κ-casein polymorphism effect on technological properties of dried milk

Introduction. Numerous molecular genetic studies have revealed a correlation between the polymorphism of milk protein genes and the technological properties of milk raw materials. DNA analysis, in particular, initiated research into the influence of allelic variants of κ-casein (CSN3) on thermal stability and cheese suitability of milk. This gives relevance to our study that compares the results of genotypic identification of lactating cows by the κ-casein gene, using raw and processed milk samples. Study objects and methods. Our study used raw and reconstituted milk samples from first-calf cows of the black motley breed with the AA and BB genotypes of the κ-casein gene. The samples were analyzed by standardized and generally accepted chemical engineering methods, as well as by capillary electrophoresis and PCR-RFLP analysis. Results and discussion. We compared the results of tests on thermal stability and cheese suitability of raw and reconstituted milk samples from cows with the AA and BB genotypes of the κ-casein gene. We tried out an integrated approach to monitoring milk raw materials based on the most relevant technological criteria and correlating the data with the associated CSN3 gene identification parameters. The PCR-RFLP analysis revealed reproducible results for both raw and dried milk samples in relation to the genotypical identification by the A- and B- allelic variants of the CSN3 gene. The tests showed higher thermal stability in the reconstituted milk from the BB genotype cow and better cheese suitability in the AA genotype sample. Conclusion. We developed a system for evaluating milk raw materials based on the most important technological parameters in combination with their genotypic characteristics. Our research procedure can unify the accumulation of experimental data and contribute to the formation of bioinformatics algorithms. This approach can be used in mathematical modeling of criteria to evaluate the compliance of the technological properties of milk with the recommended indicators.

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New determination method for the ratio of the relative proportions of ϰ-casein alleles in milk powder

Food Processing Techniques and Technology ◽

10.21603/2074-9414-2020-3-525-535 ◽

2020 ◽

Vol 50 (3) ◽

pp. 525-535

Author(s):

Khamid Gilmanov ◽

Vladislav Semipyatniy ◽

Alana Bigaeva ◽

Ramil Vafin ◽

Svetlana Turovskaya

Keyword(s):

Dairy Products ◽

Raw Materials ◽

Milk Powder ◽

Rflp Analysis ◽

Raw Milk ◽

Raw Material ◽

Technological Properties ◽

Additional Indication ◽

Casein Gene ◽

Pcr Rflp

Introduction. High-quality dairy products depend on the properties of raw materials. Milk is the basic raw material for all dairy products. Its quality is subject to requirements of biological safety, and its physicochemical and technological properties comply with strict indicators. ϰ-casein gene is a basic milk protein. Its allelic variants affect milk productivity, as well as its composition and technological properties, e.g. cheeseability and heat resistance. The research objective was to develop a new method for determining the ratio of the relative proportions of selection-significant kappa-casein gene alleles in bulk cow’s milk powder by means of molecular genetics and bioinformation system. The method can help to test raw materials for further processing rationality. Study objects and methods. The research included a genetic analysis of samples of bulk whole milk powder, bulk powdered skimmed milk, and bulk raw milk. The developed method involved DNA extraction, combined PCR-RFLP technique, electrophoretic results, and analysis detection of information data using new mathematical algorithms and software. Results and discussion. The ratio of the relative proportions of the kappa-casein gene alleles in milk powder was determined using primer sets JK5 and JK3, as well as restriction endonuclease HinfI for PCR-RFLP analysis. The experiment showed satisfactory reproducibility and interpretation of the obtained data. The program provided a ratio calculation of the kappa-casein gene alleles relative proportions in the studied milk powder samples, expressed as a percentage of the A allele proportion with additional indication of the absolute and relative errors. It also placed the information block of the generated numerical indicators into the percentage scale. Conclusion. The developed methodology was implemented by a set of laboratory procedures. The new specialized program “Calculation of the relative proportions ratio of ϰ-casein alleles in bulk milk” is on open access. It provides a correct and prompt data interpretation, generated during the analysis of dry milk raw materials of bulk origin.

