DOCSLIB.ORG

Extraction and Immobilization of Vegetable Aspartic Proteases for Cheese Making

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Extraction and Immobilization of Vegetable Aspartic Proteases for Cheese Making CORE Metadata, citation and similar papers at core.ac.uk Provided by Università degli Studi di Napoli Federico Il Open Archive EXTRACTION AND IMMOBILIZATION OF VEGETABLE ASPARTIC PROTEASES FOR CHEESE MAKING Marilena Esposito Dottorato in Scienze Biotecnologiche – XXVII ciclo Indirizzo Biotecnologie per le Produzioni Vegetali Università di Napoli Federico II Dottorato in Scienze Biotecnologiche – XXVII ciclo Indirizzo Biotecnologie per le Produzioni Vegetali Università di Napoli Federico II EXTRACTION AND IMMOBILIZATION OF VEGETABLE ASPARTIC PROTEASES FOR CHEESE MAKING Marilena Esposito Dottorando: Marilena Esposito Relatore: Prof. Prospero Di Pierro Coordinatore: Prof. Giovanni Sannia Ora dunque rimangono queste tre cose: la fede, la speranza e la carità. Ma la più grande di tutte è la carità. 1 Corinzi 13:13 INDEX SUMMARY ............................................................................................................................................. III RIASSUNTO ........................................................................................................................................... V 1. Introduzione..................................................................................................................................... V 2. Risultati ........................................................................................................................................... VI Estrazione di attività coagulante dalle foglie di carciofo (Cynara scolymus) ................................. VI Estrazione di attività coagulante dai fiori di cardo (Carduus defloratus) ...................................... VII Immobilizzazione di enzimi con attività coagulante ..................................................................... VIII 1. INTRODUCTION .................................................................................................... 1 1.1 Enzyme global market situation .................................................................................................... 3 1.2 Proteases ...................................................................................................................................... 3 1.2.1 Aspartic proteases ................................................................................................................. 4 1.2.2 Milk clotting aspartic proteases.............................................................................................. 5 1.2.3 Plant proteases as milk clotting enzyme for cheese making ................................................. 6 1.3 Overview of enzyme immobilization .............................................................................................. 7 1.3.1 Epoxy support ........................................................................................................................ 9 1.4 Aim of the thesis .......................................................................................................................... 10 1.5 References .................................................................................................................................. 10 2. RESULTS ............................................................................................................. 15 2.1 Extraction and characterization of milk clotting proteases from artichoke (Cynara scolymus, L.) leaves and alpine thistle (Carduus defloratus subsp. carlinifolius, Lam.) flowers ................. 17 2.1.1 Introduction .............................................................................................................................. 17 2.1.2 Materials and methods ............................................................................................................. 18 2.1.2.1 Materials ........................................................................................................................... 18 2.1.2.2 Methods ............................................................................................................................ 18 2.1.2.2.1 Protein extraction ........................................................................................................... 18 2.1.2.2.2 SDS-PAGE analysis ...................................................................................................... 19 2.1.2.2.3 Two-dimensional electrophoresis (2DE) ........................................................................ 19 2.1.2.2.4 Purification steps ........................................................................................................... 19 2.1.2.2.5 Milk-clotting activity ........................................................................................................ 19 2.1.2.2.6 Inhibition studies ............................................................................................................ 20 2.1.2.2.7 Statistical analysis ......................................................................................................... 20 2.1.3 Results and Discussion ............................................................................................................ 20 2.1.3.1 Cynara scolymus .............................................................................................................. 20 2.1.3.2 Carduus defloratus subsp. carlinifolius, Lam. ................................................................... 