CA2162181A1 - Accelerated maturation of cheddar cheese by the addition of live and heat-shocked lactobacilli and neutrase - Google Patents
Accelerated maturation of cheddar cheese by the addition of live and heat-shocked lactobacilli and neutraseInfo
- Publication number
- CA2162181A1 CA2162181A1 CA 2162181 CA2162181A CA2162181A1 CA 2162181 A1 CA2162181 A1 CA 2162181A1 CA 2162181 CA2162181 CA 2162181 CA 2162181 A CA2162181 A CA 2162181A CA 2162181 A1 CA2162181 A1 CA 2162181A1
- Authority
- CA
- Canada
- Prior art keywords
- cheese
- neutrase
- heat
- months
- casei
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C19/00—Cheese; Cheese preparations; Making thereof
- A23C19/06—Treating cheese curd after whey separation; Products obtained thereby
- A23C19/063—Addition of, or treatment with, enzymes or cell-free extracts of microorganisms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C19/00—Cheese; Cheese preparations; Making thereof
- A23C19/06—Treating cheese curd after whey separation; Products obtained thereby
- A23C19/061—Addition of, or treatment with, microorganisms
- A23C19/062—Addition of, or treatment with, microorganisms using only lactic acid bacteria, e.g. pediococcus, leconostoc or bifidus sp., or propionic acid bacteria; Treatment with non-specified acidifying bacterial cultures
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Microbiology (AREA)
- Dairy Products (AREA)
Abstract
Live and heat-shocked Lactobacillus casei subsp casei (L2A) cultures, supplemented with the proteolytic enzyme Neutrase, have been developed by means of biotechnology, which accelerate Cheddar cheese ripening by up to 60% in flavor intensity, as compared to control cheese. The present invention studied Cheddar cheese evolution in detail over a 9-month ripening period by evaluating microbiological, physico-chemical, sensory and rheological parameters.
Description
- - ~,AZ~621~ 2 This novel integrated process comprises the addition of (t) live ~acfo~ac~llus casei subsp casel L2A to control the undesirable micro!lora, ~2) heat-shocked cells of the same culture at a level of 1% (v/v), and (3) Neutrase at a level of 1.0 x 10 5 Au/g of ci~eese.
LA~::TOBAC~LLI CELLS AND EXTRACTS
FOR ~CCELERATED CHE~SE RIPENING
The production of matured Chedd~r cheese involves considerable co~s for the cheese industry, mainly due to a slow microbial process which incurs higll costs for refri~eration and warehousing.
Furlhermore, impending legisiation for mandatory pasteurization to destroy pathogens (e.g.
List~ria) will lower the content of useful lactic acid bacteria (LAB), increasin~ the time necessary to produce maturecl cheese.
~ Cheese flavor development is a dynamic process which represents a finely orchestrated series of successive and concomitant biochemical events over a period of time, leading to products with highly desirable aromas and flavors. None are ch~racterized sufficiently to permit dup~ication of their complele flavor by mixtures of pure compounds lhus using indirect methods to speed up -cheese ripening (1~.
Elevated temperatures during cheese ripening, added enzymes, modilied starter and slurry method have been commonly used, but ~he most widely employed method for accelerated cheese ripening is the addition of extraneous enzyrmes to the cheese (2).
~. CA2162181 Although some cheap commercial food Qrade enzymes are available for the above purpose, almost all have their limitations, especially regarding the control of their action on milk components, resulting in a rheologically poor final product, and often with bitter flavor.
Lactococcus Jactis enzymes were introduced to the market as the product "Accelerase" by Imperial Biotechnology Ltd (London), but most starter lactococci are unable to multiply in cheese and contain less active peptidases ~3), and esterases (4~ than Lactobacillus strains (5).
The present novel process involves the addition of live and heat-shocked cells of Lactobacillus casei subsp-casei L2A, combined with the enzyme Neutrase and elevated ripening temperature.
Ti~e Lactobacillus strain selected was typical of the microfiora of good quality Cheddar cheese, and shows good growth and ensured control of undesirable microflora (6, 7). This strain is known for its rich peptidases (4), which are responsible for adval1ced hydrolysis of milk casein, and in order to develop desirable flavors without quality defects.
DESCRI-?TION OF THE INVENTION
Fresh raw milk was pasteurized (73~C, 16 s) immediately before cheesemaking, and cheeses were made in pilot cheese vats containing 225 L of milk. The starter culture, a mixture of Lactococc~ls cremorisand Leucor~ostoccremoris(Agropur, Granby, Quebec, Canada) was added to the milk (1.5% viv) after three transfers for 18 'nr at 20~C in 12% (w/v) reconstituted skim milk at a level of 2% (vlv) inoculum for the acidification of all vats. Lactobaci/Jus casei-subsp-casei i~A
grown in 12% (w/v) reconstituted skim milk (30"C, 48 hr) was added to the milk at 0.01% (v/v) inoculum.
When the milk acidity had increased by 4~ Dornic (0.04% Lactic Acid), rennet (50% bovine rennet + 50% porcine pepsin; Chr. i-iansens Lab, Inc., Milwaukee, Wl) was added at a level of 0.02%
(v/v). The miik was set at 30~C for 30 min, the curd was cut, cooked at 38~C, and the whey drained off. After cheddaring at 38~C, milling, and salting (2%, w/w), following overnight pressing of the curd (approx. 10 kg), all cheeses were vacuum packed in plastic bags (4 mil, Winpak Co., Winnipeg, Canada), and ripened at 4~C for one week, foilowed by 2 months at 13~C, and 7 months at 7~C.