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Evaluation of the Effect of κ-casein Gene Polymorphism in Milk Powder on the Technological Properties of Acid-Induced Milk Gels

Food Processing Techniques and Technology ◽

10.21603/2074-9414-2021-1-53-66 ◽

2021 ◽

Vol 51 (1) ◽

pp. 53-66

Author(s):

Alexandr Kruchinin ◽

Svetlana Turovskaya ◽

Elena Illarionova ◽

Alana Bigaeva

Keyword(s):

Mechanical Properties ◽

Gene Polymorphism ◽

Mass Fraction ◽

Raw Materials ◽

Milk Powder ◽

Storage Conditions ◽

Technological Properties ◽

Casein Gene ◽

Bulk Milk ◽

Temperature Storage

Introduction. Yoghurt has become one of the most popular acid-induced dairy products in the world. Consumers see yoghurt not only as a tasty, protein-rich, and calcium-fortified dessert, but also as a product that improves intestinal microflora and prevents obesity, metabolic syndrome, type II diabetes, and cardiovascular diseases. The stability of the structure and consistency of yoghurt directly depend on the composition and properties of raw materials, including genetic and technological factors and compliance with temperature storage conditions. Yoghurt formulations include various dairy raw materials, e.g. milk powder. The research objective was to assess the effect of κ-casein gene polymorphism in milk powder on the technological properties of acid-induced milk gels when simulating different temperature storage conditions. Study objects and methods. The research featured yoghurt samples prepared from milk powder of CSN3 gene (AA and BB). Model systems of yoghurt were prepared from dry bulk milk, mixed by mass fraction of protein in the ratio of AA2:BB2 as 75:25, 50:50, and 25:75%, respectively. The experiment involved standard methods, optical methods, dynamic viscometry, and PCR-RFLP. Results and discussion. As the mass fraction of BB dry bulk milk increased, the structural and mechanical properties, dimensional stability, and surface tension increased, too. As the storage temperature fell from 4 ± 2 to 12 ± 2°C, the structural and mechanical properties, dimensional stability, surface tension, and moisture-holding ability decreased while maintaining the previously established dependencies. The CSN3 gene polymorphism proved to have no effect on the curd tension after fermentation. Significant differences between the allelic variants AA and BB became obvious only after complete cooling and structuring of the product. Conclusion. The obtained experimental results and the analysis of related publications suggested an indirect effect of the κ-casein gene polymorphism on the structural and mechanical properties, associated with a genetic effect on the average diameter of casein micelles in the original milk and the resulting biochemical and isothermal processes. The research made it possible to assess the effect of the CSN3 gene polymorphism on the technological properties of dry milk during its processing into fermented milk products.

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Genetic polymorphisms of cytochrome P450 enzymes: CYP2C9, CYP2C19, CYP2D6, CYP3A4, and CYP3A5 in the Croatian population

Drug Metabolism and Personalized Therapy ◽

10.1515/dmpt-2016-0024 ◽

2017 ◽

Vol 32 (1) ◽

Cited By ~ 5

Author(s):

Lana Ganoci ◽

Tamara Božina ◽

Nikica Mirošević Skvrce ◽

Mila Lovrić ◽

Petar Mas ◽

...

Keyword(s):

Rflp Analysis ◽

The Other ◽

Published Data ◽

Cytochrome P450 Enzymes ◽

Chain Reaction ◽

Allelic Variants ◽

Mediterranean Populations ◽

Pcr Rflp ◽

Polymerase Chain ◽

European Populations

AbstractBackground:Data on the frequency of pharmacogenetic polymorphisms in the Croatian population are limited. We determined and analyzed frequencies for the most importantMethods:2637 subjects were included. Genotyping was performed by real-time polymerase chain reaction (PCR) using TaqMan® DME or TaqMan® SNP Genotyping Assays, and by PCR, and PCR-RFLP analysis.Results:ForConclusions:The frequency of the CYP allelic variants, genotypes, and predicted phenotypes in the Croatian population is in accordance with the other European populations, between the values of published data for Middle European and Mediterranean populations.

