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Cited by 209Enzyme Production and CharacterizationProteins in Food SystemsPineapple and bromelain studies

Plant proteases as milk-clotting enzymes in cheesemaking: a review

Open access
Shah, Manzoor Ahmad, Mir, Shabir Ahmad, Paray, Mohd Amir·2014·Dairy Science & Technology
DOI 10.1007/s13594-013-0144-3
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Abstract

Plant proteases have been used as milk coagulants in cheesemaking for centuries either as crude extracts or in purified form.

Integrity signals

Registry

Crossref · 10.1007/s13594-013-0144-3

Cites

78 of 78 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

6 of 100 cited sources checked · 4 supported · 0 flagged · accuracy not yet measured

  • Premisecitation

    Aspartic proteases with milk-clotting activity have been reported in artichoke, milk thistle, Onopordum turcicum, rice kernels, and Centaurea calcitrapa.

    “Aspartic proteases with milk-clotting activity have been reported in artichoke (Cynara scolymus L.) (Llorente et al. 1997); milk thistle (Silybum marianum L. Gaertn.)(Vairo-Cavalli et al. 2005); Onopordum turcicum (Tamer 1993); rice kernels (Asakura et al. 1997); Centaurea calcitrapa (Domingos et al. 2000).”
    Quote in the paperNot a substantial claim1 of 3 sources checked
    • cites Llorente et al. 1997
    • cites Vairo-Cavalli et al. 2005 source
    • cites Tamer 1993 source
  • Premisecitation

    Precipitation with ammonium sulfate is an effective way to produce substantial amounts of active proteases from the flowers of C. cardunculus.

    “Precipitation with ammonium sulfate is an effective way to produce substantial amounts of active proteases from the flowers of C. cardunculus (Barros et al. 2001).”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites Barros et al. 2001 source
  • Premisecitation

    Aspartic proteases from C. cardunculus flowers share specificity and kinetic parameters with chymosin and pepsin.

    “These flowers contain aspartic proteases, previously described, which have been shown to share specificity and kinetic parameters with chymosin and pepsin (Verissimo et al. 1995(Verissimo et al. , 1996)).”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites Verissimo et al. 1995 source
    • cites Verissimo et al. 1996 source
  • Premisecitation

    Extracts of Cynara spp. have been used in making Portuguese Serra and Serpa cheeses, Spanish Los Pedroches, La Serena, Torta del Casar, Los Ibores, and Flor de Guía cheeses.

    “The extracts of Cynara spp.have been used in the making of Portuguese Serra and Serpa cheeses (Macedo et al. 1993) and Spanish Los Pedroches, La Serena (Roa et al. 1999) and Torta del Casar cheeses (from ewes' milk) as well as Los Ibores cheese (from goats' milk) and Flor de Guía cheese (from a mixture of ewes' and cows' milk) (Fernández-Salguero et al. 1991;Fernández-Salguero 1999;Sanjuán et al.”
    Quote in the paperNot a substantial claim1 of 3 sources checked
    • cites Macedo et al. 1993 source

      The Introduction (page not provided) says cardoon protease is traditionally used in Portugal to make Serra cheese. It does not mention Serpa cheese. The Spanish cheeses in the citing sentence are attributed to other citations, not to Macedo et al. (LLM, conf=0.88)

      Experimental — accuracy not yet measured
    • cites Roa et al. 1999 source
    • cites Fernández-Salguero et al. 1991 source
  • Findingnone given

    Almost all enzymes used as milk coagulants belong to aspartic proteases.

    “Almost all the enzymes used as milk coagulants belong to aspartic proteases, but enzymes from other groups such as cysteine and serine proteases have also been reported and possess the ability to clot milk under proper conditions.”
    • Also stated in: § 1 Introduction · plain
      “Almost all the enzymes used as milk coagulants belong to aspartic proteases, but enzymes from other groups such as cysteine and serine proteases have also been used.”
    Quote in the paperNot a substantial claim
  • Classified as probably not a substantial claim. 25
  • May combine two claims. 28

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Cardoon flowers are traditionally used in the Mediterranean region for cheesemaking.