Heat-shocked lactobacilli were prepared by adjusting the pH of the culture to 6.5 using 1 M
NaOH, and by pumping through a stainless stell coil immersed in a water bath (67~C) to achieve a 22-s contact time. Mortality rate was 94.5% but proteolytic activity, measured by Hide Powder Azure (HPA) method (8) remained unchanged. Duplicate cheeses were made on separate days in four identical vats and the effects of the various concentrations of bacterial additives compared.
After milling, the curd from each vat was divided in two and different concentrations of Neutrase added.
Control cheese, which constituted the first of three groups, is a standard cheese without the additives. Live and heat-shocked cu!tyres were added to the second group of cheeses, while various concentrations of Neutrase besides bacterial additives were added to the third group. The experimental design is shown in Table 1.
Cheese samples taken after ~, 1, 2, 3, 4 weeks and 2, 4, 6, 9 months were analyzed for lactic acid bacteria (LAB) and lactobacilli counts. LAB were counted on Lactobacilli MRS agar (Difco), following incubation at 30~C for 48 h under anaerobic conditions (BBL Gaspak system).
Lactobacilli numbers were determined with Rogosa agar (Difco). Each cheese sample (25 g) was sliced and added to 225 ml of 2.0% citrated water ahd homogenized for 5 min in a Stomacher (Lab Blender, Model 400, A.J. Seward Lab., London, UK). Dilutions were plated on Lactobacilli MRS and Rogosa Agars.
Samples of cheeses were taken for duplicate determination of fat, protein, moisture, salt and pH
by APHA (9) methods. Physico-chemical ratios were calculated as:
% M/NFS = (Moisture/Non-fat solids (100-fat)) X 100, % F/S = (FaVSolids (100-moisture)) X 100, % SIM = (Salt/Moisture) X 100.
Rheological parameters \Ivere determined by double compression tests using the Instron Universal Testing Machine (Model 1101, Instron Corp., Canton, U.S.A.). The 500-kg cell was fixed to the crosshead, adjusted to 5 cm/min, and chart speed was 5 cmimin.
~A21 621 81 The compression unit and a stainless steel cylinder (Diameter: 3.5 cm) caused 80% deformation of cylindrical cheese samples (diarneter: 1.25 cm; height: 1.0 cm) held at room temperature l hr before testing. Three parameters, fracturability (kPa), firn~ness (kPa), and cohesiveness (~,h), were studied with ten replicates of each cheese. Cheeses coded with three-digit numbers were presented to a panel of three expert graders from Agriculture Canada at 1, 2, 4, 6 and 9 months of ripening. Two blocks of cheese were graded for flavor and texture. Flavor intensity was graded on a scale of 1 to 1 O, while flavor and texture defects were evaluated on a scale of 1 to 100. The rninimum points of grade 1, 2, 3 Canada Cheddar were 2 92, 91 to 87, and < 86, respectively.
Analysis of variance ~F test) was performed on microbial counts, proximate analysis (moisture, fat, protein, salt), physico-chernical (pH, fracturability, firmness, cohesiveness), and flavor intensity (%~ by sensory evaluation.
The variance homogeneity was verified by the Bartlett test, and Duncan's multiple range test used to determine significant differences (p c 0.05) among treatments (10).
The results of the experimental design for cheesemaking showed that the proxirnate analysis (moisture, fat, proteinl salt) of the one month-old cheeses was comparable for all cheeses, within the normal range for Cheddar cheese, except for protein content (Table 2).
The protein content of the experimental cheeses was significantly higher than the control cheese throughout the ripeniny period (Fig. 1). This increase was likely due to the presence of the bacterial and enzyme additives. l~he pH va!ues of experimental cheeses were lower than the control cheese (Fig. 2), mainly due to the production of lactic acid by live lactobacilli. Where added, Neutrase liberated neutral peptides that partly neutralized the lactic acid.
Significant difference in LAB and lactobacilii counts were observed between the control and experimental cheeses up to 2 months, except for the beginning of rnaturation (Figs. 3, 4).
Cheeses produced with added cultures of lactobacilli had LAB counts of 7.0 to 8.5 log within 1 week, while that of the control was 5.7 log/g of cheese. Lactobacilli counts showed a similar growth pattern to the LAB, but the corresponding control counts were 4.0 log, while the - ~A21621 81 experimental cheeses had an average cour,t of 6.0-6.7 log/g. Cheeses supplemented with either 1.0 to 2.0% of heat-shocked ceils showed comparable counts because of the rapid growth ot residual live lleat-shocked ceils. Although certain strains of Lactobacillus casei contributed to increased acidity to the point where the cheeses were graded as acidic and bitter-tasting (11), the addition of lactobacilli cultures, heat-shocked cells and Neutrase did not affect the cheese quality. All cheeses were classified as first class and many of them would qualify as premium class (Table 3). The pH values were between 5.01 and 5.13. pl I values above 5.2 or below 4.85 are considered undesirable (12). Low pH is known to influence cheese texture by decreasing the fracturation force (13, 14), but the effect of pH was not clearly demonstrated (l~ig. 5), because of the more pronounced effect of Neutrase on the weakening of the casein network. Fracturability, firmness and cohesiveness values in Figs 5, 6 and 7 showed a decreasing pattern over the whole maturation period, with relative stabilization during the last 3 months. These changes are due to the degradation of ~5,-casein, and agree with the results of others (8, 13, 15). Cheese texture continued to soften during the last 3 months, but was counterbalanced by the rise in pH, thus leading to a final stabilization of rheological parameters.
