Valorisation of microbial biodiversity: Characterization of a milk-clotting enzyme produced by Bacillus velezensis FK6A strain isolated from Algerian dairy farm soil in light of its use in the manufacture of hard cooked cheeses.

F. Boumediene, S. Talantikite, S. Haddad, A. Nouani, S. Triki, S. Belbraouet

Abstract


Abstract: A strain known as FK6A, which produces a milk-clotting enzyme isolated from the soil of an Algerian dairy factory, was identified as Bacillus velezensis based on morphology and internal transcription spacer sequence. Phylogenetic and sequencing analysis of 16S rDNA gene showed a 100% identity match between the tested strainand the Bacillusvelezensis in the database (Code MH368142). This strain produced an active extracellular protease purified by partial purification using ammonium sulfate fractionation(40–80% saturation) followed by chromatography of molecular exclusion, which revealed the existence of tow active peaks. The optimal activity was observed at pH 5.7; 75°C and 40 mM of CaCl2. The clotting enzyme has been shown to be a metalloprotease inhibited by EDTA. The highly proteolytic activity of the Bacillus velezensis extracts observed on the curds appears a few minutes after the appearance of a firm coagulum reflected by the appearance of an abundant serum.Thus, to solve this problem and to inhibit proteolytic activity, the curds were cooked after brewing during the manufacturing process.


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Jacob M.;Jaros D.;Rohm H.; Recent advances in milk clotting enzymes. International Journal of Dairy Technology 64(1) (2011) 14-33.

Mazorra-Manzano M.A.;Perea-Gutierrez T.C.;Lugo-Sanchez M.E..;Ramirez-Suarez J.C.;Torres-Llanez M.J..;Gonzalez-Cordova A.F.; Comparison of the milk-clotting properties of three plant extracts. Food Chemistry 141(3) (2013) 1902-7.

Dutt K.;Meghwanshi G.K.;Gupta P.;Saxena R.K.; Role of casein on induction and enhancement of production of a bacterial milk clotting protease from an indigenously isolated Bacillus subtilis. Letter of Applied Microbiology 46(5) (2008) 513-8.

Singh T.K.;Drake M.A.;Cadwallader K.R.; Flavor of Cheddar Cheese: A Chemical and Sensory Perspective. Comprehensive reviews in food science and food safety 2(4) (2003) 166-89.

Wesley, M.Cg.; Cheddar Cheese Flavor. A Review of Current Progress. Journal of Agriculture and Food Chemistry, 23(6) (1975) 1047-50.

Ageitos J.M.;Vallejo J.A.;Sestelo A.B.F.;Poza M.;Villa T.G.; Purification and characterization of a milk-clotting protease from Bacillus licheniformis strain USC13. J Appl Microbiol 103(6) (2007) 2205-13.

Ding Z L.S., Gu Z, Zhang L, Zhang K, Shi G. Production of milk-clotting enzyme by Bacillus subtilis B1 from wheat bran. African Journal of Biotechnology 10(46) (2011) 9370-8.

Talantikite-Kellil, S.; Nouani, A.; , Saunier, M.; Gillmann, L.; Brahmi, A.; Fazouane, F.; Isolation and identification of bacterial strain I33M producing milk-clotting enzyme: Optimization of culture parameters using response surface. African Journal of Biotechnology 11(100) (2012) 16594-606.

Rao, L.K.; Mathur ,D.K.; Studies on the production of bacterial rennet in a pilot plant fermentor. Biotechnology and bioengineering 17(9)1975) 1349-61.

Shieh, C.J.; Phan Thi, L.A.; Shih , I.L.;Milk-clotting enzymes produced by cult (ure of Bacillus subtilis natto. Biochemical Engineering Journal 43(1) (2009) 85-91.

Silva ,B.L.;Geraldes , F.M.;Murari C.S.;Gomes E..;Da-Silva R. Production and characterization of a milk-clotting protease produced in submerged fermentation by the thermophilic fungus Thermomucor indicae-seudaticae N31. Applied Biochemical and Biotechnoogyl 172(4) (2014) 1999-2011.

Lowry , O.H.; Rosebrough, N.J.;Farr ,A.L.;Randall, R.J.; Protein measurement with the Folin phenol reagent. J Biol Chem 193(1) (1951) 265-75.

Raposo ,S.; Domingos, A. ; Purification and characterization milk-clotting aspartic proteinases from Centaurea calcitrapa cell suspension cultures. Process Biochemistry 43(2) (2008) 139-44.

He, X.;Ren, F.;Guo, H.; Zhang, W.; Song, X.;Gan B.; Purification and properties of a milk-clotting enzyme produced by Bacillus amyloliquefaciens D4. Korean Journal of Chemical Engineering 28(1) (2010) 203-8.

