scholarly journals Dynamics of interaction of lactic acid microorganisms in paired consortia aiming at fermentation of vegetable raw materials

2019 ◽  
pp. 96-100
Author(s):  
V. V. Kondratenko ◽  
N. E. Posokina ◽  
J. A. Semenova
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Fermentation Process ◽  
Leuconostoc Mesenteroides ◽  
Chemical Elements ◽  
Raw Materials ◽  
Main Stage ◽  
Main Role ◽  
Rate Of Increase ◽  
Vegetable Raw Materials

Relevance. Cabbage is one of the most popular products, which is mainly fermented with the addition of various vegetables. When fermentation is not only the original nutrients such as vitamin C, amino acids, dietary fibers, etc., but also develop functional microorganisms such as lactic acid bacteria. Fermentation has an important effect on the quality and taste of cabbage, so it is important to study the fermentation process, microbial diversity and changes in nutrients and chemical elements in the fermentation process. L. mesenteroides is considered to be the dominant species on heterofermentative early stages of fermentation. However, there is little information on the diversity of species and strains of Leuconostoc involved in fermentation of sauerkraut. Studies that used traditional biochemical methods to study fermentation of sauerkraut showed that four main types of lactic acid bacteria were involved in the fermentation process: Leuconostoc mesenteroides, Lactobacillus plantarum, Pediococcus pentosaceus and Lactobacillus brevis. Taking into account the importance of two-stage fermentation of vegetable raw materials in order to create optimal conditions for the development of the "main" pool of lactic acid microorganisms at the first stage, it becomes urgent to conduct a complex of studies aimed at reproducing the "natural" process in which the main role is played by bacteria of the genus Leuconostoc mesenteroides at the second stage – monocultures of lactic acid microorganisms and their consortia.Methods. The paper studies the dynamics of the type of interaction of lactic acid microorganisms in paired consortiums on model media pretreated by the culture of the species Leuconostoc mesenteroides, at the main stage of step fermentation of white cabbage of the "Parus" variety.Results. It is established that the sum of the criteria, the consortium "L. mesenteroides \ L. casei + L. plantarum" demonstrates the most pronounced advantage compared with monoculture cultivation of appropriate format of pseudotensorial; despite the pronounced synergy in the cultivation of the consortium "L. mesenteroides \ L. brevis + L. plantarum", the dynamics of the comparison index on the rate of increase in the concentration of microorganisms indicates the need for additional research. 

scholarly journals Influence of consortia of lactic acid microorganisms on the dynamics of active and titratable acidity at the main stage of fermentation of white cabbage

2018 ◽  
Vol 80 (3) ◽  
pp. 140-147
Author(s):  
N. E. Posokina ◽  
E. S. Shishlova ◽  
A. I. Zakharova
Keyword(s):  
Experimental Data ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Fermentation Process ◽  
Raw Materials ◽  
Titratable Acidity ◽  
Starter Cultures ◽  
Main Stage ◽  
White Cabbage ◽  
Direct Fermentation

Fermented cabbage is traditionally produced naturally, where under the action of lactic acid bacteria contained in raw materials, carbohydrates are converted into lactic acid. However, the direct fermentation process does not always lead to an optimal quality product. That is why the use of lactic acid microorganisms is a good alternative to the direct fermentation of cabbage, as with the help of starter cultures the process can be controlled. In this connection, the use of strains of lactic acid microorganisms allows to achieve rapid production of lactic and acetic acids and leads to a rapid decrease in pH, which in turn leads to the suppression of pathogenic microflora, and therefore to the creation of favorable conditions for the fermentation process. The aim of this study was to study the effect of consortia of lactic acid microorganisms on the dynamics of active and titratable acidity in the main stage of fermentation of white cabbage of the variety "Parus". For the study, the prepared modified model medium (MMC) from cabbage was used. In this work, we used lactic acid bacteria Leuconostoc mesenteroides and its consortia: L. mesenteroides + L. casei, L. mesenteroides + L. plantarum, L. mesenteroides + L. brevis, L. mesenteroides + L. casei + L. plantarum, L. mesenteroides + L. plantarum + L. brevis, L. mesenteroides + L. brevis + L. casei. Mathematical processing was carried out according to the obtained experimental data. Analysis of experimental data showed that at the main stage of fermentation the relationship of lactic acid microorganisms in the studied consortia was expressed by synergistic and antagonistic properties. In this case, the best results on the dynamics of growth of active and titratable acidity were obtained in consortiums L. mesenteroides + L. plantarum, L. mesenteroides + L. casei + L. plantarum, L. mesenteroides + L. plantarum + L. brevis.