Cheeses supplemented with Neutrase at a concentration of 4 X 10 5 Au/g of cheese were more fracturable and less cohesive than those with Neutrase at 2.0 or l.0 X 105 Au/g, thereby confirming the results of others (8, 15). The effect of heat-shocked cells on rheological parameters compared with Neutrase was not significant, as its activity was mainly directed towards the degradation of small peptides as well as casein. We confirmed this previously by a good correlation between the nitrogen soluble in phosphotungstic acid (PTA-sol N) and the amount of heat-shocked cells added ( 16). The interpretation of rheological parameters is sometimes rendered difficult, as evidenced in firmness (Fig. 6). This large variability associated with rheological measurements is often caused by the non-homogeneity of cheese (17, 18, l9).
Statistical analysis of rheological parameters showed that the addition of Neutrase appears to be the most effective treatment, unless this causes sensory deterioration.
Flavor intensity scores given by expert graders permitted calculation of % increase in flavor during maturation. A good correlation was found among % increase in (l) flavor intensity, (2) heat-shocked cells (HS) and (3) Neutrase concentration.
~ .
~ ~, 'A 2 l ~ 1 8 ~ ; ~7~
The order of efficiency in acceleraUng ripening at 6 months was: HS (2.0%) i Neutrase B > HS
(~.G%) ~ Neutrase C ~ HS (1%) + Neutrase A, B, C. Aithough % increases varied from 100% to 50%, HS (2.0%) plus Neutrase C showed a significant.y greater increase in flavor intensity as cornpared to the other l-eal~nenls throughout maturation, except at 6 months. These results compared favorably with the reduction of 30 to 50% in ripening time claimed by many authors (9, 21, 22, 23, 24).
The quali~y of control cheese and cheeses supplemented with live (LL) and heat-shocked ~HS) cells remained excellent (class 1) throughout maturation. However, total cheese quality (flavor &
texture) was strongly influenced by pitte~ess development in cheeses with added Neutrase.
Cheeses which received live + ~IS celis and Neutrase showed a good quali~ (class ll) after 6 and 9 months. However, bittemess developed significantly with increased PJeutrase concentration.
Tex~ure defects were not directly associa.ed with intensity of treatment but parUy with ptl of the cheese.
Based on total quality score and % increase in flavs~r, this novei process recommends the addition of 1.0C~o HS cells, along with Neutrase (1.0 X 105 Aulg of cheese). To control the undesirable microflora, addition of live Lactobacillus casei-sut~sp-caseJ (~A) cells is also suggested at a concentration not higher than 4.0 (Log,0 CFU/ml of milk) to maintain the desired pH.
This novel process does not appear to be expensive, ~iven the ~ow c~ncentrations of additives necessary, and the simple apparatus required for the heat-shock treatment. When we combine the effect of elevated storage temperature, as shown in the presentstudy, ~his process represents an important technolo~y for accelerating Cheddar cheese ripening.
., . , . ,, . ~
CA~ l 6~
REFERENCES CITED
1. Olson, N. 1990. FEMS Microbiology Reviews 87:131-148.
LA~::TOBAC~LLI CELLS AND EXTRACTS
FOR ~CCELERATED CHE~SE RIPENING
The production of matured Chedd~r cheese involves considerable co~s for the cheese industry, mainly due to a slow microbial process which incurs higll costs for refri~eration and warehousing.
Furlhermore, impending legisiation for mandatory pasteurization to destroy pathogens (e.g.
List~ria) will lower the content of useful lactic acid bacteria (LAB), increasin~ the time necessary to produce maturecl cheese.
~ Cheese flavor development is a dynamic process which represents a finely orchestrated series of successive and concomitant biochemical events over a period of time, leading to products with highly desirable aromas and flavors. None are ch~racterized sufficiently to permit dup~ication of their complele flavor by mixtures of pure compounds lhus using indirect methods to speed up -cheese ripening (1~.
Elevated temperatures during cheese ripening, added enzymes, modilied starter and slurry method have been commonly used, but ~he most widely employed method for accelerated cheese ripening is the addition of extraneous enzyrmes to the cheese (2).
~. CA2162181 Although some cheap commercial food Qrade enzymes are available for the above purpose, almost all have their limitations, especially regarding the control of their action on milk components, resulting in a rheologically poor final product, and often with bitter flavor.
Lactococcus Jactis enzymes were introduced to the market as the product "Accelerase" by Imperial Biotechnology Ltd (London), but most starter lactococci are unable to multiply in cheese and contain less active peptidases ~3), and esterases (4~ than Lactobacillus strains (5).
The present novel process involves the addition of live and heat-shocked cells of Lactobacillus casei subsp-casei L2A, combined with the enzyme Neutrase and elevated ripening temperature.
Ti~e Lactobacillus strain selected was typical of the microfiora of good quality Cheddar cheese, and shows good growth and ensured control of undesirable microflora (6, 7). This strain is known for its rich peptidases (4), which are responsible for adval1ced hydrolysis of milk casein, and in order to develop desirable flavors without quality defects.
DESCRI-?TION OF THE INVENTION
Fresh raw milk was pasteurized (73~C, 16 s) immediately before cheesemaking, and cheeses were made in pilot cheese vats containing 225 L of milk. The starter culture, a mixture of Lactococc~ls cremorisand Leucor~ostoccremoris(Agropur, Granby, Quebec, Canada) was added to the milk (1.5% viv) after three transfers for 18 'nr at 20~C in 12% (w/v) reconstituted skim milk at a level of 2% (vlv) inoculum for the acidification of all vats. Lactobaci/Jus casei-subsp-casei i~A
grown in 12% (w/v) reconstituted skim milk (30"C, 48 hr) was added to the milk at 0.01% (v/v) inoculum.