25 2.1.4 Conclusions .............................................................................................................................. 27 2.1.5 References ............................................................................................................................... 28 2.2 Immobilization of aspartic proteases with milk-clotting activity from Mucor miehei and Carduus defloratus L. subsp. carlinifolius ....................................................................................... 30 2.2.1 Introduction .............................................................................................................................. 30 2.2.2 Materials and Methods ............................................................................................................. 31 2.2.2.1 Materials ........................................................................................................................... 31 2.2.2.2 Methods ............................................................................................................................ 31 2.2.2.2.1 Enzyme purification ....................................................................................................... 31 2.2.2.2.2 Immobilization procedure ............................................................................................... 31 2.2.2.2.3 Determination of protein concentration .......................................................................... 32 2.2.2.2.4 Clotting activity assay .................................................................................................... 32 2.2.2.2.5 Determination of immobilized activity ............................................................................ 32 2.2.2.2.6 Reusability of the immobilized enzymes ........................................................................ 32 2.2.2.2.7 Statistical analysis ......................................................................................................... 33 I 2.2.3 Results and Discussion ............................................................................................................ 33 2.2.3.1 Mucor miehei protease immobilization ............................................................................. 33 2.2.3.2 Carduus defloratus protease immobilization .................................................................... 36 2.2.4 Conclusions .............................................................................................................................. 38 2.2.5 References ............................................................................................................................... 39 3. CONCLUSIONS ................................................................................................... 41 Appendix .............................................................................................................................................. 45 ACKNOWLEDGMENTS ....................................................................................................................... 46 II SUMMARY The worldwide increase in cheese production and consumption, together with the reduced supply and consequent price increase of calf rennet, has led to the search of new milk coagulant agents. Much research interest has been thus directed towards discovering milk clotting enzymes which would satisfactory replace calf rennet in cheese manufacture. Microbial rennet produced by genetically engineered bacteria has proven to be a suitable substitute even though the consumer constraints on the use of animal rennet either for religion reasons (Judaism and Islam) or diet habits (vegetarianism) or consumer concerns regarding genetically engineered foods (Germany, Netherlands and France forbid the use of recombinant calf rennet) have led to a growing interest toward vegetable coagulants. Therefore, several plant sources of milk-clotting enzymes have been investigated. Unfortunately, most of the “plant rennets” were found unsuitable because they produced extremely bitter dairy products due to a strong proteolysis.
Load more

Recommended publications
  • Enzymes Handling/Processing
    Enzymes Handling/Processing 1 Identification of Petitioned Substance 2 3 This Technical Report addresses enzymes used in used in food processing (handling), which are 4 traditionally derived from various biological sources that include microorganisms (i.e., fungi and 5 bacteria), plants, and animals. Approximately 19 enzyme types are used in organic food processing, from 6 at least 72 different sources (e.g., strains of bacteria) (ETA, 2004). In this Technical Report, information is 7 provided about animal, microbial, and plant-derived enzymes generally, and more detailed information 8 is presented for at least one model enzyme in each group. 9 10 Enzymes Derived from Animal Sources: 11 Commonly used animal-derived enzymes include animal lipase, bovine liver catalase, egg white 12 lysozyme, pancreatin, pepsin, rennet, and trypsin. The model enzyme is rennet. Additional details are 13 also provided for egg white lysozyme. 14 15 Chemical Name: Trade Name: 16 Rennet (animal-derived) Rennet 17 18 Other Names: CAS Number: 19 Bovine rennet 9001-98-3 20 Rennin 25 21 Chymosin 26 Other Codes: 22 Prorennin 27 Enzyme Commission number: 3.4.23.4 23 Rennase 28 24 29 30 31 Chemical Name: CAS Number: 32 Peptidoglycan N-acetylmuramoylhydrolase 9001-63-2 33 34 Other Name: Other Codes: 35 Muramidase Enzyme Commission number: 3.2.1.17 36 37 Trade Name: 38 Egg white lysozyme 39 40 Enzymes Derived from Plant Sources: 41 Commonly used plant-derived enzymes include bromelain, papain, chinitase, plant-derived phytases, and 42 ficin. The model enzyme is bromelain.
