Characteristics of the Proteolytic Enzymes Produced by Lactic Acid Bacteria

Over the past several decades, we have observed a very rapid development in the biotechnological use of lactic acid bacteria (LAB) in various branches of the food industry. All such areas of activity of these bacteria are very important and promise enormous economic and industrial successes. LAB are a numerous group of microorganisms that have the ability to ferment sugars into lactic acid and to produce proteolytic enzymes. LAB proteolytic enzymes play an important role in supplying cells with the nitrogen compounds necessary for their growth. Their nutritional requirements in this regard are very high. Lactic acid bacteria require many free amino acids to grow. The available amount of such compounds in the natural environment is usually small, hence the main function of these enzymes is the hydrolysis of proteins to components absorbed by bacterial cells. Enzymes are synthesized inside bacterial cells and are mostly secreted outside the cell. This type of proteinase remains linked to the cell wall structure by covalent bonds. Thanks to advances in enzymology, it is possible to obtain and design new enzymes and their preparations that can be widely used in various biotechnological processes. This article characterizes the proteolytic activity, describes LAB nitrogen metabolism and details the characteristics of the peptide transport system. Potential applications of proteolytic enzymes in many industries are also presented, including the food industry.

[1]  T. Tamilselvan,et al.  Role of enzymes for improvement in gluten-free foxtail millet bread: It’s effect on quality, textural, rheological and pasting properties , 2021 .

[2]  D. Agyei,et al.  Cell-envelope proteinases from lactic acid bacteria: Biochemical features and biotechnological applications. , 2020, Comprehensive reviews in food science and food safety.

[3]  L. Nouri,et al.  Evaluating the effects of lactic acid bacteria and olive leaf extract on the quality of gluten-free bread , 2020 .

[4]  C. Cambillau,et al.  Wine Phenolic Compounds Differently Affect the Host-Killing Activity of Two Lytic Bacteriophages Infecting the Lactic Acid Bacterium Oenococcus oeni , 2020, Viruses.

[5]  G. Nevárez-Moorillón,et al.  Selection of Lactic Acid Bacteria Isolated from Fresh Fruits and Vegetables Based on Their Antimicrobial and Enzymatic Activities , 2020, Foods.

[6]  D. Witrowa‐Rajchert,et al.  The influence of Lactobacillus bacteria type and kind of carrier on the properties of spray‐dried microencapsules of fermented beetroot powders , 2020, International Journal of Food Science & Technology.

[7]  P. Puligundla,et al.  Recent developments in high gravity beer-brewing , 2020 .

[8]  B. Martínez,et al.  Polyphasic Characterisation of Non-Starter Lactic Acid Bacteria from Algerian Raw Camel’s Milk and Their Technological Aptitudes , 2020, Food technology and biotechnology.

[9]  J. Zufía,et al.  Brewers’ Spent Yeast and Grain Protein Hydrolysates as Second-Generation Feedstuff for Aquaculture Feed , 2020, Waste and Biomass Valorization.

[10]  G. Pérez-Martínez,et al.  Differences in the expression of cell envelope proteinases (CEP) in two Lactobacillus paracasei probiotics strains. , 2020, FEMS microbiology letters.

[11]  B. Teusink,et al.  Enhancement of amino acid production and secretion by Lactococcus lactis using a droplet-based biosensing and selection system , 2020, Metabolic engineering communications.

[12]  A. Sussulini,et al.  Proteolytic enzymes positively modulated the physicochemical and antioxidant properties of spent yeast protein hydrolysates , 2020 .

[13]  Jing Qi,et al.  Effects of lactic acid fermentation-based biotransformation on phenolic profiles, antioxidant capacity and flavor volatiles of apple juice , 2020 .

[14]  Yingying Hu,et al.  Production, purification and biochemical characterization of the microbial protease produced by Lactobacillus fermentum R6 isolated from Harbin dry sausages , 2020 .