    “Cardoon (Cynara cardunculus) flowers are traditionally used in the Mediterranean region for cheesemaking (Barros et al. 2003).”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites Barros et al. 2003 source

  • Classified as probably not a substantial claim.

    Premisecitation

    When a potential rennet substitute is studied, it is important to evaluate degradation patterns of caseins because of their effects on yield, consistency and flavor of final cheese.

    “When a potential rennet substitute is studied, it is particularly important to evaluate adequately the degradation patterns of the caseins because of their effects on yield, consistency and flavor of the final cheese (Fox 1989).”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites Fox 1989 source

      The Primary Phase of Rennet Action section says rennet substitutes may differ in the extent or specificity of casein hydrolysis, and the Significance of Proteolysis section links protein breakdown to cheese flavor and texture. The Discussion of premanufacture proteolysis links some proteinase activity to reduced cheese yield, but the paper does not specifically establish a yield effect for rennet substitutes or explicitly recommend evaluating their casein-degradation patterns. (LLM, conf=0.83)

      Experimental — accuracy not yet measured
    • Rests on an assumption: A good milk-clotting enzyme is characterized by high specific caseinolytic activity and low general proteolytic activity.

  • May combine two claims.

    Findingcitation

    Enzymes from cysteine and serine proteases have been reported as milk coagulants and can clot milk under proper conditions.

    “but enzymes from other groups such as cysteine and serine proteases have also been reported and possess the ability to clot milk under proper conditions.”
    • Also stated in: § 1 Introduction · plain
      “but the plant proteases used as milk coagulants have been reported only from first three types (Table 1) and none from metalloproteases.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Table 1

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • May combine two claims.

    Findingown data

    Plant proteases used as milk coagulants have been reported only from aspartate, serine, and cysteine proteases, and none from metalloproteases.

    “but the plant proteases used as milk coagulants have been reported only from first three types (Table 1) and none from metalloproteases.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Table 1

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • May combine two claims.

    Premisecitation

    The excessive proteolytic nature of most plant coagulants limits their use in cheese manufacturing, causing lower cheese yield and flavor and texture defects.

    “The excessive proteolytic nature of most plant coagulants has limited their use in cheese manufacturing due to lower yields of cheese, bitter flavors and texture defects.”
    • Also stated in: § 1 Introduction · plain
      “However, the excessive proteolytic nature of most vegetable coagulants has limited their use in cheese manufacturing due to lower cheese yield and defects in flavor and texture (Lo Piero et al. 2002).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Lo Piero et al. 2002 source

  • May combine two claims.

    Premisecitation

    Cucumisin from Cucumis melo and lettucine from Lactuca sativa were isolated and used as milk coagulants.

    “Cucumisin from Cucumis melo (Uchikoba and Kaneda 1996) and lettucine from Lactuca sativa (Lo Piero et al. 2002) were isolated and used as milk coagulants.”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites Uchikoba and Kaneda 1996 source
    • cites Lo Piero et al. 2002 source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    In West African countries like Nigeria and Benin, extracts from Calotropis procera have been used in traditional cheesemaking.

    “In West African countries like Nigeria and the republic of Benin, extracts from Calotropis procera (Sodom apple) have been used in traditional cheesemaking (Roseiro et al. 2003).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Roseiro et al. 2003 source

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Religiosin (43.3 kDa), Religiosin B (63 kDa), and Religiosin C (80 kDa) were isolated from Ficus religiosa latex.

    “Religiosin (43.3 kDa), Religiosin B (63 kDa) and Religiosin C (80 kDa) were isolated from Ficus religiosa latex by Kumari et al. (2010), Kumari et al. (2012) and Sharma et al. (2012) respectively.”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites Kumari et al. (2010) source
    • cites Kumari et al. (2012) source
    • cites Sharma et al. (2012) source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Cheese production increased by a factor of approximately 3.5 since 1961.

    “The cheese production increased by a factor of approximately 3.5 since 1961 but the rennet supply decreased due to the limited availability of ruminant stomachs (Jacob et al. 2011).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Jacob et al. 2011 source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Plant proteases are found in almost all kinds of plant tissues.