    [Show full text]
  • The Sysco Cheese Product Catalog
    > the Sysco Cheese Product Catalog Sysco_Cheese_Cat.indd 1 7/27/12 10:55 AM 5 what’s inside! 4 More Cheese, Please! Sysco Cheese Brands 6 Cheese Trends and Facts Creamy and delicious, 8 Building Blocks... cheese fi ts in with meal of Natural Cheese segments during any Blocks and Shreds time of day – breakfast, Smoked Bacon & Cheddar Twice- Baked Potatoes brunch, lunch, hors d’oeuvres, dinner and 10 Natural Cheese from dessert. From a simple Mild to Sharp Cheddar, Monterey Jack garnish to the basis of and Swiss a rich sauce, cheese is an essential ingredient 9 10 12 A Guide to Great Italian Cheeses Soft, Semi-Soft and for many food service Hard Italian Cheeses operations. 14 Mozzarella... The Quintessential Italian Cheese Slices, shreds, loaves Harvest Vegetable French and wheels… with Bread Pizza such a multitude of 16 Cream Cheese Dreams culinary applications, 15 16 Flavors, Forms and Sizes the wide selection Blueberry Stuff ed French Toast of cheeses at Sysco 20 The Number One Cheese will provide endless on Burgers opportunities for Process Cheese Slices and Loaves menu innovation Stuff ed Burgers and increased 24 Hispanic-Style Cheeses perceived value. Queso Seguro, Special Melt and 20 Nacho Blend Easy Cheese Dip 25 What is Speciality Cheese? Brie, Muenster, Havarti and Fontina Baked Brie with Pecans 28 Firm/Hard Speciality Cheese Gruyère and Gouda 28 Gourmet White Mac & Cheese 30 Fresh and Blue Cheeses Feta, Goat Cheese, Blue Cheese and Gorgonzola Portofi no Salad with 2 Thyme Vinaigrette Sysco_Cheese_Cat.indd 2 7/27/12 10:56 AM welcome.
    [Show full text]
  • Progress in the Field of Aspartic Proteinases in Cheese Manufacturing
    Progress in the field of aspartic proteinases in cheese manufacturing: structures, functions, catalytic mechanism, inhibition, and engineering Sirma Yegin, Peter Dekker To cite this version: Sirma Yegin, Peter Dekker. Progress in the field of aspartic proteinases in cheese manufacturing: structures, functions, catalytic mechanism, inhibition, and engineering. Dairy Science & Technology, EDP sciences/Springer, 2013, 93 (6), pp.565-594. 10.1007/s13594-013-0137-2. hal-01201447 HAL Id: hal-01201447 https://hal.archives-ouvertes.fr/hal-01201447 Submitted on 17 Sep 2015 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Dairy Sci. & Technol. (2013) 93:565–594 DOI 10.1007/s13594-013-0137-2 REVIEW PAPER Progress in the field of aspartic proteinases in cheese manufacturing: structures, functions, catalytic mechanism, inhibition, and engineering Sirma Yegin & Peter Dekker Received: 25 February 2013 /Revised: 16 May 2013 /Accepted: 21 May 2013 / Published online: 27 June 2013 # INRA and Springer-Verlag France 2013 Abstract Aspartic proteinases are an important class of proteinases which are widely used as milk-coagulating agents in industrial cheese production. They are available from a wide range of sources including mammals, plants, and microorganisms.
