007/s13205-021-02928-z
Song P, Feng W (2021) Functional expression and characterization of a novel aminopeptidase B from Aspergillus niger in Pichia pastoris. 3 Biotech 11(8):366. https://doi.org/10.1007/s13205-021-02915-4
Song P, Cheng L, Tian K et al (2020) A novel aminopeptidase with potential debittering properties in casein and soybean protein hydrolysates. Food Sci Biotechnol 29:1491–1499. https://doi.org/10.1007/s10068-020-00813-8
Souza TSP, de Andrade CJ, Koblitz MGB et al (2022) Microbial oeptidase in food processing: current state of the art and future trends. Catal Lett. https://doi.org/10.1007/s10562-022-03965-w
Story SV, Shah C, Jenney FE et al (2005) Characterization of a novel zinc-containing, lysine-specific aminopeptidase from the hyperthermophilic archaeon Pyrococcus furiosus. J Bacteriol 187(6):2077–2083. https://doi.org/10.1128/JB.187.6.2077-2083.2005
Stressler T, Pfahler N, Merz M et al (2016) A fusion protein consisting of the exopeptidases PepN and PepX-production, characterization, and application. Appl Microbiol Biotechnol 100(17):7499–7515. https://doi.org/10.1007/s00253-016-7478-8
Stressler T, Tanzer C, Ewert J et al (2017) Simple purification method for a recombinantly expressed native His-tag-free aminopeptidase A from Lactobacillus delbrueckii. Protein Expres Purif 131:7–15. https://doi.org/10.1016/j.pep.2016.10.010
Tang XL, Lu XF, Wu ZM et al (2018) Biocatalytic production of (S)-2-aminobutanamide by a novel d-aminopeptidase from Brucella sp. with high activity and enantioselectivity. J Biotechnol 266:20–26. https://doi.org/10.1016/j.jbiotec.2017.12.003
Tong X, Lian Z, Miao L et al (2020) An innovative two-step enzyme-assisted aqueous extraction for the production of reduced bitterness soybean protein hydrolysates with high nutritional value. LWT-Food Sci Technol 134:110151. https://doi.org/10.1016/j.lwt.2020.110151
Udenigwe CC (2014) Bioinformatics approaches, prospects and challenges of food bioactive peptide research. Trends Food Sci Tech 36(2):137–143. https://doi.org/10.1016/j.tifs.2014.02.004
Vanunu M, Schall P, Reingewertz TH et al (2019) MapB protein is the essential methionine aminopeptidase in Mycobacterium tuberculosis. Cells 8:393. https://doi.org/10.3390/cells8050393
Vargas F, Wangesteen R, Rodríguez-Gómez I et al (2020) Aminopeptidases in cardiovascular and renal function. Role as predictive renal injury biomarkers. Int J Mol Sci 21(16):5615. https://doi.org/10.3390/ijms21165615
Wan K, Uraji M, Tokai S et al (2019) Enzymatic degradation of allergen peptides from bovine casein by a combination of Streptomyces aminopeptidases. Appl Biochem Biotechnol 187(2):570–582. https://doi.org/10.1007/s12010-018-2839-7
Wan K, Uraji M, Yang L et al (2020) Novel activity of Streptomyces aminopeptidase P. Bioresour Bioprocess 7:20. https://doi.org/10.1186/s40643-020-00309-7
Wang Z, Liu J, Xu L et al (2013) Preparation of optically active alkoxy-serines from amino-amide racemate catalyzed by Escherichia coli cells with peptidase B activity. Chem Res Chinese U 29(1):95–98. https://doi.org/10.1007/s40242-012-2249-2
Woessner JF (2013) Chapter 386 - Clostridial Aminopeptidase. In: Handbook of Proteolytic Enzymes (Third Edition), pp. 1699–1701. https://doi.org/10.1016/B978-0-12-382219-2.00386-0
Xi H, Tian Y, Zhou N et al (2015) Characterization of an N-glycosylated Bacillus subtilis leucine aminopeptidase expressed in Pichia pastoris. J Basic Microb 55(2):236–246. https://doi.org/10.1002/jobm.201400368
Xu JH, Jiang Z, Solania A et al (2018) A commensal dipeptidyl aminopeptidase with specificity for N-terminal glycine degrades human-produced antimicrobial peptides in vitro. ACS Chem Biol 13(9):2513–2521. https://doi.org/10.1021/acschembio.8b00420
Yang H, Zhu Q, Zhou N et al (2016) Optimized expression of prolyl aminopeptidase in Pichia pastoris and its characteristics after glycosylation. World J Microbiol Biotechnol 32(11):176. https://doi.org/10.1007/s11274-016-2135-z
Yoon J, Sekhon SS, Kim YH et al (2016) Enhanced lysosomal activity by overexpressed aminopeptidase Y in Saccharomyces cerevisiae. Mol Cell Biochem 417(1–2):181–189. https://doi.org/10.1007/s11010-016-2728-8
Zdunek-Zastocka E, Grabowska A, Branicki T et al (2017) Biochemical characterization of the triticale TsPAP1, a new type of plant prolyl aminopeptidase, and its impact on proline content and flowering time in transgenic Arabidopsis plants. Plant Physiol Bioch 116:18–26. https://doi.org/10.1016/j.plaphy.2017.04.026
Zhang JH, Jin GF, Wang JM et al (2011) Effect of intensifying high-temperature ripening on lipolysis and lipid oxidation of Jinhua ham. LWT-Food Sci Technol 44(2):473–479. https://doi.org/10.1016/j.lwt.2010.07.007
Zhang L, Cai QF, Wu GP et al (2013) Arginine aminopeptidase from white shrimp (Litopenaeus vannamei) muscle: purification and characterization. Eur Food Res Technol 236(5):759–769. https://doi.org/10.1007/s00217-013-1941-x
Zhao P, Zhang M, Wan X et al (2022) A novel Co2+-dependent leucyl aminopeptidase Amp0279 originated from Lysinibacillus sphaericus and heterologous expression using different expression systems. Appl Microbiol Biotechnol 106:1139–1149. https://doi.org/10.1007/s00253-022-11767-8
Funding
This work was supported by the National Natural Science Foundation of China (32170008, 32211530564), Hubei Natural Science Foundation (2021CFB201), the Key R&D projects in Hubei Province (2021BCA113), and Hubei Province Innovation Major Program (2018ABA093).
Author information
Authors and Affiliations
Contributions
YW and XH wrote the main manuscript text. PZ, YZ and HX prepared Table 1. All authors reviewed the manuscript.
Corresponding authors
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Wang, Y., Zhao, P., Zhou, Y. et al. From bitter to delicious: properties and uses of microbial aminopeptidases. World J Microbiol Biotechnol 39, 72 (2023). https://doi.org/10.1007/s11274-022-03501-3
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s11274-022-03501-3