    “These enzymes are found in almost all kinds of plant tissues and can be obtained from their natural source or through in vitro culture to ensure a continuous supply of plant proteases.”
    • Also stated in: § 1 Introduction · hedged
      “According to Tamer and Mavituna (1997), these enzymes are present in almost all kinds of plant tissues and it appears to be a general rule that all proteolytic enzymes have the ability to clot milk under appropriate conditions.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Tamer and Mavituna (1997) source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Three cyprosins with milk-clotting activity from dried flowers of C. cardunculus were isolated, purified, and characterized.

    “Also, three cyprosins with milk-clotting activity, from dried flowers of C. cardunculus, were isolated, purified, and characterized by Heimgartner et al. (1990).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Heimgartner et al. (1990) source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Ginger protease was isolated from ginger rhizomes (Zingiber officinale) having a molecular mass of 36 kDa.

    “Ginger protease was isolated from ginger rhizomes (Zingiber officinale) having a molecular mass of 36 kDa (Hashim et al. 2011).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Hashim et al. 2011 source

  • Classified as probably not a substantial claim.

    Premisecitation

    The search for new potential milk-clotting enzymes from plants continues to meet increasing global demand for diversified and good quality cheese production.

    “The search for new potential milk-clotting enzymes from plants still continues in order to meet the increasing global demand for diversified and good quality cheese production.”
    • Also stated in: § 1 Introduction · plain
      “Therefore, the search for new potential milk-clotting enzymes from plants is in continuous process, so as to make them industrially useful and go with the increasing global demand for diversified and high quality cheese production (Hashim et al. 2011).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Hashim et al. 2011 source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Streblin, a thermostable enzyme having a molecular mass of 63 kDa, was purified from Streblus asper.

    “Streblin, a thermostable enzyme having a molecular mass of 63 kDa was purified from Streblus asper (Tripathi et al. 2011).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Tripathi et al. 2011 source

  • May combine two claims.

    Premisecitation

    Plant coagulants are an alternative to calf rennet because of limited availability and high price of rennet, religious factors, diet, and bans on recombinant calf rennet in some countries.

    “These coagulants are an alternative to the calf rennet due to the limited availability and high price of rennet, religious factors, diet or ban on recombinant calf rennet in some countries.”
    • Also stated in: § 1 Introduction · plain
      “Various factors such as high price of rennet, religious concerns (e.g., Islam and Judaism), diet (vegetarianism) or ban on recombinant calf rennet (in France, Germany and The Netherlands) have encouraged the search for alternative milk-clotting sources (Roseiro et al. 2003).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Roseiro et al. 2003 source

  • May combine two claims.

    Premisecitation

    Cynara cardunculus produces cardosins and cyprosins, aspartic proteases that accumulate in mature flowers (petals and pistils) but not in leaves or seeds.

    “It produces cardosins and cyprosins, aspartic proteases that have been found to accumulate in mature flowers (petals and pistils) but not in leaves or seeds (Cordeiro et al. 1998).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Cordeiro et al. 1998

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • May combine two claims.

    Premisecitation

    Plant proteases can be obtained from their natural source or through in vitro culture to ensure a continuous supply.

    “These enzymes are found in almost all kinds of plant tissues and can be obtained from their natural source or through in vitro culture to ensure a continuous supply of plant proteases.”
    • Also stated in: § 3 Production of plant proteases · plain
      “These enzymes can be obtained from their natural source or through in vitro culture to ensure a continuous supply of plant proteases (Gonzalez-Rabade et al. 2011).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Gonzalez-Rabade et al. 2011 source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Actinidin was isolated from ripe fresh kiwifruits (Actinidia chinensis).

    “Actinidin was isolated from ripe fresh kiwifruits (Actinidia chinensis) (Katsaros et al. 2010).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Katsaros et al. 2010 source

  • May combine two claims.

    Premisecitation

    Aspartic proteases are most active at acidic pH and show preferential specificity for cleavage at peptide bonds between hydrophobic amino acid residues.

    “They are most active at acidic pH and show preferential specificity for cleavage at peptide bonds between hydrophobic amino acid residues responsible for the catalytic activity (Domingos et al. 2000).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Domingos et al. 2000 source

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Dubiumin was purified from the seeds of Solanum dubium, having a molecular mass of 66 kDa.