    [Show full text]
  • Of 3 Enzymatic Assay of RENNIN (EC 3.4.23.4) PRINCIPLE
    Enzymatic Assay of RENNIN (EC 3.4.23.4) PRINCIPLE: Rennin 1 Milk > Clotted Milk CONDITIONS: T = 30°C METHOD: Clotting time REAGENTS: A. 1 M Calcium Chloride Solution (Prepare 5 ml in deionized water using Calcium Chloride, Dihydrate, Sigma Prod. No. C-3881.) B. 10.4% (w/v) Milk Solution with 10 mM Calcium Chloride (Milk) (Prepare by dissolving 20.9 g of Carnation Instant Nonfat Dry Milk Powder in 200 ml of deionized water or use skimmed cow's milk. Then add 2 ml of Reagent A.) C. 0.1% (w/v) Rennet Standard Solution (Rennet Std) (Prepare 10 ml in deionized water using Rennet, Sigma Prod. No. R-3376.) D. Rennin Enzyme Solution (Rennin) (Immediately before use, prepare a solution containing 0.01 - 0.05 mg/ml of Rennin in cold deionized water. Dilute accordingly so that the clotting time is 0.75 - 1.5 times that of Reagent C.) VEMILK01 Page 1 of 3 Revised: 08/29/97 Enzymatic Assay of RENNIN (EC 3.4.23.4) PROCEDURE: Step 1: Pipette (in milliliters) the following reagents into a 50 ml Erlenmeyer flask: Control Reagent B (Milk) 10.00 Incubate at 30°C in a water bath for 45 minutes. At t0 add: Reagent C (Rennet Std) 1.00 Swirl gently (Erlenmeyer flask) at 30°C in a water bath. Stop timing and swirling when a white-translucent semi- liquified film appears on the side of the flask above the milk. This is t1. After t1, the milk will continue to congeal. Step 2: Pipette (in milliliters) the following reagents into a suitable test tube.
    [Show full text]
  • An Evaluation of the Clotting Properties of Three Plant Rennets in the Milks of Different Animal Species
    foods Article An Evaluation of the Clotting Properties of Three Plant Rennets in the Milks of Different Animal Species Katia Liburdi 1,* , Carlo Boselli 2, Gilberto Giangolini 2, Simonetta Amatiste 2 and Marco Esti 1 1 Department of Agricultural and Forestry Sciences (DAFNE), Tuscia University, Via San Camillo de Lellis, 01100 Viterbo, Italy; [email protected] 2 Experimental Zooprophylactic Institute Lazio and Toscana “Mariano Aleandri”, Via Appia Nuova 1411, 00178 Rome, Italy; [email protected] (C.B.); [email protected] (G.G.); [email protected] (S.A.) * Correspondence: [email protected] Received: 8 October 2019; Accepted: 16 November 2019; Published: 20 November 2019 Abstract: Cynara cardunculus, Carica papaya and Ficus carica extracts are proposed as milk coagulants herein. Their coagulation efficiency was measured in bovine, buffalo, goat and sheep milk incubated at different temperatures. The milk-clotting and proteolytic activities as well as the lactodynamographic parameters were determined considering animal rennet as a reference coagulant. The vegetable coagulant, extracted from C. cardunculus pistils, proved to be the most suitable milk-clotting enzyme for cheesemaking, since it possesses similar milk clotting properties to conventional calf rennet. F. carica latex, but seemed to be a promising alternative coagulant at higher temperatures. The strong proteolytic activity of papain caused poor milk coagulation in all milk samples. To conclude, this result also supports the original hypothesis of this study that the excessive proteolytic nature of plant coagulants can negatively affect the cheesemaking process. The optimization of using a plant rennet in a dairy application can be done by selecting the appropriate plant rennet with a consistent clotting efficiency.
    [Show full text]
  • Agricultural Marketing Service, USDA § 58.443
    Agricultural Marketing Service, USDA § 58.443 components in cheese shall have a cured for a period of 60 days at a tem- pleasing and desirable taste and odor perature not less than 35 °F. If the milk and shall have the ability to actively is held more than 2 hours between time produce the desired results in the of receipt or heat treatment and set- cheese during the manufacturing proc- ting, it shall be cooled to 45 °F. or ess. lower until time of setting. § 58.434 Calcium chloride. § 58.440 Make schedule. Calcium chloride, when used, shall A uniform schedule should be estab- meet the requirements of the Food lished and followed as closely as pos- Chemical Codex. sible for the various steps of setting, cutting, cooking, draining the whey § 58.435 Color. and milling the curd, to promote a uni- Coloring when used, shall be Annatto form quality of cheese. or any cheese or butter color which meet the requirements of the Food and § 58.441 Records. Drug Administration. Starter and make records should be kept at least three months. § 58.436 Rennet, pepsin, other milk clotting enzymes and flavor en- (Approved by the Office of Management and zymes. Budget under OMB control number 0583– 0047) 1 Enzyme preparations used in the manufacture of cheese shall be safe and [40 FR 47911, Oct. 10, 1975. Redesignated at 42 suitable. FR 32514, June 27, 1977, and further redesig- nated at 46 FR 63203, Dec. 31, 1981, as amend- § 58.437 Salt. ed at 47 FR 745, Jan. 7, 1982] The salt shall be free-flowing, white § 58.442 Laboratory and quality con- refined sodium chloride and shall meet trol tests.
