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ТЕХНИЧЕСКИЕ НАУКИ
RESEARCH OF THE EFFECTS OF ENZYME PREPARATION WITH COLLAGENASE ACTIVITY ON THE PROTEIN FRACTION OF COOKED STRUCTURAL SHORT SHELF-LIFE SAUSAGES
Indzhelieva D.
Indzhelieva Diana — PhD., Chief Assistant, PROF. DR ASEN ZLATAROV UNIVERSITY - COLLEGE OF TOURISM, BURGAS, REPUBLIC OF BULGARIA
Abstract: the purpose of this paper is to investigate the influence of bacterial enzyme preparation with collagenase activity on the protein fraction status in short shelf-life structural boiled and smoked sausages made of raw meat rich in connective tissue. Studies have have been carried out with an enzyme preparation having a collagenase activity derived from Clostridium histolyticum, producing SIGMA ALDRICH, a branch of Merck. Collagenase enzyme preparation is added at the meat cutting in the form of aqueous solution in the following concentrations: 10 000 50 000 and 90 000 U/kg filling mass.
The results obtained show that the enzyme preparation contributes to increasing the degree of protein hydrolysis in the raw meat. This is expressed in:
1. Increase the amount ofpeptides and free amino acids in test samples.
2. Increase the amount of free amino nitrogen.
3. The use of a higher activity of the collagenase enzyme preparation gives rise to a high degree of hydrolysis.
Keywords: meat products, enzyme preparations.
ИССЛЕДОВАНИЕ ВЛИЯНИЯ ФЕРМЕНТНОГО ПРЕПАРАТА С АКТИВНОСТЬЮ КОЛЛАГЕНАЗЫ НА БЕЛКОВЫЕ ФРАКЦИИ СТРУКТУРНЫХ ВАРЕНЫХ КОЛБАС Инджелиева Д. Т.
Инджелиева Диана Тодорова - кандидат технических наук, главный ассистент, Университет им. профессора д-ра Ас. Златарова, Колледж по туризму, г. Бургас, Республика Болгария
Аннотация: целью данной работы является исследование влияния бактериального препарата фермента с активностью коллагеназы на состояние белковой фракции в структуре скоропортящихся варёно-копченых колбас, в производстве которых использовалось сырое мясо, богатое соединительной тканью. Исследования были проведены с препаратом фермента, имеющего активность коллагеназы, полученной из Clostridium histolyticum, производитель Sigma Aldrich, филиал Merck. Препарат коллагеназы-фермент добавляют при резке мяса в виде водного раствора в следующих концентрациях: 10 000 50 000 и 90 000 U/кг масса наполнения.
Полученные результаты показывают, что препарат фермента способствует повышению степени гидролиза белков в сыром мясе. Это выражается в:
1. Увеличении количества пептидов и свободных аминокислот в опытных образцах.
2. Увеличении количества свободного аминного азота.
3. Использовании более высокой активности препарата коллагеназы фермента, что приводит к высокой степени гидролиза.
Ключевые слова: мясные продукты, ферментные препараты. Introduction
Boiled short shelf-life sausages take a leading position in the meat processing sector worldwide. Consumers' interest in these sausages is associated with their nutritional and energy value, as well as with a large assortment diversity. From manufacturers' point of view, cooked short shelf-life sausages have a number of advantages that allow for their manufacturing as a great variety of products. In the modern manufacturing of boiled short shelf-life sausages, an increasing trend towards the use of raw meat with low functional properties has been observed. The use of such raw material that is rich in
connective tissue, if not pre-leaned, leads to deteriorated quality of the finished products. The use of new methods in the meat processing technology aiming at acceleration of the biochemical processes in the raw meat is related to the improvement of their technological and organoleptic properties [7,8]. These methods enable the intensification of the technological processes that allows for shortening the duration of the manufacturing cycle. It also creates an opportunity to utilize the raw meat with reduced technological properties by its utilization in sausages and canned products.
There are studies that have established the hydrolytic activity of the widely used enzymes of plant origin in the meat processing industry, such as papain, bromelain and ficain, and the bacterial collagenase preparation "Sigma VII" [5]. The maximum activity of "Sigma VII" enzyme preparation occurs at approximately 50-60°C, and the enzyme preparation becomes inactive at temperature above 60°C. The insufficient leaning effect of the enzyme on beef is explained by the authors with the low proteolytic activity of the enzyme preparation.
There are also research data published on the ability of three microbial collagenase preparations derived from Clostridium histolyticum, Vibrio B-30, and Achomobacter jophagus to decompose the bovine muscle tissue [3]. Investigators have added collagenase preparations (with and without amylase added) to shredded beef. They have established a significant increase in the solubility of collagen as compared to the samples, in which there was no enzyme preparation added.
In the last decade, an increasing number of studies have covered the possibilities for enzymatic processing of low-functional raw meat [2, 4, 6]. In this regard, the objective hereof is to determine the effects of the bacterial enzyme preparation with collagenase activity on the protein fraction status in cooked structural short shelf-life sausages made of raw meat rich in connective tissue. Materials and methods
Studies have been carried out with an enzyme preparation with collagenase activity derived from Clostridium histolyticum, manufactured by SIGMA ALDRICH, Merck's branch, with the following characteristics:
appearance - powder
colour - beige
activity according to Mandl - U / mg optimal temperature of activity - 37-40°C pH optimum - 6,3 - 7,4
Boiled and smoked structural short shelf-life sausages have been used to determine the effects of the collagenase enzyme preparation on the protein fraction status of cooked short shelf-life sausages.