When the milk acidity had increased by 4~ Dornic (0.04% Lactic Acid), rennet (50% bovine rennet + 50% porcine pepsin; Chr. i-iansens Lab, Inc., Milwaukee, Wl) was added at a level of 0.02%
(v/v). The miik was set at 30~C for 30 min, the curd was cut, cooked at 38~C, and the whey drained off. After cheddaring at 38~C, milling, and salting (2%, w/w), following overnight pressing of the curd (approx. 10 kg), all cheeses were vacuum packed in plastic bags (4 mil, Winpak Co., Winnipeg, Canada), and ripened at 4~C for one week, foilowed by 2 months at 13~C, and 7 months at 7~C.
Heat-shocked lactobacilli were prepared by adjusting the pH of the culture to 6.5 using 1 M
NaOH, and by pumping through a stainless stell coil immersed in a water bath (67~C) to achieve a 22-s contact time. Mortality rate was 94.5% but proteolytic activity, measured by Hide Powder Azure (HPA) method (8) remained unchanged. Duplicate cheeses were made on separate days in four identical vats and the effects of the various concentrations of bacterial additives compared.
After milling, the curd from each vat was divided in two and different concentrations of Neutrase added.
Control cheese, which constituted the first of three groups, is a standard cheese without the additives. Live and heat-shocked cu!tyres were added to the second group of cheeses, while various concentrations of Neutrase besides bacterial additives were added to the third group. The experimental design is shown in Table 1.
Cheese samples taken after ~, 1, 2, 3, 4 weeks and 2, 4, 6, 9 months were analyzed for lactic acid bacteria (LAB) and lactobacilli counts. LAB were counted on Lactobacilli MRS agar (Difco), following incubation at 30~C for 48 h under anaerobic conditions (BBL Gaspak system).
Lactobacilli numbers were determined with Rogosa agar (Difco). Each cheese sample (25 g) was sliced and added to 225 ml of 2.0% citrated water ahd homogenized for 5 min in a Stomacher (Lab Blender, Model 400, A.J. Seward Lab., London, UK). Dilutions were plated on Lactobacilli MRS and Rogosa Agars.
Samples of cheeses were taken for duplicate determination of fat, protein, moisture, salt and pH
by APHA (9) methods. Physico-chemical ratios were calculated as:
% M/NFS = (Moisture/Non-fat solids (100-fat)) X 100, % F/S = (FaVSolids (100-moisture)) X 100, % SIM = (Salt/Moisture) X 100.
Rheological parameters \Ivere determined by double compression tests using the Instron Universal Testing Machine (Model 1101, Instron Corp., Canton, U.S.A.). The 500-kg cell was fixed to the crosshead, adjusted to 5 cm/min, and chart speed was 5 cmimin.
~A21 621 81 The compression unit and a stainless steel cylinder (Diameter: 3.5 cm) caused 80% deformation of cylindrical cheese samples (diarneter: 1.25 cm; height: 1.0 cm) held at room temperature l hr before testing. Three parameters, fracturability (kPa), firn~ness (kPa), and cohesiveness (~,h), were studied with ten replicates of each cheese. Cheeses coded with three-digit numbers were presented to a panel of three expert graders from Agriculture Canada at 1, 2, 4, 6 and 9 months of ripening. Two blocks of cheese were graded for flavor and texture. Flavor intensity was graded on a scale of 1 to 1 O, while flavor and texture defects were evaluated on a scale of 1 to 100. The rninimum points of grade 1, 2, 3 Canada Cheddar were 2 92, 91 to 87, and < 86, respectively.
Analysis of variance ~F test) was performed on microbial counts, proximate analysis (moisture, fat, protein, salt), physico-chernical (pH, fracturability, firmness, cohesiveness), and flavor intensity (%~ by sensory evaluation.
The variance homogeneity was verified by the Bartlett test, and Duncan's multiple range test used to determine significant differences (p c 0.05) among treatments (10).
The results of the experimental design for cheesemaking showed that the proxirnate analysis (moisture, fat, proteinl salt) of the one month-old cheeses was comparable for all cheeses, within the normal range for Cheddar cheese, except for protein content (Table 2).
The protein content of the experimental cheeses was significantly higher than the control cheese throughout the ripeniny period (Fig. 1). This increase was likely due to the presence of the bacterial and enzyme additives. l~he pH va!ues of experimental cheeses were lower than the control cheese (Fig. 2), mainly due to the production of lactic acid by live lactobacilli. Where added, Neutrase liberated neutral peptides that partly neutralized the lactic acid.
Significant difference in LAB and lactobacilii counts were observed between the control and experimental cheeses up to 2 months, except for the beginning of rnaturation (Figs. 3, 4).
Cheeses produced with added cultures of lactobacilli had LAB counts of 7.0 to 8.5 log within 1 week, while that of the control was 5.7 log/g of cheese. Lactobacilli counts showed a similar growth pattern to the LAB, but the corresponding control counts were 4.0 log, while the - ~A21621 81 experimental cheeses had an average cour,t of 6.0-6.7 log/g. Cheeses supplemented with either 1.0 to 2.0% of heat-shocked ceils showed comparable counts because of the rapid growth ot residual live lleat-shocked ceils. Although certain strains of Lactobacillus casei contributed to increased acidity to the point where the cheeses were graded as acidic and bitter-tasting (11), the addition of lactobacilli cultures, heat-shocked cells and Neutrase did not affect the cheese quality. All cheeses were classified as first class and many of them would qualify as premium class (Table 3). The pH values were between 5.01 and 5.13. pl I values above 5.2 or below 4.85 are considered undesirable (12). Low pH is known to influence cheese texture by decreasing the fracturation force (13, 14), but the effect of pH was not clearly demonstrated (l~ig. 5), because of the more pronounced effect of Neutrase on the weakening of the casein network. Fracturability, firmness and cohesiveness values in Figs 5, 6 and 7 showed a decreasing pattern over the whole maturation period, with relative stabilization during the last 3 months. These changes are due to the degradation of ~5,-casein, and agree with the results of others (8, 13, 15). Cheese texture continued to soften during the last 3 months, but was counterbalanced by the rise in pH, thus leading to a final stabilization of rheological parameters.