    [Show full text]
  • Efficacy of Chicken Pepsin As a Milk Clotting Enzyme 1
    684 Journal o{Food Protection Vol. 41, No.9. Pages 684-688 (September. 1978) Copyright 1978, International Association of Milk, Food, and Environmental Sanitarians Efficacy of Chicken Pepsin as a Milk Clotting Enzyme 1 S. GORDIN and I. ROSENTHAL* Dairy Laboratory, Division ofFood Technology Agricultural Research Organization, The Volcani Center Downloaded from http://meridian.allenpress.com/jfp/article-pdf/41/9/684/1650145/0362-028x-41_9_684.pdf by guest on 26 September 2021 P.O. Box 6, Bet Dagan, (20-500), Israel (Received for publication December 19, 1977) ABSTRACT Enzyme activity assays Comparative laboratory tests of cheesemaking show similarity Enzyme activity was estimated by the milk clotting test done in test tubes periodically rotated in a thermostatic bath at 30 C, unless between chicken pepsin and calf rennet. Suitability of chicken pepsin othernise specified. To ensure reproducible results, a reconstituted for large-scale production of Emmental (Swiss) and Kashkaval-type skimmed milk known as "Berridge substrate" was employed. This cheeses was tested. substrate consists of 12 g of low-heat spray-dried skim milk powder (6 mg of whey protein/g index) dissolved in 100 ml of 0,01 M CaC1 2 solution. After adding the skim milk powder to the CaC12 solution, the mixture was stirred for 20 min and left to stand at room temperature Use of calf rennet as a milk clotting enzyme in the for an additional hour. This preparation procedure was adopted since manufacture of cheese has been predominant in the we noted that the time required for coagulation increased with the age of "Berridge substrate", particularly for freshly prepared milk industry for centuries.
    [Show full text]
  • Types of Cheeses
    Types of Cheeses Asiago cheese is a nutty flavored cheese that hails from Europe. It is named for a region in Italy where it was first produced. This region is known as the Asiago High Plateau, which lies within the Italian Alps. Asiago cheese is produced in two forms as follows: fresh Asiago, also known as Pressato, and mature Asiago, which is called Asiago d´Allevo. Fresh Asiago has an off-white color and is milder in flavor than mature asiago. Mature asiago also has a more yellowish color and is somewhat grainy in texture. Blue cheese is a general classification of cow's milk, sheep's milk, or goat's milk cheeses that have had Penicillium cultures added so that the final product is spotted or veined throughout with blue, blue-gray or blue-green mold, and carries a distinct smell. Some blue cheeses are injected with spores before the curds form and others have spores mixed in with the curds after they form. Blue cheese was initially produced in caves Blue cheeses are typically aged in a temperature- controlled environment such as a cave. The characteristic flavor of blue cheeses tends to be sharp and a bit salty. Due to this strong flavor and smell, blue cheeses are often considered an acquired taste. They can be eaten by themselves or can be crumbled or melted over foods. Cheddar cheese originated in the village of Cheddar, England. A firm, cow's milk cheese that ranges in flavor from mild to sharp and in color from a natural white to pumpkin orange.