    “Dubiumin was purified from the seeds of Solanum dubium, having a molecular mass of 66 kDa (Ahmed et al. 2009b).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Ahmed et al. 2009b source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Spain and Portugal have the largest variety and production of cheeses using Cynara sp. as the vegetable coagulant.

    “Spain and Portugal have the largest variety and production of cheeses using Cynara sp. as the vegetable coagulant (Roseiro et al. 2003).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Roseiro et al. 2003 source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    All proteolytic enzymes have the ability to clot milk under appropriate conditions.

    “it appears to be a general rule that all proteolytic enzymes have the ability to clot milk under appropriate conditions.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Tamer and Mavituna (1997) source

  • May combine two claims.

    Interpretationcitation

    Hieronymain might be appropriate for cheesemaking and for production of milk protein hydrolysates.

    “It was concluded that hieronymain might be appropriate for cheesemaking, as well as for the production of milk protein hydrolysates (Bruno et al. 2010).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Bruno et al. 2010 source

  • May combine two claims.

    Premisecitation

    Dubiumin works efficiently under different salt concentrations and at different pH levels.

    “A serine protease named dubiumin, isolated from Solanum dubium seeds, having pI value of 9.3 and optimum pH 11.0 worked efficiently under different salts concentration and at different pH levels.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Ahmed et al. 2009b source

  • May combine two claims.

    Premisecitation

    Cheese made using UF milk and calf rennet was found to be hard and crumbly, with less developed flavors.

    “Cheese made using UF milk and calf rennet was found to be hard and crumbly, with less developed flavors (Renner and Abd El-Salam 1991).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Renner and Abd El-Salam 1991

  • May combine two claims.

    Premisecitation

    Problems due to climate and season conditions, and heterogeneity of the product obtained from plant parts can be solved by in vitro techniques.

    “The problems due to climate and season conditions, and heterogeneity of the product obtained from plant parts can also be solved by in vitro techniques (Gonzalez-Rabade et al. 2011).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Gonzalez-Rabade et al. 2011 source

  • Classified as probably not a substantial claim.

    Premisecitation

    Milk-clotting activity is the capability of the enzyme for specific κ-casein hydrolysis.

    “It is the capability of the enzyme for specific κ-casein hydrolysis (Jacob et al. 2011).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Jacob et al. 2011 source

  • May combine two claims.

    Premisecitation

    While UF may have reduced the extent of milk and cheese proteolysis in the presence of cardoon extract, it did not result in improved sensory qualities unless a significant amount of regular milk was present.

    “Proteolytic, textural, and sensory studies showed that while UF may have reduced the extent of milk and cheese proteolysis in the presence of cardoon extract, it did not result in improved sensory qualities unless a significant amount of regular milk was present (Agboola et al. 2009).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Agboola et al. 2009 source

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Electrophoretic techniques currently used in study of milk proteins (SDS-PAGE, alkaline urea PAGE, acid urea PAGE) usually do not allow clear identification of all casein components, and Tricine SDS-PAGE is recommended.

    “The electrophoretic techniques currently used in the study of milk proteins (SDS-PAGE, alkaline urea PAGE, acid urea PAGE) usually do not allow a clear identification of all casein components and hence an improved electrophoretical technique (Tricine SDS-PAGE) is recommended (Bruno et al. 2010).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Bruno et al. 2010 source
    • Rests on an assumption: One of the most adequate and expeditious methods to monitor proteolytic processes is polyacrylamide gel electrophoresis.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Milk-clotting activity can be measured by different methods such as Soxhlet, Berridge, and international standard method, with units Soxhlet units, Berridge, Rennet units, and International Milk Clotting Units.

    “It can be measured by different methods such as Soxhlet, Berridge, and international standard method and the units used are Soxhlet units, Berridge, or Rennet units and International Milk Clotting Units, respectively (Harboe et al. 2010).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Harboe et al. 2010

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

  • May combine two claims.

    Premisecitation

    Degradation of other casein fractions by hieronymain proceeds slowly enough to guarantee production of a firm curd, with no evidence of extensive hydrolysis.