Kahlouche , A.; Nouani ,A.; Characterization of a milk-clotting enzyme produced by Bacillus mojavensis P47M strain isolated from Algerian dairy farm soil. Research Journal of Biotechnology 12(12) (2017) 37-45.

Lemes, A.C.;Pavón ,Y.;Lazzaroni ,S.;Rozycki, S.;Brandelli, A.;Kalil, S.J.; A new milk-clotting enzyme produced by Bacillus sp. P45 applied in cream cheese development. LWT - Food Science and Technology 66 (2016) 217-24.

Khan, M.R.;Blain, J.A.;Patterson, J.D.; Partial Purification of Mucor pusillus Intracellular Proteases. Applied Environnmental and Microbiology 45(1) (1983) 94-6.

Esawy ,M.A.;Combet-Blanc ,Y.; Immobilization of Bacillus licheniformis 5A1 milk-clotting enzyme and characterization of its enzyme properties. World Journal of Microbiology and Biotechnology 22(3) (2006) 197-200.

Won Park, Y.;Kobayashi, H.;Kusakabe ,I.;Murakami, K.; Purification and Characterization of Soymilk-clotting Enzymes fromBacillussp. K-295G-7. Agricultural and Biological Chemistry 51(9) (2016) 2343-9.

Ding, Z. W.W.; Wang, B.; Ouyang , A.; Xiao, S.; Wang, Y.; Liu , S.; Ding , M.; Zhang, L.; Shi, G.;Production and characterization of milk-clotting enzyme from Bacillus amyloliquefaciens JNU002 by submerged fermentation. European Food Research and Technology 234(3) (2012) 415–21.

El-Bendary, M.A.;Moharam, M.E.;Ali, T.H.; Purification and Characterization of Milk Clotting Enzyme Produced by Bacillus sphaericus. Journal of Applied Sciences Research 3(8) (2007) 695-9.

Kumari Narwal, R.;Bhushan,B.;Pal, A.;Panwar, A.;Malhotra, S.; Purification, physico-chemico-kinetic characterization and thermal inactivation thermodynamics of milk clotting enzyme from Bacillus subtilis MTCC 10422. LWT - Food Science and Technology 65 (2016) 652-60.

Yasuda, M.;Aoyama, M.;Sakaguchi,M.;Nakachi, K.;Kobamoto, N.; Purification and characterization of a soybean-milk-coagulating enzyme from Bacillus pumilus TYO-67. Applied Microbiology and Biotechnology 51(4) (1999) 474-9.

Peng ,Y.;Yang, X.;Zhang, Y.; Microbial fibrinolytic enzymes: an overview of source, production, properties, and thrombolytic activity in vivo. Applied Microbiology and Biotechnology 69(2) (2005)126-32.

Li, Y.;Liang, S.;Zhi, D.;Chen, P.;Su, F.;Li, H.; Purification and characterization of Bacillus subtilis milk-clotting enzyme from Tibet Plateau and its potential use in yak dairy industry. European Food Research and Technology 234(4) (2012) 733-41.

Hang F..;Wang Q..;Hong Q..;Liu P..;Wu Z..;Liu, Z.; Zhang, H.; Chen, W.; Purification and characterization of a novel milk-clotting metalloproteinase from Paenibacillus spp. BD3526. International Journal of Biological Macromolecules 85 (2016) 547-54.

Ahmed S.A..;Helmy W.A. Comparative evaluation of Bacillus licheniformis 5A5 and Aloe variegata milk-clotting enzymes. Brazilian Journal of Chemical Engineering 29(1)201269 - 76. v

Mahaut, M.;Jeantet, R.;Brule, G;. Initiation à La Technologie Fromagère. LAVOISIER / TEC ET DOC. 2007.

Glantz, M.;Lindmark, Mansson, H.;Stalhammar, H.;Paulsson ,M.; Effect of polymorphisms in the leptin, leptin receptor, and acyl-coenzyme A:diacylglycerol acyltransferase 1 (DGAT1) genes and genetic polymorphism of milk proteins on cheese characteristics. Journal of Dairy Science 4(7) (2011) 3295-304.

Garnot, P.;Martin, P.; La présure: composition, activité, son rôle en fromagerie. Technique laitiere 930(3) (1980) 27-30.

Sbodio, O.A.;Tercero, E.J.;Coutaz, V.R.;Luna, J.A.;Martínez, E.; Simultaneous interaction of pH, CaCl2 addition, temperature and enzyme concentration on milk coagulation properties / Interacción simultánea del pH, CaCl 2 añadido, temperatura y concentración enzimática sobre la coagulación de la leche. Food science and technology international 3(4) (2016) 291-8.


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