Studies establishing the effects of the enzyme preparation in structural cooked short shelf-life sausages have been conducted based on the following recipe (kg/100 kg of filling mass): Pork 30
Bacon 15
Bovine or swine hearts 12
Meat from pigs heads with cheeks and skin 40 Wheat flour 3
Salt 0.200
Sodium nitrite 0.005
Pentasodium tripolyphosphate 0.200
Black (white) pepper 0.300
Paprika (hot pepper) 0.300
Garlic 0.200
Sausages are prepared by cutting the pork meat and hearts to pieces, mincing them in a Wolf-machine with 3-4 mm diameter of the openings and placing them in a cutter machine for fine mincing. During the cutting process, salt, nitrite, polyphosphate and finely crushed ice or ice water amounting to 25-30% of the meat mass weight are added. Add the spices and flour, and then the pork head meat and bacon, preliminary minced in the Wolf-machine with 10 mm diameter of the openings. Cutting continues for 4-6 min until the pork head meat and bacon are cut to 4-5 mm particles.
Add the enzyme preparation during the cutting for preparation of the filling mass. This ensures a good possibility for uniform distribution of the preparation between the meat particles.
Collagenase enzyme preparation is added in the form of aqueous solution in the following concentrations: 10 000, 50 000 and 90 000 U/kg filling mass. The ready filling mass is filled into natural sausage casings and is taken for thermal treatment.
Meanwhile, control samples are produced with no enzyme preparation added thereto. To establish the effects of the enzyme preparation on the protein fraction status of the finished sausages, the changes in the amino nitrogen, degree of hydrolysis, amount of free amino acids and
polypeptides need to be tracked. The degree of hydrolysis of the protein fraction is calculated by referring the amount of free a-amino nitrogen to the total quantity of nitrogen in the sample, determined by the method of Kjeldahl. The amount of a-amino nitrogen is determined by Sorensen formol titration method and the amount of free amino acids and polypeptides is determined according to АОАС, 1995 [1].
Statistical processing of the data obtained is performed by „STATPLUS 2009" software.
Results and discussion
Results obtained from the studies aiming at establishing the appropriateness of use of collagenase enzyme additives during the preparation of structural short shelf-life sausages show that the use of collagenase results in increased degree of protein hydrolysis and increase in the amount of free amino nitrogen as compared to sausages untreated with enzyme (Fig. 1 and 2). This is due to the proteolysis of meat proteins induced by the impact of the enzyme preparation added. The use of the enzyme preparation is accompanied by formation and accumulation of free amino nitrogen in the sausages.
As it can be seen, by increasing the concentration of the enzyme preparation used, the amount of the free amino nitrogen increases as a result of the flowing process of proteolysis in a greater degree.
mg% 40
35 30 25 20 15 10 5 0
Fig. 1. Amount of amino nitrogen (mg °%) in Fig. 2. Degree of hydrolysis (%>) in structural
structural sausages according to the activity of sausages according to the activity of the enzyme
the enzyme preparation preparation
% 1,8
1,6 1,4 1,2 1 0,8 0,6 0,4 0,2 0
U/kg U/kg U/kg U/kg U/kg U/kg
Fig. 3. Amount ofpolypeptides in structural Fig. 4. Degree of free amino acids (% of dry
sausages according to the activity of the enzyme substance) in structural sausages according preparation (% of dry substance) to the activity of the enzyme
% 1,8 1,6 1,4 1,2
0,8 0,6
0,4
0,2
0 100005000090000 U/kg U/kg U/kg
0 10000 50000 90000 U/kg U/kg U/kg
1
0
The results relating to the effects of the collagenase preparation on the content of free amino acids and polypeptides in structural sausages are shown on Fig. 3 and 4. As it can be seen, by increasing the concentration of the enzyme the amount of free amino acids is also increased. There is an increase in the amount of free amino acids in the sausages prepared with the use of enzyme preparation.
From the data shown in Fig. 3 it is obvious that there is a significant increase in the amount of polypeptides in the sausages prepared with the enzyme preparation, as compares with the control sample (without enzyme). By increasing the activity of the enzyme used, the increase of polypeptides is insignificant. Conclusions
The results obtained and the analysis thereof give us reason to assume the appropriateness of use of bacterial enzyme preparation with collagenase activity in manufacturing of structural short shelf-life boiled and smoked sausages rich in connective tissue.
It has been found that enzyme preparation contributes to increasing the degree of protein hydrolysis in raw meat used for manufacturing of short shelf-life boiled and smoked sausages. This is expressed in the following:
1. Increase of polypeptide and free amino acids amount in the test samples.
2. Increase of free amino nitrogen amount.
3. The use of higher activity of collagenase enzyme preparation results in a higher degree of hydrolysis.
References
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4. FDA. Carbohydrase and protease enzyme preparations derived from Bacillus subtilis or Bacillus amyloliquefaciens; Affirmation of GRAS status as direct food ingredients. 21 CFR. Part 184. Federal Register, 23 April, 1999. Vol. 64. № 78. P. 19887-19896.
5. Foegeding E. A. and Larick D. K. Tenderization of beef with bacterial collagenase // Meat Sci., 1986. № 18. P. 201-214.
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7. Miller M. F., Carr M. A., Ramsey C. B., Crockett K. L. and Hoover L. C. Consumer thresholds for establishing the value of beef tenderness // J. Anim. Sci., 2001. № 79. P. 3062-3068.
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