An Evaluation of the Clotting Properties of Three Plant Rennets in the Milks of Different Animal Species

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. These innovative manufacturing processes may also lead to the optimization and production of new cheese varieties.

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A novel milk-clotting enzyme from Aspergillus oryzae and A. luchuensis is an aspartic endopeptidase PepE presumed to be a vacuolar enzyme

Bioscience Biotechnology and Biochemistry ◽

10.1093/bbb/zbac005 ◽

2022 ◽

Author(s):

Yoko Takyu ◽

Taro Asamura ◽

Ayako Okamoto ◽

Hiroshi Maeda ◽

Michio Takeuchi ◽

...

Keyword(s):

Aspergillus Oryzae ◽

Milk Clotting ◽

Milk Clotting Enzyme ◽

Proteolytic Activities ◽

Homologous Enzyme ◽

Traditional Fermentation ◽

Enzyme Source ◽

Cheese Production ◽

Milk Clotting Activity ◽

Aspartic Endopeptidase

Abstract Aspergillus oryzae RIB40 has 11 aspartic endopeptidase genes. We searched for milk-clotting enzymes based on the homology of the deduced amino acid sequence with chymosins. As a result, we identified a milk-clotting enzyme in A. oryzae. We expected other Aspergillus species to have a homologous enzyme with milk-clotting activity, and we found the most homologous aspartic endopeptidase from A. luchuensis had milk-clotting activity. Surprisingly, two enzymes were considered as vacuole enzymes according to a study on A. niger proteases. The two enzymes from A. oryzae and A. luchuensis cleaved a peptide between the 105Phe-106Met bond in κ-casein, similar to chymosin. Although both enzymes showed proteolytic activity using casein as a substrate, the optimum pH values for milk-clotting and proteolytic activities were different. Furthermore, the substrate specificities were highly restricted. Therefore, we expected that the Japanese traditional fermentation agent, koji, could be used as an enzyme source for cheese production.

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Studies on Milk-Clotting Enzyme from the "Litsusu" Tree (Wrightiana calysina): Evidence for Milk Coagulation

Nihon Chikusan Gakkaiho ◽

10.2508/chikusan.54.11_720 ◽

1983 ◽

Vol 54 (11) ◽

pp. 720-728

Author(s):

Akiyoshi HOSONO ◽

Hajime OTANI ◽

Fumisaburo TOKITA

Keyword(s):

Milk Clotting ◽

Milk Clotting Enzyme ◽

Milk Coagulation

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Cheese-making with a vegetable rennet from Cardo (Cynara cardunculus)

Journal of Dairy Research ◽

10.1017/s0022029900014163 ◽

1972 ◽

Vol 39 (3) ◽

pp. 335-343 ◽

Cited By ~ 59

Author(s):

F. Vieira de Sá ◽

Manuela Barbosa

Keyword(s):

Rheological Behaviour ◽

Cow’S Milk ◽

Cynara Cardunculus ◽

Cheese Making ◽

Cow's Milk ◽

Milk Clotting ◽

Ph Values ◽

Sheep Milk ◽

Milk Clotting Enzyme ◽

Cheese Curd

SummaryThe milk-clotting enzyme found in the flowers of Cardo (Cynara cardunculus) was investigated as to its suitability as a substitute for traditional animal rennet used in cheese-making. The influence of milk pH, temperature and quantity on the clotting activity of this enzyme was studied. Rheological behaviour of cow's-milk and sheep-milk curds, from renneting to cutting, was determined with the Plint cheese curd torsiometer for various Ca contents and pH values. Edam, Serra and Roquefort cheeses were made and the protein breakdown which occurred in the cheese during the ripening period was determined. Animal rennet was used as a control in all the experiments. The enzyme from Cardo was a satisfactory substitute for animal rennet with cow's milk and was even more suitable for sheep's milk. It was found to be a very good clotting enzyme for soft-bodied cheese like Serra, but because of its high proteolytic activity it presented some problems in Edam cheese-making. In Roquefort cheese it gave satisfactory results, but with some loss in yield.