Cheeses supplemented with Neutrase at a concentration of 4 X 10 5 Au/g of cheese were more fracturable and less cohesive than those with Neutrase at 2.0 or l.0 X 105 Au/g, thereby confirming the results of others (8, 15). The effect of heat-shocked cells on rheological parameters compared with Neutrase was not significant, as its activity was mainly directed towards the degradation of small peptides as well as casein. We confirmed this previously by a good correlation between the nitrogen soluble in phosphotungstic acid (PTA-sol N) and the amount of heat-shocked cells added ( 16). The interpretation of rheological parameters is sometimes rendered difficult, as evidenced in firmness (Fig. 6). This large variability associated with rheological measurements is often caused by the non-homogeneity of cheese (17, 18, l9).
Statistical analysis of rheological parameters showed that the addition of Neutrase appears to be the most effective treatment, unless this causes sensory deterioration.
Flavor intensity scores given by expert graders permitted calculation of % increase in flavor during maturation. A good correlation was found among % increase in (l) flavor intensity, (2) heat-shocked cells (HS) and (3) Neutrase concentration.
~ .
~ ~, 'A 2 l ~ 1 8 ~ ; ~7~
The order of efficiency in acceleraUng ripening at 6 months was: HS (2.0%) i Neutrase B > HS
(~.G%) ~ Neutrase C ~ HS (1%) + Neutrase A, B, C. Aithough % increases varied from 100% to 50%, HS (2.0%) plus Neutrase C showed a significant.y greater increase in flavor intensity as cornpared to the other l-eal~nenls throughout maturation, except at 6 months. These results compared favorably with the reduction of 30 to 50% in ripening time claimed by many authors (9, 21, 22, 23, 24).
The quali~y of control cheese and cheeses supplemented with live (LL) and heat-shocked ~HS) cells remained excellent (class 1) throughout maturation. However, total cheese quality (flavor &
texture) was strongly influenced by pitte~ess development in cheeses with added Neutrase.
Cheeses which received live + ~IS celis and Neutrase showed a good quali~ (class ll) after 6 and 9 months. However, bittemess developed significantly with increased PJeutrase concentration.
Tex~ure defects were not directly associa.ed with intensity of treatment but parUy with ptl of the cheese.
Based on total quality score and % increase in flavs~r, this novei process recommends the addition of 1.0C~o HS cells, along with Neutrase (1.0 X 105 Aulg of cheese). To control the undesirable microflora, addition of live Lactobacillus casei-sut~sp-caseJ (~A) cells is also suggested at a concentration not higher than 4.0 (Log,0 CFU/ml of milk) to maintain the desired pH.
This novel process does not appear to be expensive, ~iven the ~ow c~ncentrations of additives necessary, and the simple apparatus required for the heat-shock treatment. When we combine the effect of elevated storage temperature, as shown in the presentstudy, ~his process represents an important technolo~y for accelerating Cheddar cheese ripening.
., . , . ,, . ~
CA~ l 6~
REFERENCES CITED
1. Olson, N. 1990. FEMS Microbiology Reviews 87:131-148.
2. Law, B.A. Flavor developrnent in cheese. In: advances in the microbiology andbiochemistry of cheese and fermented milk (Davies, F.L. and B.A. Law, eds), pp 209-228, Academic Press, London, 1984.
3. Arora, G. and Lee, ~.H. 1990. J. Dairy Sci. 73:274-279.
4. Lee. S.Y. and Lee, B.H. 1989. J. Foods Sci. 54:119-122 & 126.
5. Lee, B.H., Haché, S. and Simard, R.E. 1986. J. Ind. Microbiol. 1:209-217.
6. Laleye, L.C., Simard, R.E., Le~,~B.H. and Holley, R.A. J. Dairy Sci. 72:3134-3142.
7. Lee, B.H., Laleye, L.C., Simard, R.E., Munsch, M.H. and ~oliey, R.A. 1990. J. Food Sci.
55:391 -3g7.
55:391 -3g7.
8. Law, B.A. and Wigmore, A.S. 1g82. J. Dairy Res. 49:137.
9. APHA. Standard Methods For the Examination of Dairy Products (Richardson, G.H. ed), 15th edition. American Publich Health Association, Washington, D.C., U.S.A. 1985.
10. Steel, R.G.D. and Torrie, J.H. Principles and Procedure of Statistics. A Biometrical approach, 2nd edition, McGraw-Hill Book Cornpany, New-York, N.Y. 1980.
11. Lee, B.H., Laleye, L.C., Simard, R.E., Holley, R.;~., Emmons, D.B. and Giroux, R.N.1989.
J. Food Sci. 55:386-390.
J. Food Sci. 55:386-390.
12. Gilles, J. and Lawrence, R.C. 1973. New Zeal. J. Dairy Sci. & Technol. 8:148-13. Cream~r, L.K. and Olson, N.F. 1982. J. Food Sci. 47:631-14. Creamer, L.K., J. Gilles and Lawrencel R.C.1988. New Zeal. J. Dairy Sci. Technol.23:23-15. Fedrick, l.A. and Dulley, T.R. 1984. New Zeal. J.Dairy Sci. Technol. 19:141-16. El Abboudi, M., Pandian, S., Trépanier, G., Simard, R.E. and Lee, B.H.1992. J. Food Sci.