[15]  S. Walker,et al.  Uncovering the activities, biological roles, and regulation of bacterial cell wall hydrolases and tailoring enzymes , 2020, The Journal of Biological Chemistry.

[16]  Braddock A Sandoval,et al.  Emerging strategies for expanding the toolbox of enzymes in biocatalysis. , 2020, Current opinion in chemical biology.

[17]  G. Galaverna,et al.  Application of lactic acid fermentation to elderberry juice: Changes in acidic and glucidic fractions , 2020 .

[18]  J. Suárez,et al.  Adherence of Lactobacillus salivarius to HeLa Cells Promotes Changes in the Expression of the Genes Involved in Biosynthesis of Their Ligands , 2020, Frontiers in Immunology.

[19]  W. Białas,et al.  Integrated approach for obtaining bioactive peptides from whey proteins hydrolysed using a new proteolytic lactic acid bacteria. , 2019, Food chemistry.

[20]  D. Tagliazucchi,et al.  Bioprospecting for Bioactive Peptide Production by Lactic Acid Bacteria Isolated from Fermented Dairy Food , 2019, Fermentation.

[21]  Erna Normaya Abdullah,et al.  Optimization of a protease extraction using a statistical approach for the production of an alternative meat tenderizer fromSpondias cytherearoots , 2019, Journal of Food Processing and Preservation.

[22]  M. Ashokkumar,et al.  Effects of high pressure, microwave and ultrasound processing on proteins and enzyme activity in dairy systems — A review , 2019, Innovative Food Science & Emerging Technologies.

[23]  C. Peláez,et al.  Expression of amino acid converting enzymes and production of volatile compounds by Lactococcus lactis IFPL953 , 2019, International Dairy Journal.

[24]  Raman Kumar,et al.  Next generation sequencing, biochemical characterization, metabolic pathway analysis of novel probiotic Pediococcus acidilactici NCDC 252 and it’s evolutionary relationship with other lactic acid bacteria , 2019, Molecular Biology Reports.

[25]  C. Hidayat,et al.  Isolation, Screening, and Identification of Proteolytic Lactic Acid Bacteria from Indigenous Chao Product , 2019, Journal of Aquatic Food Product Technology.

[26]  B. Kong,et al.  Purification and biochemical characteristics of the microbial extracellular protease from Lactobacillus curvatus isolated from Harbin dry sausages. , 2019, International journal of biological macromolecules.

[27]  N. O'Brien,et al.  Characterisation of the in vitro bioactive properties of alkaline and enzyme extracted brewers' spent grain protein hydrolysates. , 2019, Food research international.

[28]  Q. Ali,et al.  Microbial Proteases Applications , 2019, Front. Bioeng. Biotechnol..

[29]  Cristóbal N. Aguilar,et al.  Production of Bioactive Peptides from Lactic Acid Bacteria: A Sustainable Approach for Healthier Foods. , 2019, Comprehensive reviews in food science and food safety.

[30]  N. Nordin,et al.  Microbial hydrolytic enzymes: In silico studies between polar and tropical regions , 2019, Polar Science.

[31]  P. Prabhasankar,et al.  Targeted degradation of gluten proteins in wheat flour by prolyl endoprotease and its utilization in low immunogenic pasta for gluten sensitivity population , 2019, Journal of Cereal Science.

[32]  L. Nissen,et al.  Metabolomic approach to study the impact of flour type and fermentation process on volatile profile of bakery products. , 2019, Food research international.

[33]  B. Ramakrishnan,et al.  A systematic reconsideration on proteases. , 2019, International journal of biological macromolecules.

[34]  I. García-Cano,et al.  Lactic acid bacteria isolated from dairy products as potential producers of lipolytic, proteolytic and antibacterial proteins , 2019, Applied Microbiology and Biotechnology.

[35]  Ashwani Kumar,et al.  A Review on Microbial Alkaline Protease: An Essential Tool for Various Industrial Approaches , 2019, Industrial Biotechnology.