The Role of Leuconostoc mesenteroides Species of Lactic Acid Bacteria in Fermenting Vegetables

2020 ◽  
pp. 30-36
Author(s):  
NE Posokina ◽  
AI Zakharova
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Fermentation Process ◽  
Raw Materials ◽  
Fermented Food ◽  
Starter Cultures ◽  
Primary Role ◽  
Plant Materials ◽  
Biological Potential ◽  
Fermented Products

Introduction: Fermentation is a biotechnological process of preserving the biological potential of raw materials and transforming them in order to impart new organoleptic properties and to increase nutritional value of the product allowing diversification of daily meals; thus, in some countries fermented products make up a significant part of the human diet. Despite the fact that fermented products are very useful for humans, the fermentation process itself remained rather complicated for reproduction during a long time. Currently, starter cultures are used in industrial production of fermented food products enabling the production of foodstuffs with a guaranteed range of consumer properties. Such species of lactic acid bacteria as Carnobacterium, Enterococcus, Lactobacillus, Lactococcus, Leuconostoc, Oenococcus, Pediococcus, Streptococcus, Tetragenococcus, Vagococcus, and Weissella play the main role in production of fermented food and drinks while L. mesenteroides plays the primary role in starting fermentation of many types of plant materials including cabbage, beet, turnip, cauliflower, green beans, chopped green tomatoes, cucumbers, olives, etc. Objective: To control and manage the industrial fermentation process, it is important to determine the main processes occurring at different stages and the types of lactic acid microorganisms responsible for initiation, continuation and completion of the process. Results: This review shows that, despite the variety of fermentable vegetables, L. mesenteroides species of lactic acid bacteria are of particular importance at the primary heteroenzymatic stage since during this very period the processed raw materials form conditions for inhibiting pathogenic and facultative pathogenic microflora and create optimal environment for subsequent development of targeted microorganisms determining the quality of finished products. Conclusions: When developing food technology, L. mesenteroides species of lactic acid bacteria must be an indispensable component of industrial starter cultures for obtaining final products of consistently high quality.6

scholarly journals Lactic acid bacteria, creating the optimal starting conditions for fermentation of cabbage

2019 ◽  
pp. 80-84
Author(s):  
Nataliya E. Posokina ◽  
Anna I. Zakharova
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Fermentation Process ◽  
Chemical Elements ◽  
Starter Cultures ◽  
Optimal Conditions ◽  
Spontaneous Fermentation ◽  
Microbial Spoilage ◽  
Native Microflora

Relevance Fermentation of vegetables is usually carried out in the traditional way (spontaneous fermentation using native microflora), but the quality of the finished product is difficult to predict. Very often, due to the low initial amount of lactic acid bacteria or their low activity, the result of the process remains unpredictable, which can lead to the loss of a significant amount of product. In the fermentation of vegetables involved several types of facultatively anaerobic lactic acid bacteria. In order to control the fermentation process and make it directed, it is necessary to study which lactic acid bacteria are involved in the fermentation process, the period in which their growth and death, and how it affects the organoleptic properties of the finished product, as well as to study the activity of lactic acid microorganisms in the fermentation process. When fermentation of vegetables are not only the original nutrients such as vitamin C, amino acids, dietary fibers, etc., but also develop functional microorganisms such as lactic acid bacteria. Fermentation has an important effect on the quality and taste, so it is very important to study the fermentation process, microbial diversity and changes in nutrients and chemical elements in the fermentation process. Reducing the rate or preventing microbial spoilage of food is based on four main principles: minimization of product contamination by microorganisms; suppression of growth and reproduction of microorganisms-contaminants; destruction of microorganisms-contaminants; removal of microorganisms-contaminants. Fermentation is based on a combination of the first three principles and is achieved by creating conditions for the growth of specific microorganisms that can give food the desired taste, aroma, texture and appearance. Results This review is devoted to the scientific aspects of vegetable fermentation, including crops that contribute to the creation of optimal conditions for the development of the main pool of lactic acid microorganisms, the production of finished products of high quality and the prevention of microbial spoilage. It is shown that at the first stage of fermentation lactobacilli of the genus L. mesenteroides play a determining role. It is their "work" to create optimal conditions for the development of the target lactic microflora depends on the quality of the finished product. This fact should be taken into account when creating industrial bacterial starter cultures – "starter cultures" for the directed process of fermentation of vegetables.