17. Lee, C.H., lmoto, E.M. and Rha, C. 1978. J. Food Sci. 43:1600.
18. Walstra, P. and Van Vliet, T. 1982. Int. Dairy Fed. Bull. Doc. 153:22.
19. Trépanier, G., Simard, R.E. and Lee, B.H. 1991. J. Food Sci. 53:696-700.
20. El-Soda, M., Desmazeaud, M.J., Aboudonia, S. and Badvan, A. 19~2. Michwissenschaft 37:325-21. ~I-Soda, M., Ezzart, N., El Deeb, S., Mashly, R.l. and Moustapha, F.1986. Le Lait 66:177-22. Abdelbaky, A.A., El-Neshawy, A.A., Rabie, A.M. and Ashour, M.M. lg86. Food Chem.
21 :301-- l,A2162181 o, 23. Fedrick, I.A., Cromie, S.J., Dulley, J.R. and Gilks, J.E. 1986. New Zeal. i. Dairy Sci.
Technol. 21:191.
. ~
T~ 4:1'ERCENTAGElNCl~EASElN FLAVORINTENSITY' TREATMENT ,~ MATURAliON TlME(months) LIVELACTO
+ o n12.5 50.() 3.1 i~SLACTO1.(1~
+ NF,l)'rRASE
LIVELACTO A 33012.5 62.5 (1.0 + B 50.0 375 5().0 (1.0 HSLACTO1.n',~ C 33.0 ~37 5 fi2.: fi.25 LIVELACTO B 17.025.1) lUU.O 0.0 HSLACTO2.0i~ C X3.05U() ~7.5 12.5 a: ~ vori!lt(~ it~ p~rim~llt~ cc X 100~
~ Q~vorin~ y~f ~ntrol c~
qb Table 3: SEr~SiOKY EVALUATION OF CHEESE L)UKING MATURATI- N
TREATMENT MATlJKA l lC)N TIME ~months) ~ ! 2 ¢ fi 9 T S' TEXTb T S TEXT T S T EX1 T S TEXT T S TEXT
CONTROL 92.() 1.0 92.(1 1.0 92() 1.() 92.0 1.0 93.0 0.0 LIVE LACTO
+ Y2 5 ().0 92.0 0.0 '33 () ().() 93.0 ().0 92.2 0.3 HS LAC'TO l.O~o + NEUTRASE
LIVE LACTO A 92.0 0.5 92.4 0.5 92.5 0.5 92.0 0.5 91.0 0.5 + B 92.() 0.5 92.5 1.0, 92 5 ().() 9().9 1.5 91.0 0.5 HS LACTO 1 0~ C 92.() 0.5 92.5 1.0 92.0 1.() 89.5 1.5 88.0 1.0 Ll~'E l AC-rO B 92.() 0.~s 91 0 05 ~2.5 U.0 8'3.0 1.5 90.0 0.0 HS LACTO 2.()~. C 92 0 1.0 9().0 0.5 '3~.() 0.5 88 0 2.0 89.0 0.0 a: Total quality (flavor and texture) score; mcan val-le (n = I to 2); scal~ of I to 100: Class 1: 2 92 {'I.lSS 11: 91 to ~7 Cl~-iss ]11: < 86 h: Texture score; mean value (n = I to 2), scale of 0 to 3 (normal texture to w~k or acid texture).
~.
r ~-Tablc 2: I'ROXIMATE Al\IAl.YSIS OF ONE-MON1 1~-OL,D CHEESES' T}~EATMENT MOISIlJRE FAT l'~OTEIN SALT pH
% c~o % ~/u CONTKOL 34.43b 35.0()c29.6&1 1.37f 5.27 LIVE LACTO
+ 34.34b 33.fi2c25.42e 1.55f 5.01h HS LACrO 1.()'7u + NEIJTI~ASE
LrVE LACTO A 33.76b 34.~3c 25.9()e 1.32f 5.14i t 1~ 33.t~b 34.75c25.fifi~ 1.50f 5.13i HS LACrO 10% C 34.12b 34.62c 25.35e 1.55f 5.13i LIVE LACTO B 34.31b 35.25c 25.54e 1.53f 5.10j +
HS LACTO 2.0% C 33.82b 34.25c 25.fi4c 1.33f 5.13j ;3: mcan valuc (n = 1 to 2). MCIII5 in tlle SdnlC COlllllln IlOt follow~1 by a COlnrllOII letler are significantly diffcrcnt (P < 0.05~.
i (e~-g~-) a~ou ql~ pue .~ Jo ~e~e (~cj~j a~ F~ l F: r. ~ u:sr~ u!u c I U! C o ;o S~ ase.~aU! :e ~noqe ~u~q o~ ~-a.r ~a~ a~i'~,J.t ,0 ~uno:ue =nV ~ x ~, n uosu~ :~
OGt Y ~ !!'U 30 au ~ u/~ o~ auru!nllo~ ;~inili~ e 3~ aurlG~) ~'Z i iaSe~-i;)Se~ sr~ tqo;ae-;, 2U i;eS 2~ -,V 5.0l X GG ~ se 3na~ 1 zaul~auua~T ~6C~O Z + ove- SH ~=
du~;auua~ ~6l0C ~ ove~
t 3S~iJ !~.