[36]  Rosfarizan Mohamad,et al.  Extracellular Proteolytic Activity and Amino Acid Production by Lactic Acid Bacteria Isolated from Malaysian Foods , 2019, International journal of molecular sciences.

[37]  Rosfarizan Mohamad,et al.  Comparative studies of versatile extracellular proteolytic activities of lactic acid bacteria and their potential for extracellular amino acid productions as feed supplements , 2019, Journal of Animal Science and Biotechnology.

[38]  W. Białas,et al.  Identification and partial characterization of proteolytic activity of Enterococcus faecalis relevant to their application in dairy industry. , 2019, Acta biochimica Polonica.

[39]  D. Kołożyn-Krajewska,et al.  Effect of pullulan on physicochemical, microbiological, and sensory quality of yogurts. , 2019, Current pharmaceutical biotechnology.

[40]  Pradeep K. Singh,et al.  Enzymes in the Meat Industry , 2019, Enzymes in Food Biotechnology.

[41]  E. Abada Application of Microbial Enzymes in the Dairy Industry , 2019, Enzymes in Food Biotechnology.

[42]  D. Habermann,et al.  Soluble Lactobacillus delbrueckii subsp. bulgaricus 92059 PrtB proteinase derivatives for production of bioactive peptide hydrolysates from casein , 2019, Applied Microbiology and Biotechnology.

[43]  D. Ow,et al.  Brewing with malted barley or raw barley: what makes the difference in the processes? , 2018, Applied Microbiology and Biotechnology.

[44]  L. Fischer,et al.  Improving the colloidal and sensory properties of a caseinate hydrolysate using particular exopeptidases. , 2018, Food & function.

[45]  D. Tagliazucchi,et al.  Peptidomic study of casein proteolysis in bovine milk by Lactobacillus casei PRA205 and Lactobacillus rhamnosus PRA331 , 2018, International Dairy Journal.

[46]  D. Kołożyn-Krajewska,et al.  Food-Origin Lactic Acid Bacteria May Exhibit Probiotic Properties: Review , 2018, BioMed research international.

[47]  F. Toldrá,et al.  Bioactive peptides as natural antioxidants in food products – A review , 2018, Trends in Food Science & Technology.

[48]  Y. Hao,et al.  Global Transcriptomic Analysis and Function Identification of Malolactic Enzyme Pathway of Lactobacillus paracasei L9 in Response to Bile Stress , 2018, Front. Microbiol..

[49]  R. Hutkins,et al.  Probiotics for human use , 2018, Nutrition Bulletin.

[50]  H. Rabesona,et al.  Brazilian artisanal ripened cheeses as sources of proteolytic lactic acid bacteria capable of reducing cow milk allergy , 2018, Journal of applied microbiology.

[51]  E. Daliri,et al.  Antihypertensive peptides from whey proteins fermented by lactic acid bacteria , 2018, Food Science and Biotechnology.

[52]  F. Assefa,et al.  The Role of Microbial Aspartic Protease Enzyme in Food and Beverage Industries , 2018, Journal of Food Quality.

[53]  R. Sidari,et al.  Sourdoughs as a source of lactic acid bacteria and yeasts with technological characteristics useful for improved bakery products , 2018, European Food Research and Technology.

[54]  P. B. Devi,et al.  Recent developments on encapsulation of lactic acid bacteria as potential starter culture in fermented foods – a review , 2018 .

[55]  H. Sato,et al.  Microbial proteases: Production and application in obtaining protein hydrolysates. , 2018, Food research international.

[56]  Taketo Wakai,et al.  Genome-wide motif predictions of BCARR-box in the amino-acid repressed genes of Lactobacillus helveticus CM4 , 2017, BMC Microbiology.

[57]  Arbakariya B. Ariff,et al.  Extractive Fermentation of Lactic Acid in Lactic Acid Bacteria Cultivation: A Review , 2017, Front. Microbiol..

[58]  Bo Zhang,et al.  Characterization of a lactose-responsive promoter of ATP-binding cassette (ABC) transporter gene from Lactobacillus acidophilus 05–172 , 2017, FEMS microbiology letters.