DESTRUCTION OF CARBOHYDRATES IN THE PROCESS OF DIRECTED FERMENTATION OF PROTEIN CABBAGE WITH A CHANGED CARBOHYDRATE COMPONENT OF THE ENVIRONMENT

2021 ◽  
Author(s):  
В.В. КОНДРАТЕНКО ◽  
Н.Е. ПОСОКИНА ◽  
А.И. ЗАХАРОВА ◽  
М.В. ТРИШКАНЕВА
Keyword(s):  
Lactic Acid ◽  
Food Production ◽  
Raw Materials ◽  
Cost Effective ◽  
Point Of View ◽  
The Body ◽  
Biologically Active ◽  
Main Stage ◽  
White Cabbage ◽  
Vegetable Raw Materials

Польза для здоровья от ферментированных функциональных продуктов выражается непосредственно, через взаимодействие живых микроорганизмов, поступивших в организм вместе с пищевыми продуктами или с биологически активными добавками (пробиотический эффект), или косвенно, в результате потребления микробных метаболитов, синтезируемых в процессе ферментирования (биогенный эффект). Целый ряд исследований посвящен потенциалу процесса ферментирования для создания экономически эффективного производства пищевых продуктов с новыми функциональными и потребительскими свойствами. Присутствие в составе субстрата достаточного количества нутриентов (преимущественно сахаров) играет решающую роль в развитии молочнокислых микроорганизмов в процессе ферментирования. В этой связи были исследованы динамики концентрации глюкозы и фруктозы в результате направленной углеводной корректировки субстрата, приготовленного из белокочанной капусты сорта Парус. Установлено, что оптимальным, с точки зрения деструкции сахаров, является консорциум молочнокислых микроорганизмов видов L. casei и L. plantarum. Экспериментально установлено, что только этот консорциум удовлетворяет условие устойчивого синергизма между входящими в него видами микроорганизмов во время основного этапа ферментирования при условии углеводной корректировки сырья. Процесс углеводной корректировки целесообразно использовать при разработке технологии направленной ферментации овощного сырья. The health benefits of fermented functional products are expressed directly, through the interaction of living microorganisms that have entered the body together with food or biologically active additives (probiotic effect), or indirectly, as a result of consumption of microbial metabolites synthesized during fermentation (biogenic effect). A number of studies are devoted to the potential of the fermentation process to create cost-effective food production with new functional and consumer properties. The presence of a sufficient amount of nutrients (mainly sugars) in the substrate plays a crucial role in the development of lactic acid microorganisms during fermentation. In this regard, the dynamics of glucose and fructose concentrations were investigated as a result of directed carbohydrate adjustment of the substrate prepared from white cabbage of the Parus variety. It has been established that the consortium of lactic acid microorganisms of the species L. casei and L. plantarum is optimal from the point of view of the destruction of sugars. It has been experimentally established that only this consortium satisfies the condition of stable synergy between the species of microorganisms included in it during the main stage of fermentation, subject to carbohydrate adjustment of raw materials. It is advisable to use the process of carbohydrate correction in the development of technology for directed fermentation of vegetable raw materials.

scholarly journals The basics of fermenting white cabbage

2018 ◽  
Vol 80 (2) ◽  
pp. 242-248 ◽  
Author(s):  
E. S. Shishlova ◽  
N. E. Posokina ◽  
O. Yu. Lyalina
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Fermentation Process ◽  
Raw Materials ◽  
Lactobacillus Brevis ◽  
Starter Cultures ◽  
Probiotic Properties ◽  
Acid Fermentation ◽  
Spontaneous Fermentation ◽  
White Cabbage