    “The κ-casein fraction, directly involved in clotting formation, began to be degraded after 10 min of reaction, while the degradation of the other casein fractions proceeds slowly enough as to guarantee the production of a firm curd, with no evidence of extensive hydrolysis.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Bruno et al. 2010 source

  • May combine two claims.

    Premisecitation

    Cheeses prepared with plant coagulants have a characteristic soft texture and a slightly bitter flavor.

    “Cheeses prepared with plants coagulants have a characteristic soft texture and a slightly bitter flavor (Roseiro et al. 2003).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Roseiro et al. 2003 source

  • May combine two claims.

    Premisecitation

    Hieronymain hydrolyzes bovine casein and milk whey proteins.

    “A protease named hieronymain, obtained from unripe fruits of Bromelia hieronymi, was capable of clotting milk and hydrolyzing bovine casein and milk whey proteins.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Bruno et al. 2010 source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Onopordosin has optimum pH 2.5.

    “It is an aspartic protease of optimum pH 2.5 and pI 4.4.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Brutti et al. 2012 source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Dubiumin has a pI of 9.3.

    “A serine protease named dubiumin, isolated from Solanum dubium seeds, having pI value of 9.3 and optimum pH 11.0 worked efficiently under different salts concentration and at different pH levels.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Ahmed et al. 2009b source

  • May combine two claims.

    Interpretationcitation

    Dubiumin's casein hydrolysis property could be useful in the dairy industry for milk clotting, as an alternative or with calf rennet, and for accelerating cheese ripening to reduce time and costs of storage and maturation.

    “This property could be useful in the dairy industry both for milk clotting, as an alternative or with calf rennet, and for the acceleration of cheese ripening to reduce the time and costs of storage and maturation (Ahmed et al. 2010).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Ahmed et al. 2010 source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Onopordosin has pI 4.4.

    “It is an aspartic protease of optimum pH 2.5 and pI 4.4.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Brutti et al. 2012 source

  • Classified as probably not a substantial claim.

    Interpretationnone given

    Different authors have used different methods and different units, making it difficult to compare the milk-clotting activity of different plant coagulants.

    “Different authors have used different methods and different units which makes it difficult to compare the milk-clotting activity of different plant coagulants.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    Sensory quality of cheeses made with onopordosin was similar to that of two commercial cheeses, but with some differential characteristics.

    “The sensory quality of the cheeses made with onopordosin compared with other two commercial cheeses was similar to that of the latter, but with some differential characteristics.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Brutti et al. 2012 source

  • Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

    Interpretationnone given

    Crude extracts can be further purified to obtain partially purified or pure enzyme depending upon the degree of purification.

    “The crude extracts can be further purified to obtain partially purified or pure enzyme depending upon the degree of purification.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationcitation

    Onopordosin is a suitable plant coagulant that can be used alone or in combination with a commercial starter to assure limited proteolysis.

    “It was concluded that onopordosin was a suitable plant coagulant which can be used alone or in combination with a commercial starter to assure a limited proteolysis (Brutti et al. 2012).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Brutti et al. 2012 source

  • May combine two claims.

    Interpretationnone given

    Plant milk-clotting enzymes differ in molecular structure and proteolytic activity.

    “These enzymes do, however, differ in molecular structure and in proteolytic activity, and there are still reports that cheese yield and cheese quality is negatively affected.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    Sunflower seed extract hydrolyzes κ-casein at the Phe105-Met106 bond, as does chymosin.

    “Mass spectrometry analysis showed that the sunflower extract hydrolyzed κ-casein at the Phe105-Met106 bond, as does chymosin.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Egito et al. 2007 source

  • Probably not a substantial claim: may not pass S2 (checkable), S3 (load-bearing), S4 (based). May combine two claims.

    Interpretationnone given

    Understanding of the action of enzymes during κ-casein cleavage and subsequent milk coagulation has increased substantially and is still ongoing.

    “The understanding of the action of the enzymes during κ-casein cleavage and subsequent milk coagulation has increased substantially, and is still going on.”
    Quote in the paperNot a substantial claim

  • Classified as probably not a substantial claim. May combine two claims.

    Interpretationnone given

    Plant proteases have been isolated from several plant sources and studied for milk-clotting ability.