[59]  F. Mozzi,et al.  YebC, a putative transcriptional factor involved in the regulation of the proteolytic system of Lactobacillus , 2017, Scientific Reports.

[60]  C. Montanari,et al.  New bread formulation with improved rheological properties and longer shelf-life by the combined use of transglutaminase and sourdough , 2017 .

[61]  J. Qiao,et al.  Improving nitrogen source utilization from defatted soybean meal for nisin production by enhancing proteolytic function of Lactococcus lactis F44 , 2017, Scientific Reports.

[62]  Qinglong Wu,et al.  High γ-aminobutyric acid production from lactic acid bacteria: Emphasis on Lactobacillus brevis as a functional dairy starter , 2017, Critical reviews in food science and nutrition.

[63]  M. Cocaign-Bousquet,et al.  From Genome to Phenotype: An Integrative Approach to Evaluate the Biodiversity of Lactococcus lactis , 2017, Microorganisms.

[64]  Long Liu,et al.  Microbial response to environmental stresses: from fundamental mechanisms to practical applications , 2017, Applied Microbiology and Biotechnology.

[65]  Guoyao Wu,et al.  Protein hydrolysates in animal nutrition: Industrial production, bioactive peptides, and functional significance , 2017, Journal of Animal Science and Biotechnology.

[66]  M. Kieliszek,et al.  The Effect of Pullulan on the Growth and Acidifying Activity of Selected Stool Microflora of Human. , 2017, Current pharmaceutical biotechnology.

[67]  F. Mozzi,et al.  Lactic Acid Bacteria as Cell Factories for the Generation of Bioactive Peptides. , 2017, Protein and peptide letters.

[68]  R. R. da Silva,et al.  Bacterial and Fungal Proteolytic Enzymes: Production, Catalysis and Potential Applications , 2017, Applied Biochemistry and Biotechnology.

[69]  M. Sohaib,et al.  Plant and bacterial proteases: A key towards improving meat tenderization, a mini review , 2016 .

[70]  R. D. de Vries,et al.  Protease and lipase activities of fungal and bacterial strains derived from an artisanal raw ewe's milk cheese. , 2016, International journal of food microbiology.

[71]  A. Schieber,et al.  Formation of taste-active amino acids, amino acid derivatives and peptides in food fermentations - A review. , 2016, Food research international.

[72]  M. Hollenberg,et al.  Proteinases, Their Extracellular Targets, and Inflammatory Signaling , 2016, Pharmacological Reviews.

[73]  T. Guo,et al.  Characterization of a New Cell Envelope Proteinase PrtP from Lactobacillus rhamnosus CGMCC11055. , 2016, Journal of agricultural and food chemistry.

[74]  W. Galia,et al.  Acquisition of PrtS in Streptococcus thermophilus is not enough in certain strains to achieve rapid milk acidification , 2016 .

[75]  G. Savoy de Giori,et al.  Characterization of the mature cell surface proteinase of Lactobacillus delbrueckii subsp. lactis CRL 581 , 2014, Applied Microbiology and Biotechnology.

[76]  L. Miclo,et al.  Use of a free form of the Streptococcus thermophilus cell envelope protease PrtS as a tool to produce bioactive peptides , 2014 .

[77]  A. Zambrowicz,et al.  Biological and functional properties of proteolytic enzyme-modified egg protein by-products , 2013, Food science & nutrition.

[78]  M. Griffiths,et al.  Lactobacillus helveticus: the proteolytic system , 2012, Front. Microbiol..

[79]  M. Kieliszek,et al.  PURIFICATION AND CHARACTERIZATION OF A PROTEINASE FROM THE PROBIOTIC Lactobacillus rhamnosus OXY , 2012, Preparative biochemistry & biotechnology.

[80]  B. Kuster,et al.  Lactocepin secreted by Lactobacillus exerts anti-inflammatory effects by selectively degrading proinflammatory chemokines. , 2012, Cell host & microbe.