In this review, the fermentation process (lactic acid fermentation) of white cabbage is completely coveraged. Fermentation is a very complex dynamic process with numerous physical, chemical and microbiological changes affecting quality of the final product. The sequence of lactic acid bacteria development in the fermentation process, which is characterized by the growth and change of pools of various microorganisms, is described. In place of lactic acid microorganisms Leuconostoc mesenteroides comes Lactobacillus brevis, and then propagated Lactobacillus plantarum. The main factors to be taken into account in the fermentation and storage of sauerkraut are given. In order to start the spontaneous fermentation process, it is necessary that the lactic acid bacteria present on the surface of fresh raw materials prevail over the pathogenic microflora. At the same time, the use of starter cultures is a good alternative to natural fermentation, as this ensures the proper flow of the process and the finished product of good quality. The methods of heat treatment, such as pasteurization and sterilization, allowing to extend the shelf life of the finished fermented product. Various types of packaging that are best used for fermented products are also described: plastic bags, glass and metal cans. It is specified what hygienic norms should be observed at production of sauerkraut. It is shown that fermented (fermented) cabbage has probiotic properties that have a beneficial effect on the human body. It is noted that the use of lactic acid microorganisms (starter cultures) in the fermentation process of white cabbage favorably affects the whole process, as it suppresses the development of pathogenic and other undesirable microorganisms on the surface of fresh raw materials and allows to produce a product with improved functional properties.

scholarly journals Study of the dynamics of the development of lactic acid microorganisms in the two-stage process of fermenting cabbage varieties "Parus"

2019 ◽  
pp. 88-93
Author(s):  
Vladimir V. Kondratenko ◽  
Nataliya E. Posokina ◽  
Jeanne A. Semenova ◽  
Vladimir I. Tereshonok
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Fruits And Vegetables ◽  
Leuconostoc Mesenteroides ◽  
Raw Material ◽  
Main Stage ◽  
Fermented Foods ◽  
Acid Fermentation ◽  
Microbiological Methods ◽  
Technological Stage

Relevance. Leuconostoc mesenteroides, Lactobacillus plantarum and Leuconostoc fallaxare the three main lactic acid bacteria developing in the production of sauerkraut. The process of production of fermented foods, as well as their characteristics are determined by the fermenting activity of microorganisms used. Many food products (matured cheeses, sauerkraut, etc.) are canned products: for them, the term is significantly increased compared to the shelf life of the raw material from which these products are made. Their characteristic feature is not only high stability during storage, but also a pronounced flavor bouquet, due to the direct or indirect action of fermenting microorganisms.Material and methods. Microbiological methods of preserving fruits and vegetables are based on the formation of natural preservatives – lactic acid and alcohol, which accumulate as a result of fermentation of sugars by lactic acid bacteria or yeast. The intensity of lactic acid fermentation and the amount of accumulated lactic acid depend on several conditions: the presence of lactic acid bacteria, the content of sugars and other chemicals in the raw material, the addition of spice-aromatic plants, anaerobic conditions, temperature. Due to the accumulation of lactic acid, the development of other microorganisms, and then the lactic acid bacteria themselves, is suspended. The dynamics of development of lactic acid microorganisms and their consortia on the model media, pretreated by the culture of Leuconostoc mesenteroides species, at the main stage of step fermentation of white cabbage of the "Parus"variety are investigated.Results. Dynamics of development of monocultures of L. brevis, L. casei and L. plantarum and their paired consortia at the main technological stage at two stages of fermentation are investigated. It is established that a preliminary fermentation of a model medium with a culture of Leuconostoc mesenteroides generates adverse conditions for the development of L. brevis and L. casei in the monoculture (pseudotensorial) and in consortium with each other, but favorable conditions for the development of L. plantarum in pseudotensorial and in pairwise combinations with L. brevis and L. casei.