    “Milk coagulants are essential for the production of cheese and plant proteases have been isolated from several plant sources and studied for milk-clotting ability.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    Albizia extract displays activity on κ-casein, with preferred target Lys116-Thr117 bond.

    “The albizia extract also displayed activity on κ-casein, but the Lys116-Thr117 bond was its preferred target (Egito et al. 2007).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Egito et al. 2007 source

  • Classified as probably not a substantial claim.

    Interpretationnone given

    Milk-clotting plant proteases can be obtained from their natural sources or through in vitro culture.

    “These enzymes can be obtained from their natural sources or through in vitro culture.”
    Quote in the paperSubstantial claim
47 of 47

  • The paper relies on this without support.

    Plant proteases have been used as milk coagulants in cheesemaking since ancient times or for centuries.

    “Plant proteases have been used as milk coagulants in cheesemaking for centuries either as crude extracts or in purified form.”
    “Plant extracts have been used as milk coagulants in cheesemaking since ancient times.”

  • The paper relies on this without support.

    The most significant difference between sheep and cow milk is protein content, an important component directly involved in coagulation.

    “The most significant difference between sheep and cow milk is protein content, an important component directly involved in coagulation.”

  • The paper relies on this without support.

    Milk-clotting activity is the most important property of enzymes used in cheesemaking.

    “Milk-clotting activity is the most important property of enzymes used in cheesemaking.”

  • The paper relies on this without support.

    A good milk-clotting enzyme is characterized by high specific caseinolytic activity and low general proteolytic activity.

    “A good milk-clotting enzyme is characterized by a high specific caseinolytic activity and a low general proteolytic activity, since the proteolysis strongly affects the sensory properties of cheese.”
    “Coagulation studies are important in the manufacture and maturation of cheeses and a good milk-clotting enzyme is characterized by a high specific caseinolytic activity and a low general proteolytic activity, since the proteolysis strongly affects the textural and sensory properties of cheese.”

  • The paper relies on this without support.

    One of the most adequate and expeditious methods to monitor proteolytic processes is polyacrylamide gel electrophoresis.

    “One of the most adequate and expeditious methods to monitor proteolytic processes is polyacrylamide gel electrophoresis.”

  • The paper relies on this without support.

    The large number of methods, different units at different conditions used by different authors has made it difficult to compare the units for milk-clotting activity.

    “The large number of methods, different units at different conditions used by different authors has made it difficult to compare the units for milk-clotting activity.”

  • The paper relies on this without support.

    Aspartic proteases have two aspartic residues at their catalytic site.

    “Aspartic proteases have two aspartic residues at their catalytic site.”

  • The paper relies on this without support.

    Cysteine proteases have great potential in food, biotechnology, and pharmaceutical industries owing to their property of being active over a wide range of temperature and pH.

    “Cysteine proteases have great potential in the food, biotechnology, and pharmaceutical industries owing to their property of being active over a wide range of temperature and pH.”

  • The paper relies on this without support.

    Serine proteases possess a serine residue in their active site and share a number of biochemical and physiological features.

    “Serine proteases possess a serine residue in their active site and share a number of biochemical and physiological features.”

  • The paper relies on this without support.

    Plant serine proteases have been found and extracted from latex, seeds, flowers, stems, leaves and roots.

    “Plant serine proteases have been found and extracted from latex, seeds, flowers, stems, leaves and roots.”

  • The paper relies on this without support.

    Proteases used as milk coagulants have been identified and studied from almost every plant part whether seed, flower, or latex.

    “Proteases used as milk coagulants have been identified and studied from almost every plant part whether it may be seed, flower, or latex.”

  • The paper relies on this without support.

    Generally, these enzymes have been extracted from their natural source by aqueous maceration of various plant organs such as flowers, seeds, roots and leaves.

    “Generally, these enzymes have been extracted from their natural source by aqueous maceration of various plant organs such as flowers, seeds, roots and leaves.”

  • The paper relies on this without support.

    In vitro production of enzymes has the potential of overcoming the low enzyme yield and difficulties in extraction from the natural sources.