    [Show full text]
  • Molecular Cloning and Characterization of Procirsin, an Active Aspartic Protease Precursor from Cirsium Vulgare (Asteraceae)
    Phytochemistry 81 (2012) 7–18 Contents lists available at SciVerse ScienceDirect Phytochemistry journal homepage: www.elsevier.com/locate/phytochem Molecular cloning and characterization of procirsin, an active aspartic protease precursor from Cirsium vulgare (Asteraceae) Daniela Lufrano a, Rosário Faro b, Pedro Castanheira c, Gustavo Parisi d, Paula Veríssimo b,e, ⇑ Sandra Vairo-Cavalli a, Isaura Simões b,c, , Carlos Faro b,c,e a Laboratorio de Investigación de Proteínas Vegetales (LIPROVE), Departamento de Ciencias Biológicas, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, C.C. 711, 1900 La Plata, Argentina b CNC-Center for Neuroscience and Cell Biology, University of Coimbra, 3004-517 Coimbra, Portugal c Biocant, Biotechnology Innovation Center, 3060-197 Cantanhede, Portugal d Departamento de Ciencia y Tecnología, Universidad Nacional de Quilmes, Roque Sáenz Peña 352, Bernal, Buenos Aires, B1876BXD, Argentina e Department of Life Sciences, University of Coimbra, 3004-517 Coimbra, Portugal article info abstract Article history: Typical aspartic proteinases from plants of the Astereaceae family like cardosins and cyprosins are well- Received 16 February 2012 known milk-clotting enzymes. Their effectiveness in cheesemaking has encouraged several studies on Received in revised form 23 May 2012 other Astereaceae plant species for identification of new vegetable rennets. Here we report on the clon- Available online 21 June 2012 ing, expression and characterization of a novel aspartic proteinase precursor from the flowers of Cirsium vulgare (Savi) Ten. The isolated cDNA encoded a protein product with 509 amino acids, termed cirsin, Keywords: with the characteristic primary structure organization of plant typical aspartic proteinases. The pro form Cirsium vulgare of cirsin was expressed in Escherichia coli and shown to be active without autocatalytically cleaving its Asteraceae pro domain.
    [Show full text]
  • US7553632.Pdf
    US007553632B2 (12) United States Patent (10) Patent No.: US 7,553,632 B2 Niles et al. (45) Date of Patent: Jun. 30, 2009 (54) LUMINOGENIC AND NONLUMINOGENIC WO WO-020O882 A2 1, 2002 MULTIPLEX ASSAY WO WO-0212547 A1 2, 2002 WO WO-O3O25192 A2 3, 2003 (75) Inventors: Andrew Niles, Madison, WI (US); Terry W w858. A. 3. L. Riss, Oregon, WI (US) WO WO-2005073722 A3 8, 2005 (73) Assignee: Promega Corporation, Madison, WI WO WO-2007027653 A1 3f2007 (US) OTHER PUBLICATIONS (*) Notice: Subject to any disclaimer, the term of this O'Connellet al. Live Dead Assay for Cell Viability:AfCS Procedure patent is extended or adjusted under 35 EEEata/cgi-bin/Protocol File.cgi/afcs hisygaling-gateway.org U.S.C. 154(b) by 69 days. pdf?pid=PP00000023 downloaded Nov. 19, 2007.* Rosser et al. Calpain Activity Increases in Hepatocytes Following (21) Appl. No.: 11/489,978 Addition of ATP; The Journal of Biological Chemistry, vol. 268, No. 1-1. 31 (1993) pp. 23.593-23600.* (22) Filed: Jul. 20, 2006 “Partial International Search Report for corresponding PCT Appli cation No. PCT/US2005/002158”. (Oct. 12, 2006), 2 pgs. (65) Prior Publication Data Grant, S. K., et al., “Development of Novel Assays for Proteolytic Enzymes Using Rhodamine-Based Fluorogenic Substrates”,Journal US 2007/0178545 A1 Aug. 2, 2007 of Biomolecular Screening, 7(6), (2002), 531-540. Related U.S. ApplicationO O Data "ApoptosisFCE Inducers 2(8), Abstract And The No. Assay F. of FR Caspase ActivityANA Biomol', Stud (63) Continuation of application No. PCT/US2005/ ies for Experimental Biology (Apr.