Isolation and identification of lactic acid bacteria in Bakasang as fermented microbe starter

2013 ◽  
pp. 48
Author(s):  
J Aquarista Ingratubun ◽  
Frans G Ijong ◽  
Hens Onibala
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Pathogenic Bacteria ◽  
Raw Materials ◽  
Fermented Food ◽  
Plate Count ◽  
Local Market ◽  
Bacillus Sp ◽  
Isolation And Identification ◽  
Total Plate Count

Food fermentation is one of various food processing techniques that has sufficient benefits of nutrition values, and also contains lactic acid bacteria which potentially inhibit pathogenic bacteria, thus prolong shelf life of  products. Bakasang is a traditional fermented food from North Sulawesi since many years ago. Reported research of bakasang previously had described that lactic acid bacteria was the dominant isolates and therefore current research  aimed to isolate and identify the lactic acid bacteria which associated during fermentation day 1 and day 15, respectively. Raw materials used were 5 kg intestine and liver of skipjack brought from local market Bersehati Manado. The intestine and liver of skipjack were washed and smashed and mixed with 10% salt  and 5% rice  from weight of the samples and then filled into bottle to be fermented for 15 days. Every 3 days (1,3,6,9,12,15), the samples were collected and analyzed for total lactic acid bacteria by using Total Plate Count Method on de Mann Rogosa Sharpe Agar after incubation at 37°C for 24 h. The colonies  grown were transferred to Tryptic Soy Broth and followed by streaking them on Tryptic Soy Agar and the free growing colony on agar medium were isolated into slant agar which were used for biochemical test such as Gram’s staining, motility test, catalase test, oksidase test, H2S test, IMVIC test (Indole, Methyl Red, Voges Proskauer, Citrate) and carbohydrate fermentation. The results showed that Lactobacillus sp., Bacillus sp., Eubacterium sp., and Bifidobacterium sp. All these four bacteria were distributed from day 1 to day 15 of the fermentation process© Fermentasi bahan pangan merupakan salah satu dari sekian banyak teknik pengolahan makanan yang mempunyai banyak manfaat dari kualitas gizi, mengandung bakteri asam laktat sehingga menghambat bakteri patogen sehingga daya simpan lebih panjang. Bakasang merupakan makanan fermentasi tradisional masyarakat Sulawesi Utara yang sudah ada sejak lama. Penelitian yang telah dilakukan terhadap bakasang menghasilkan informasi bahwa terdapat bakteri asam laktat pada bakasang sehingga menjadi tujuan untuk mengisolasi dan identifikasi bakteri asam laktat selama proses fermentasi 1-15 hari. Bahan baku bakasang ialah jeroan (usus dan hati) ikan cakalang Katsuwonis pelamis sebanyak 5 kg yang diambil dari pasar Bersehati Manado. Sampel jeroan dibersihkan kemudian dihancurkan, ditambahkan garam 10% dan nasi 5% kemudian difermentasi selama 15 hari dengan mengambil tiap-tiap sampel setiap 1, 3, 6, 9, 12, dan 15 untuk dihitung jumlah bakteri asam laktat dengan menggunakkan metode Total Plate Count pada media de Mann Rogosa Sharpe Agar dan koloni yang tumbuh di tumbuhkan  kembali pada media Tryptic Soy Broth  dan digores kembali pada media Tryptic Soy Agar, koloni yang tumbuh digores pada media slant agar yang selanjutnya diidentifikasi bakteri asam laktat berdasarkan uji biokimia yaitu uji pewarnaan Gram, uji motility, uji katalase, uji oksidase, uji H2S dan uji IMVIC (Indole, MethylRed, Voges Proskauer, Citrate). Hasil menunjukkan bahwa selama proses fermentasi berlangsung terdapat 4 genera bakteri asam laktat sesuai yaitu Lactobacillus sp., Bacillus sp., Eubacterium sp., dan Bifidobacterium sp., ke 4 genera ini tersebar pada fermentasi hari 1 sampai hari ke 15©

scholarly journals Brewers’ spent grain as substrate for dextran biosynthesis by Leuconostoc pseudomesenteroides DSM20193 and Weissella confusa A16

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Prabin Koirala ◽  
Ndegwa Henry Maina ◽  
Hanna Nihtilä ◽  
Kati Katina ◽  
Rossana Coda
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Raw Materials ◽  
Degree Of Polymerization ◽  
Raw Material ◽  
Fermentation Time ◽  
Viscosity Increase ◽  
Spent Grain ◽  
Leuconostoc Pseudomesenteroides ◽  
Weissella Confusa