[81]  L. Miclo,et al.  Variability of hydrolysis of β-, αs1-, and αs2-caseins by 10 strains of Streptococcus thermophilus and resulting bioactive peptides. , 2012, Journal of agricultural and food chemistry.

[82]  T. Haertlé,et al.  Comparative analysis of β-casein proteolysis by PrtP proteinase from Lactobacillus paracasei subsp. paracasei BGHN14, PrtR proteinase from Lactobacillus rhamnosus BGT10 and PrtH proteinase from Lactobacillus helveticus BGRA43 , 2011 .

[83]  Y. Ardö,et al.  Variation in aminopeptidase and aminotransferase activities of six cheese related Lactobacillus helveticus strains , 2010 .

[84]  M. Kojić,et al.  The presence of prtP proteinase gene in natural isolate Lactobacillus plantarum BGSJ3–18 , 2010, Letters in applied microbiology.

[85]  R. Mahajan,et al.  Biological aspects of proteolytic enzymes: A Review , 2010 .

[86]  S. Lortal,et al.  prtH2, Not prtH, Is the Ubiquitous Cell Wall Proteinase Gene in Lactobacillushelveticus , 2009, Applied and Environmental Microbiology.

[87]  T. Zotta,et al.  Enzymatic activities of lactic acid bacteria isolated from Cornetto di Matera sourdoughs. , 2007, International journal of food microbiology.

[88]  H. Ingmer,et al.  Proteolytic systems of lactic acid bacteria , 2006, Applied Microbiology and Biotechnology.

[89]  G. Cichosz,et al.  Aktywnosc peptydazowa wybranych szczepow Lactobacillus , 2006 .

[90]  G. Jovanovic,et al.  Analysis of the presence of prtR proteinase gene in natural isolates of Lactobacillus rhamnosus. , 2006, Folia microbiologica.

[91]  A. Trubuil,et al.  Proteomic Signature of Lactococcus lactis NCDO763 Cultivated in Milk , 2005, Applied and Environmental Microbiology.

[92]  R. Vogel,et al.  Functional Characterization of the Proteolytic System of Lactobacillus sanfranciscensis DSM 20451T during Growth in Sourdough , 2005, Applied and Environmental Microbiology.

[93]  S. Oliver,et al.  Glutamic protease distribution is limited to filamentous fungi. , 2004, FEMS microbiology letters.

[94]  P. Renault,et al.  Intracellular effectors regulating the activity of the Lactococcus lactis CodY pleiotropic transcription regulator , 2004, Molecular microbiology.

[95]  D. le Bars,et al.  Proteome Analyses of Heme-Dependent Respiration in Lactococcus lactis: Involvement of the Proteolytic System , 2004, Journal of bacteriology.

[96]  R. Siezen Multi-domain, cell-envelope proteinases of lactic acid bacteria , 1999, Antonie van Leeuwenhoek.

[97]  G. Krasnowska Próba wykorzystania enzymów pochodzenia mikrobiologicznego do degradacji surowców zwierzęcych bogatych w tkankę łączną , 2004 .

[98]  M. Kleerebezem,et al.  Identification and Genetic Characterization of a Novel Proteinase, PrtR, from the Human Isolate Lactobacillus rhamnosus BGT10 , 2003, Applied and Environmental Microbiology.

[99]  K. Jordan,et al.  Growth phase and growth medium effects on the peptidase activities of Lactobacillus helveticus , 2003 .

[100]  P. Renault,et al.  Transcriptional Pattern of Genes Coding for the Proteolytic System of Lactococcus lactis and Evidence for Coordinated Regulation of Key Enzymes by Peptide Supply , 2001, Journal of bacteriology.

[101]  P. Renault,et al.  Pleiotropic transcriptional repressor CodY senses the intracellular pool of branched‐chain amino acids in Lactococcus lactis , 2001, Molecular microbiology.

[102]  J. Björkroth,et al.  Taxonomy and important features of probiotic microorganisms in food and nutrition. , 2001, The American journal of clinical nutrition.