    “In vitro production of enzymes has the potential of overcoming the low enzyme yield and difficulties in extraction from the natural sources.”

  • The paper relies on this without support.

    Plant extracts as a coagulant for cheesemaking has been largely used for the production of hard and semi-hard cheese products from ovine and caprine milk.

    “Plant extracts as a coagulant for cheesemaking has been largely used for the production of hard and semi-hard cheese products from ovine and caprine milk.”

  • The paper relies on this without support.

    Ultrafiltration (UF) is an established process for manipulating the protein level in cheese milk.

    “Ultrafiltration (UF) is an established process for manipulating the protein level in cheese milk.”

  • The paper relies on this without support.

    Some authors have compared the milk-clotting activity of plant proteases with calf rennet in the same conditions.

    “However, some authors have compared the milk-clotting activity of plant proteases with calf rennet in the same conditions.”

  • The paper relies on this without support.

    Milk coagulants are essential for the production of cheese.

    “Milk coagulants are essential for the production of cheese and plant proteases have been isolated from several plant sources and studied for milk-clotting ability.”

  • The paper relies on this without support.

    Extraction of milk-clotting proteases from intact plant parts is labor intensive.

    “Extraction of milk-clotting proteases from intact plants parts is labor intensive and so, plant cell cultures (in vitro) is a viable alternative to obtain clotting enzymes.”

  • The paper relies on this without support.

    Coagulation studies are important in the manufacture and maturation of cheeses.

    “Coagulation studies are important in the manufacture and maturation of cheeses and a good milk-clotting enzyme is characterized by a high specific caseinolytic activity and a low general proteolytic activity, since the proteolysis strongly affects the textural and sensory properties of cheese.”

  • The paper relies on this without support.

    Proteolysis strongly affects the textural and sensory properties of cheese.

    “Coagulation studies are important in the manufacture and maturation of cheeses and a good milk-clotting enzyme is characterized by a high specific caseinolytic activity and a low general proteolytic activity, since the proteolysis strongly affects the textural and sensory properties of cheese.”
20 of 20

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

Each contradiction between two passages listed here was confirmed by an independent check.

  • Ficus racemosa protease is classified in two different groups

    Minor

    The source list places the Ficus racemosa enzyme cited to Devaraj et al. (2008) among cysteine proteases, but the cited reference describes a protease from that source as aspartic.

1 of 1

Cites

Flagged first, then by citations

Research works printed but not linked: 1

  1. Harboe M, Broe ML, Qvist KB (2010) In: Law BA, Tamime AY (eds) Technology of cheese making. Wiley-Blackwell, London

Cited by

Sorted by citations

Related

Authors
Shah, M. A., Mir, S. A., & Paray, M. A.
Affiliations
Islamic University of Science and Technology (IN)
Corresponding author
Shah, Manzoor Ahmad
ORCID
Mir, Shabir Ahmad
Venue
Dairy Science & Technology
Publisher
Springer Science and Business Media LLC
Published
2013-07-19
Volume
94
Issue
1
Pages
5-16
ISSN
  • 1958-5586 (ISSN-L)
  • 1958-5594
DOI
10.1007/s13594-013-0144-3
OpenAlex
W2036403160
Semantic Scholar
907e08d20a318f0d4ddfa3c59907120db3c412eb
Cited by
209
References
74
Citations by year
2026: 7 · 2025: 14 · 2024: 17 · 2023: 27 · 2022: 24 · 2021: 25 · 2020: 13 · 2019: 25 · 2018: 20 · 2017: 17 · 2016: 12 · 2015: 3 · 2014: 5
FWCI
5.86
Type
journal article
Language
en
Open access
Licence
Locations
  • Dairy Science and Technology (publishedVersion, other-oa)
  • HAL (submittedVersion, other-oa)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biotechnology › Enzyme Production and Characterization
  • Life Sciences › Agricultural and Biological Sciences › Food Science › Proteins in Food Systems
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pineapple and bromelain studies
Keywords
plant proteases, milk coagulants, cheese production, aspartic protease, cysteine protease, serine protease, cheese yield

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-10-11.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-11.

Cited sources are checked by gpt-6-sol.

Statistics and arithmetic are recomputed, not judged.

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