    [Show full text]
  • The Iushrut of Microbial Enzymes
    YD 87:10.1994 THE IUSHRUT OF MICROBIAL ENZYMES Rabbi Kassel Abelson 77zis paper was approved by the CJT.S on December 71, 7991, by a vote of' sixteen in Javor and three abstainifl{'· (16-0-:J). WJting infiwor: Rabbis Kassel Alwlson, Ben Lion Bergman, Stanley Bmmnick, L'lliot N. Dorff, Samuel Fmint, ]an Cntyl Kaufman, Judah Kugen, H'rrwn R. Kurtz, Alan n. runts, Paul Plothin, r-1-vram Tsrael Reisnet~ Joel K Rernbaum, Joel Hoth, Gerald Slrolniii:, Cordon '/lu:lrer, wul Gerald Yelizer. Abstai.nin~;: Rabbi.s Jerome /H. Ljxstein, Susan Cros.sman, and Aaron T.. Mackler. 17w Committee on ]m;ish T.aw and StandanLs rijthe Rabbinical Assembly provides guidance in matters rij halahhahfor the Conservative movement. The individual rabhil however, is the authorif:yj(Jr the interpretation and application of all matters of'lwlaklwh. Are cheeses made with microbial milk-clotting enzymes (sometimes referred to as rennin or rennet) kosher? Rabbi Isaac Klein, of blessed memory, in a responsum adopted unanimously by the CJLS on January 20, 1972, affirmed, "It is our considered opinion that cheeses, all cheeses, including those in which rennet, any rennet, is used as the curdling agent should be per­ mitted." And he explains: TI1e rennet used today cannot be considered forbidden because, first of all, most of it is derived from dried up skins that have become like a piece of wood. In addition, the extraction is brought about by the use of strong chemicals and acids which removes the substance from the status of a food fit even for a dog.
    [Show full text]
  • Calf Rennet Production and Its Performance Optimization
    Journal of Applied and Natural Science 10(1): 247 - 252 (2018) Calf rennet production and its performance optimization Asfawosen Mamo and Nedumaran Balasubramanian* Chemical Engineering Program, School of Mechanical, Chemical and Materials Engineering, Adama Science and Technology University, POBox 1888, Adama, Ethiopia *Corresponding author. E-mail: [email protected] Received: August 26, 2017; Revised received: October 10, 2017; Accepted: January 31, 2018 Abstract: Milk clotting enzymes are one of the most significant cheese making raw materials impacting and regulat- ing milk coagulation activity. Calf rennet is one of the enzymes which extracted from suckling calf abomasum. The rennet extracted was optimized for its clotting time t (s) by varying three levels of temperature (30, 35 and 40 oC), pH (4, 5 and 6) and CaCl2 concentration (0.1, 0.2 and 0.3 gm/500 ml of milk) using response surface method (RSM) and also its milk clotting activity (MCA) was determined. Based on the optimized result, temperature 39.13oC, pH 4 and CaCl2 concentration of 0.21 gm/500ml had resulted minimum clotting time of 91.27 s. Using this minimum clot- ting time, the strength was found to be 1: 13148, establishing the fact that the calf rennet is the best natural coagu- lant of milk. Keywords: Abomasum, Calf rennet, Clotting time, Response surface method. INTRODUCTION pre-weaned calves (Moran, 2005). Milk coagulation properties (MCP) are an important The products obtained from Cows are well known to aspect in assessing cheese-making ability. Several mankind, such as Gorochan (Sailaja et al., 2017), studies showed that favorable conditions of milk reac- known as Gorojanam (Sanskrit, Tamil) is one of the tivity with rennet, curd formation rate, and curd products of cholelithiasis in the gall bladder and ducts strength, as well as curd synthesis, have a positive ef- of cow and bull which is laxative, antispasmodic, cho- fect on the entire cheese-making process and subse- lagogue and cooling and used as traditional Indian quently on the ripening of cheese.
    [Show full text]