[103]  A. Galinier,et al.  Autoregulation of the biosynthesis of the CcpA-like protein, PepR1, in Lactobacillus delbrueckii subsp bulgaricus. , 2001, Journal of molecular microbiology and biotechnology.

[104]  R. Raya,et al.  Nutritional Requirements and Nitrogen-Dependent Regulation of Proteinase Activity of Lactobacillus helveticus CRL 1062 , 2000, Applied and Environmental Microbiology.

[105]  R. Tampé,et al.  Combinatorial peptide libraries reveal the ligand-binding mechanism of the oligopeptide receptor OppA of Lactococcus lactis. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[106]  V. Monnet,et al.  Streptococcus thermophilus Cell Wall-Anchored Proteinase: Release, Purification, and Biochemical and Genetic Characterization , 2000, Applied and Environmental Microbiology.

[107]  D. Aubel,et al.  Characterization of a prolidase from Lactobacillus delbrueckii subsp. bulgaricus CNRZ 397 with an unusual regulation of biosynthesis. , 1999, Microbiology.

[108]  Edmund R. S. Kunji,et al.  Kinetics and specificity of peptide uptake by the oligopeptide transport system of Lactococcus lactis. , 1998, Biochemistry.

[109]  A. Hagting,et al.  Cloning and functional expression in Escherichia coli of the gene encoding the di- and tripeptide transport protein of Lactobacillus helveticus , 1997, Applied and environmental microbiology.

[110]  J. R. Reid,et al.  Involvement of enzyme-substrate charge interactions in the caseinolytic specificity of lactococcal cell envelope-associated proteinases , 1995, Applied and environmental microbiology.

[111]  A. Hagting,et al.  Specificity of peptide transport systems in Lactococcus lactis: evidence for a third system which transports hydrophobic di- and tripeptides , 1995, Journal of bacteriology.

[112]  A. Bruins,et al.  The extracellular PI-type proteinase of Lactococcus lactis hydrolyzes beta-casein into more than one hundred different oligopeptides , 1995, Journal of bacteriology.

[113]  A. Hagting,et al.  Transport of -Casein-derived Peptides by the Oligopeptide Transport System Is a Crucial Step in the Proteolytic Pathway of Lactococcus lactis(*) , 1995, The Journal of Biological Chemistry.

[114]  G. Venemâ,et al.  Genetic manipulation of the peptidolytic system in lactic acid bacteria , 1995 .

[115]  O. Schneewind,et al.  Proteolytic cleavage and cell wall anchoring at the LPXTG motif of surface proteins in Gram‐positive bacteria , 1994, Molecular microbiology.

[116]  T. Coolbear,et al.  The physiology and biochemistry of the proteolytic system in lactic acid bacteria. , 1993, FEMS microbiology reviews.

[117]  J. Kok Genetics of Proteolytic Enzymes of Lactococci and Their Role in Cheese Flavor Development , 1993 .

[118]  A. Holck,et al.  Cloning, sequencing and expression of the gene encoding the cell-envelope-associated proteinase from Lactobacillus paracasei subsp. paracasei NCDO 151. , 1992, Journal of general microbiology.

[119]  N. Haard,et al.  A Review of Proteotlytic Enzymes from Marine Organisms and Their Application in the Food Industry , 1992 .

[120]  W. D. de Vos,et al.  Proteinase overproduction in Lactococcus lactis strains: regulation and effect on growth and acidification in milk , 1992, Applied and environmental microbiology.

[121]  G. Venema,et al.  Lactococcal proteinase maturation protein PrtM is a lipoprotein , 1991, Journal of bacteriology.

[122]  W. D. de Vos,et al.  A maturation protein is essential for production of active forms of Lactococcus lactis SK11 serine proteinase located in or secreted from the cell envelope , 1989, Journal of bacteriology.

[123]  G. Venemâ,et al.  Identification of a gene required for maturation of an extracellular lactococcal serine proteinase , 1989, Journal of bacteriology.