9u~ ec ~/!lV S.~l X ooz~ ase:;ni~N l a z~uli~uuay ~G~ Z + o ~e I sH I
2u~;auuad ~6100 + o~el I
2U~ J v/~l~v o- x G~ ;r~
- 2ui;-u::aY ~ .01 ~ . ci_e-l ST~) . ac~v I C3~ 0HS-l -H
~u-;~ uua~.: ~6 i û G . o- e~ ~_ 21 i~-~s ~ s ~ ~ .~e n.~
~ z8uilauu.~ o;ai--l SH l 2 u~.u. aY -~ .G ~ e~
7~uiuec ~ OG I V asEI~ni~
7aUna'i'J~'Ci '~ I . o'ai'~; SH I IllT~V~0~V1 3Nl D
2UilaUUa~Y ~Q~ + c3ai~, 1 2uilauuay ~GO I o: i-. SH .(o~ae~i sHi ~ 'V~0s--~1 a3~1X'HS-'V~H
Zaullauu i~T ~b IG ~G o~ (olar l~ T:~VSO I ~Vl 3.~i ~V N ~ V N ;e~ pJrp~r~S lO~TLl~i'OZ) oo 3 'VlSNO"~ VY~ H ~NO~ _ S IVZ)lld3~NoiLTaav 3,Ullcav .NriT~l~_iS3a 3/~T.. 'iaiTV lN3W1~3~1 N '!S~(l lVlt\,'3Wii3dX3 :i alCel 5)
21 :301-- l,A2162181 o, 23. Fedrick, I.A., Cromie, S.J., Dulley, J.R. and Gilks, J.E. 1986. New Zeal. i. Dairy Sci.
Technol. 21:191.
. ~
T~ 4:1'ERCENTAGElNCl~EASElN FLAVORINTENSITY' TREATMENT ,~ MATURAliON TlME(months) LIVELACTO
+ o n12.5 50.() 3.1 i~SLACTO1.(1~
+ NF,l)'rRASE
LIVELACTO A 33012.5 62.5 (1.0 + B 50.0 375 5().0 (1.0 HSLACTO1.n',~ C 33.0 ~37 5 fi2.: fi.25 LIVELACTO B 17.025.1) lUU.O 0.0 HSLACTO2.0i~ C X3.05U() ~7.5 12.5 a: ~ vori!lt(~ it~ p~rim~llt~ cc X 100~
~ Q~vorin~ y~f ~ntrol c~
qb Table 3: SEr~SiOKY EVALUATION OF CHEESE L)UKING MATURATI- N
TREATMENT MATlJKA l lC)N TIME ~months) ~ ! 2 ¢ fi 9 T S' TEXTb T S TEXT T S T EX1 T S TEXT T S TEXT
CONTROL 92.() 1.0 92.(1 1.0 92() 1.() 92.0 1.0 93.0 0.0 LIVE LACTO
+ Y2 5 ().0 92.0 0.0 '33 () ().() 93.0 ().0 92.2 0.3 HS LAC'TO l.O~o + NEUTRASE
LIVE LACTO A 92.0 0.5 92.4 0.5 92.5 0.5 92.0 0.5 91.0 0.5 + B 92.() 0.5 92.5 1.0, 92 5 ().() 9().9 1.5 91.0 0.5 HS LACTO 1 0~ C 92.() 0.5 92.5 1.0 92.0 1.() 89.5 1.5 88.0 1.0 Ll~'E l AC-rO B 92.() 0.~s 91 0 05 ~2.5 U.0 8'3.0 1.5 90.0 0.0 HS LACTO 2.()~. C 92 0 1.0 9().0 0.5 '3~.() 0.5 88 0 2.0 89.0 0.0 a: Total quality (flavor and texture) score; mcan val-le (n = I to 2); scal~ of I to 100: Class 1: 2 92 {'I.lSS 11: 91 to ~7 Cl~-iss ]11: < 86 h: Texture score; mean value (n = I to 2), scale of 0 to 3 (normal texture to w~k or acid texture).
~.
r ~-Tablc 2: I'ROXIMATE Al\IAl.YSIS OF ONE-MON1 1~-OL,D CHEESES' T}~EATMENT MOISIlJRE FAT l'~OTEIN SALT pH
% c~o % ~/u CONTKOL 34.43b 35.0()c29.6&1 1.37f 5.27 LIVE LACTO
+ 34.34b 33.fi2c25.42e 1.55f 5.01h HS LACrO 1.()'7u + NEIJTI~ASE
LrVE LACTO A 33.76b 34.~3c 25.9()e 1.32f 5.14i t 1~ 33.t~b 34.75c25.fifi~ 1.50f 5.13i HS LACrO 10% C 34.12b 34.62c 25.35e 1.55f 5.13i LIVE LACTO B 34.31b 35.25c 25.54e 1.53f 5.10j +
HS LACTO 2.0% C 33.82b 34.25c 25.fi4c 1.33f 5.13j ;3: mcan valuc (n = 1 to 2). MCIII5 in tlle SdnlC COlllllln IlOt follow~1 by a COlnrllOII letler are significantly diffcrcnt (P < 0.05~.
i (e~-g~-) a~ou ql~ pue .~ Jo ~e~e (~cj~j a~ F~ l F: r. ~ u:sr~ u!u c I U! C o ;o S~ ase.~aU! :e ~noqe ~u~q o~ ~-a.r ~a~ a~i'~,J.t ,0 ~uno:ue =nV ~ x ~, n uosu~ :~
OGt Y ~ !!'U 30 au ~ u/~ o~ auru!nllo~ ;~inili~ e 3~ aurlG~) ~'Z i iaSe~-i;)Se~ sr~ tqo;ae-;, 2U i;eS 2~ -,V 5.0l X GG ~ se 3na~ 1 zaul~auua~T ~6C~O Z + ove- SH ~=
du~;auua~ ~6l0C ~ ove~
t 3S~iJ !~.