Abstract Background Lactic acid bacteria can synthesize dextran and oligosaccharides with different functionality, depending on the strain and fermentation conditions. As natural structure-forming agent, dextran has proven useful as food additive, improving the properties of several raw materials with poor technological quality, such as cereal by-products, fiber-and protein-rich matrices, enabling their use in food applications. In this study, we assessed dextran biosynthesis in situ during fermentation of brewers´ spent grain (BSG), the main by-product of beer brewing industry, with Leuconostoc pseudomesenteroides DSM20193 and Weissella confusa A16. The starters performance and the primary metabolites formed during 24 h of fermentation with and without 4% sucrose (w/w) were followed. Results The starters showed similar growth and acidification kinetics, but different sugar utilization, especially in presence of sucrose. Viscosity increase in fermented BSG containing sucrose occurred first after 10 h, and it kept increasing until 24 h concomitantly with dextran formation. Dextran content after 24 h was approximately 1% on the total weight of the BSG. Oligosaccharides with different degree of polymerization were formed together with dextran from 10 to 24 h. Three dextransucrase genes were identified in L. pseudomesenteroides DSM20193, one of which was significantly upregulated and remained active throughout the fermentation time. One dextransucrase gene was identified in W. confusa A16 also showing a typical induction profile, with highest upregulation at 10 h. Conclusions Selected lactic acid bacteria starters produced significant amount of dextran in brewers’ spent grain while forming oligosaccharides with different degree of polymerization. Putative dextransucrase genes identified in the starters showed a typical induction profile. Formation of dextran and oligosaccharides in BSG during lactic acid bacteria fermentation can be tailored to achieve specific technological properties of this raw material, contributing to its reintegration into the food chain.

scholarly journals Hardening Properties of Cheeses by Latilactobacillus curvatus PD1 Isolated from Hardened Cheese-Ddukbokki Rice Cake

2021 ◽  
Vol 9 (5) ◽  
pp. 1044
Author(s):  
Jeong A Kim ◽  
Geun Su Kim ◽  
Se Mi Choi ◽  
Myeong Seon Kim ◽  
Do Young Kwon ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Acid Production ◽  
Lactobacillus Casei ◽  
Leuconostoc Mesenteroides ◽  
The Other ◽  
Rice Cake ◽  
Two Samples ◽  
High Protease Activity

Hardening of cheese is one of major issues that degrade the quality of Home Meal Replacement (HMR) foods containing cheese such as Cheese-ddukbokki rice cake (CD, stir-fried rice cakes with shredded cheese). The quality of cheese, such as pH, proteolytic, and flavor properties, depends on various lactic acid bacteria (LAB) used in cheese fermentation. The hardening of cheese is also caused by LAB. In this study, various LAB strains were isolated from CD samples that showed rapid hardening. The correlation of LAB with the hardening of cheese was investigated. Seven of the CD samples with different manufacturing dates were collected and tested for hardening properties of cheese. Among them, strong-hardening of cheese was confirmed for two samples and weak-hardening was confirmed for one sample. All LAB in two strong-hardening samples and 40% of LAB in one weak-hardening sample were identified as Latilactobacillus curvatus. On the other hand, most LAB in normal cheese samples were identified as Leuconostoc mesenteroides and Lactobacillus casei. We prepared cheese samples in which L. curvatus (LC-CD) and L. mesenteroides (LM-CD) were most dominant, respectively. Each CD made of the prepared cheese was subjected to quality test for 50 days at 10 °C. Hardening of cheese with LC-CD dominant appeared at 30 days. However, hardening of cheese with LM-CD dominant did not appear until 50 days. The pH of the LC-CD was 5.18 ± 0.04 at 30 days, lower than that of LM-CD. The proteolytic activity of LC-CD sample was 2993.67 ± 246.17 units/g, higher than that of LM-CD sample (1421.67 ± 174.5 units/g). These results indicate that high acid production and high protease activity of L. curvatus might have caused hardening of cheese.

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