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Optimization of Crude Papaya (Carica papaya) Protease in Soft-Unripened Cheese Preparation

Journal of Food Science and Technology Nepal ◽

10.3126/jfstn.v12i12.30139 ◽

2020 ◽

Vol 12 (12) ◽

pp. 1-8

Author(s):

Bunty Maskey ◽

Nabindra Kumar Shrestha

Keyword(s):

Numerical Optimization ◽

Carica Papaya ◽

Enzyme Concentration ◽

The Other ◽

Milk Clotting ◽

Optimization Study ◽

Plant Protease ◽

Milk Clotting Enzyme ◽

Papaya Latex ◽

Milk Clotting Activity

The use of plant protease instead of chymosin for producing cheese has become a trend which is aimed at lacto-vegetarian consumers and religion based ecological markets. In this context, the present investigation was carried out in order to utilize milk clotting enzyme from Papaya (Carica papaya). Numerical optimization study revealed that maximum milk clotting activity was achieved at pH 6.5, temperature 70℃ and enzyme concentration 1 g/1000 ml milk using papaya protease as coagulant. Protein, ash and calcium showed no significant (p>0.05) difference among the cheeses made using different coagulants. However, significantly (p<0.05) higher levels of moisture and ash, and lower levels of fat were observed in the cheese produced by papaya protease compared to that made using rennet. Papaya protease significantly enhanced the spreadability of cheese while the other sensory properties were similar to the control except aftertaste. The results revealed that the papaya latex as crude papaya protease may have potential application for the manufacture of soft-unripened cheese and further could be utilized as a milk coagulant in cheese making.

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Screening and characterization of fungal cultures for a milk-clotting enzyme for use in an oriental style dairy product

Journal of Dairy Research ◽

10.1017/s0022029900031964 ◽

1996 ◽

Vol 63 (3) ◽

pp. 459-466 ◽

Cited By ~ 3

Author(s):

Chin-Wen Lin ◽

Hsiao-Ling Chen

Keyword(s):

Dairy Product ◽

Mucor Racemosus ◽

Milk Clotting ◽

Culture Filtrates ◽

Glutinous Rice ◽

Milk Clotting Enzyme ◽

Pure Cultures ◽

Proteolytic Activities ◽

Rhizopus Javanicus

SummaryThe aim of this work was to develop new oriental style dairy products coagulated with culture filtrates from lao-chao, a traditional fermented rice product. The fermenting fungal cultures screened wereRhizopus javanicus, Rhiz. oryzae, Rhiz. chinensis, Rhiz. oligosporus, Aspergillus oryzae, Mucor racemosusand chiu-yao, a commercial starter. All the inocula liquefied a steamed glutinous rice base and produced culture filtrates with both milk-clotting and proteolytic activities. Of the fungal products tested,Rhiz. javanicuswas the most successful, producing a good yield of culture filtrate with the desired milk-clotting activity, and the resultant yogurt-like product was more acceptable to consumers than any except that from chiu-yao. The firmness was less acceptable, but this could be improved by using mixed pure cultures ofRhiz. javanicusandM. racemosus.

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Influence of different milk-coating enzymes on the process of producing soft cheeses

Food systems ◽

10.21323/2618-9771-2021-4-3-204-212 ◽

2021 ◽

Vol 4 (3) ◽

pp. 204-212

Author(s):

D. S. Myagkonosov ◽

I. T. Smykov ◽

D. V. Abramov ◽

I. N. Delitskaya ◽

V. N. Krayushkina

Keyword(s):

Rhizomucor Miehei ◽

Cost Effective ◽

Milk Clotting ◽

Microbial Origin ◽

Dry Matter Loss ◽

Milk Clotting Enzyme ◽

Negative Effect ◽

Milk Coagulation ◽

Ph Level ◽

Different Origins

The effect of the type of milk clotting enzyme (MCE) on the duration of milk coagulation, parameters of the composition of whey and cheeses, and the output of cheeses in the production of soft cheese such as “Lyubitel'skiy” were investigated. Three brands of MCE of different origins were investigated: Marzyme® (MCE of microbial origin based on Rhizomucor miehei protease), Naturen® (calf rennet) and Chy-max® M (recombinant camel chymosin). It was established that MCEs had different ratios of milk clotting activity (MCA) to total proteolytic activity (PA). It was determined that the MCA/PA ratio, which characterizes the degree of specificity of the MCE action with regard to kappa-casein, in Chy-max M 1000 is ~7 times higher than that of Naturen and ~50 times higher than that of Marzyme. Such differences did not lead to a negative effect when using the Marzyme preparation in the production of soft cheeses. There were no statistically significant differences in the amount of dry matter loss of the curd into the whey, physicochemical parameters and output between the variants of cheeses made with the studied brands of MCE. Shorter duration of milk clotting (16.5 min) was observed with Marzyme than with MCE of Naturen (20.5 min) and Chy-max M (22.5 min). The results of the coagulation duration were explained by the stimulation of the activity of MCE of microbial origin, by the pH level of milk before coagulation (below pH 6.4). It was shown that modern MCEs of microbial origin could be recommended as a cost-effective replacement for more expensive rennet and recombinant chymosins in the production of soft and fresh cheeses.

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