9u~ ec ~/!lV S.~l X ooz~ ase:;ni~N l a z~uli~uuay ~G~ Z + o ~e I sH I
2u~;auuad ~6100 + o~el I
2U~ J v/~l~v o- x G~ ;r~
- 2ui;-u::aY ~ .01 ~ . ci_e-l ST~) . ac~v I C3~ 0HS-l -H
~u-;~ uua~.: ~6 i û G . o- e~ ~_ 21 i~-~s ~ s ~ ~ .~e n.~
~ z8uilauu.~ o;ai--l SH l 2 u~.u. aY -~ .G ~ e~
7~uiuec ~ OG I V asEI~ni~
7aUna'i'J~'Ci '~ I . o'ai'~; SH I IllT~V~0~V1 3Nl D
2UilaUUa~Y ~Q~ + c3ai~, 1 2uilauuay ~GO I o: i-. SH .(o~ae~i sHi ~ 'V~0s--~1 a3~1X'HS-'V~H
Zaullauu i~T ~b IG ~G o~ (olar l~ T:~VSO I ~Vl 3.~i ~V N ~ V N ;e~ pJrp~r~S lO~TLl~i'OZ) oo 3 'VlSNO"~ VY~ H ~NO~ _ S IVZ)lld3~NoiLTaav 3,Ullcav .NriT~l~_iS3a 3/~T.. 'iaiTV lN3W1~3~1 N '!S~(l lVlt\,'3Wii3dX3 :i alCel 5)
Claims (6)
1. A process for preparing lactococci starler and lactobacilli adjunct cultures in pilot plantvats wherein 225 L of fresh raw milk was pasteurized (73°C, 16s), inoculated with a mixted culture (1:1 ratio) of Lactococcus cremoris and Leuconostoc cremoris at a level of 1.5%
(v/v) after three transfers grown tor 18 hr at 20°C in 12% (w/v) reconstituted skim milk, and that Lactobacillus casei-subsp-casei L2A grown in 12% (w/v) reconstituted skim milk (30°C, 48 hr) was added to the milk at a level of 0.01% (v/v), before renneting.
(v/v) after three transfers grown tor 18 hr at 20°C in 12% (w/v) reconstituted skim milk, and that Lactobacillus casei-subsp-casei L2A grown in 12% (w/v) reconstituted skim milk (30°C, 48 hr) was added to the milk at a level of 0.01% (v/v), before renneting.
2. A process for making Cheddar cheese in pilot plant scale according to Claim 1 in which rennet was added at a level of 0.02% (v/v) when milk acidity had increased by 4° Domic (0.04% Lactic Acid), the curd was cut, cooked, and cheddared at 38°C, followed by milling, salting (2%, w/w), packing and ripening at 4°C for one week, and then 2 months at 13°C
and 7 months at 7°C.
and 7 months at 7°C.
3. A process for preparing heat-schocked Lactobacillus casei-subsp-casei L2A cells on a laboratory scale, wherein the culture was adjusted to pH 6.5 using 1 M NaOH and passed through a stainless steel coil immersed in water bath (67°C) for 22 s to achieve mortality rate of 94.5% without destroying proteolytic enzyme activity, and the heat-shocked cells were added to cheeses prepared according to claim 2 at 1.0% or 2.0% (v/v) levels.
4. A process for making Cheddar cheese on a pilot plant scale according to claim 3, wherein different concentrations of the proteolytic enzyme Neutrase, at (1) 1.0 X 10-5 (Arson Unit), (2) 2.0 X 10-5 Au, and (3) 4.0 X 10-5 Au/g of cheese were added at salting stage.
5. A combined process for making Cheddar cheese by adding (1) 0.01% (v/v) of live Lactobacillus casei-subsp-casei L2A cells of claim 1, (2) 1.0% or 2.0% (v/v) levels of heat-shocked cells of claim 3, (3) Neutrase at 1.0 x 10-5 Au and/or 2.0 X 10-5 Au/g of cheese as in claim 4, and by maturing those cheeses at 4°C for one week, followed by 2 months at 13°C, and 7 months at 7°C, as in claim 2.
6. A process for reducing the Cheddar cheese ripening period by up to 60% on a laboratory scale, pilot scale or commercial scale according to any of claims 1 to 6 inclusive.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA 2162181 CA2162181A1 (en) | 1995-11-23 | 1995-11-23 | Accelerated maturation of cheddar cheese by the addition of live and heat-shocked lactobacilli and neutrase |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA 2162181 CA2162181A1 (en) | 1995-11-23 | 1995-11-23 | Accelerated maturation of cheddar cheese by the addition of live and heat-shocked lactobacilli and neutrase |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2162181A1 true CA2162181A1 (en) | 1997-05-24 |
Family
ID=4156908
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA 2162181 Abandoned CA2162181A1 (en) | 1995-11-23 | 1995-11-23 | Accelerated maturation of cheddar cheese by the addition of live and heat-shocked lactobacilli and neutrase |
Country Status (1)
| Country | Link |
|---|---|
| CA (1) | CA2162181A1 (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101795573A (en) * | 2007-09-27 | 2010-08-04 | 明治乳业株式会社 | Method of producing natural cheese |
| EP3056088A3 (en) * | 2016-04-13 | 2016-11-02 | DSM IP Assets B.V. | Cheese ripening |
-
1995
- 1995-11-23 CA CA 2162181 patent/CA2162181A1/en not_active Abandoned
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101795573A (en) * | 2007-09-27 | 2010-08-04 | 明治乳业株式会社 | Method of producing natural cheese |
| EP3056088A3 (en) * | 2016-04-13 | 2016-11-02 | DSM IP Assets B.V. | Cheese ripening |
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| Date | Code | Title | Description |
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Effective date: 19981015 |