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A PCR‐RFLP Test to Detect Allelic Variants of the Bovine Kappa‐Casein Gene

Animal Biotechnology ◽

10.1081/abio-120020180 ◽

2003 ◽

Vol 14 (1) ◽

pp. 1-5 ◽

Cited By ~ 11

Author(s):

L. A. Soria ◽

G. M. Iglesias ◽

M. J. Huguet ◽

S. L. Mirande

Keyword(s):

Allelic Variants ◽

Casein Gene ◽

Pcr Rflp

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The Use of Genetic Marking in Breeding Work with Breed Herds

Vestnik APK Verhnevolzh`ia ◽

10.35694/yarcx.2020.50.2.005 ◽

2020 ◽

Author(s):

Ю.А. Михайлова ◽

Р.В. Тамарова

Keyword(s):

Dairy Products ◽

Cottage Cheese ◽

Frequency Of Occurrence ◽

Technological Properties ◽

Dna Testing ◽

Allelic Variants ◽

Casein Gene ◽

Breeding Work ◽

Yield And Quality

Исследования проведены по актуальной проблеме повышения белковомолочности и улучшения технологических свойств коровьего молока и качества белковомолочных продуктов. Цель исследований – проследить наследование аллелей гена каппа-казеина дочерьми от быков-отцов с АВ и ВВ генотипами каппа-казеина, оценить белковомолочность коров, технологические свойства молока, выход и качество творога из молока коров ярославской породы с разными генотипами каппа-казеина. Основные исследования проводились в племрепродукторе по ярославской породе Ярославской области с 2009 года, а в одном из племзаводов региона изучались технологические свойства молока коров, генотипированных по каппа-казеину, с 2012 года. В специализированной лаборатории ДНК-технологий ФГБНУ ВНИИплем были исследованы образцы крови по современной методике ДНК-тестирования каппа-казеина. Решающую роль играют быки-производители в наследовании признака белковомолочности. Было прогенотипировано 49 коров-дочерей от 11 быков-отцов преимущественно чистопородных ярославских животных. Установлено наследование с учётом генотипов каппа-казеина быков-отцов и дочернего потомства. Наследование аллельных вариантов гена каппа-казеина дочерьми от быков различное. Выявлены два препотентных быка-производителя, передающих с наибольшей частотой встречаемости В-аллельные варианты каппа-казеина дочерям (частота встречаемости 0,5). Получены данные выхода и качества белковомолочной продукции (творога) у В-аллельного варианта каппа-казеина на 14–29% выше, чем из молока коров с генотипом АА. Рентабельность производства творога из молока коров, имеющих в генотипе аллель В каппа-казеина, выше на 22–46%. Выявленные данные позволяют рекомендовать шире использовать ДНК-тестирование по генотипу каппа-казеина при селекции крупного рогатого скота по экономически ценным признакам – по содержанию белка и технологическим свойствам молока. Researches have been conducted on the recent problem of increasing protein and milking capacity and improving the technological properties of cow's milk and the quality of protein and dairy products. The goal of the research was to trace the inheritance of alleles of the kappa casein gene by daughters from bulls-fathers with AB and BB genotypes of kappa casein, to evaluate the protein and milking capacity of cows, the technological properties of milk, the yield and quality of cottage cheese from milk of cows of the Yaroslavl breed with different genotypes of kappa casein. The main researches have been conducted in a pedigree breeding unit for the Yaroslavl breed of the Yaroslavl region since 2009 and in one of the stud farms of the region the technological properties of the milk of cows genotyped by kappa casein have been studied from 2012. In a specialized DNA technology laboratory of the FSBSI All-Russian Scientific Research Institute of Breeding work blood samples were examined according to the modern DNA testing technique of kappa casein. The decisive role is played by the servicing bulls in the inheritance of the sign of protein and milking capacity. 49 daughter cows from 11 father bulls of predominantly pure-bred Yaroslavl animals were genotyped. Inheritance has been established taking into account the kappa-casein genotypes of bull fathers and daughter offspring. The inheritance of the allelic variants of the kappa-casein gene by the daughters from the bulls is different. Two prepotent bull sires were identified transmitting B-allelic variants of kappa-casein to daughters with the highest frequency of occurrence (frequency of occurrence 0.5). The data on the yield and quality of protein and dairy products (cottage cheese) were obtained in the B-allelic variant of kappa-casein 14–29% higher than from milk of cows with genotype AA. The profitability of the production of cottage cheese from the milk of cows with the kappa casein allele B in the genotype is 22–46% higher. The identified data make it possible to recommend the wider use of DNA testing according to the genotype of kappa casein in the selection of cattle according to economically valuable features – according to the protein content and technological properties of milk.

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REAL-TIME PCR TECHNOLOGY FOR CATTLE GENOTYPING BY A AND B KAPPA-CASEIN GENE ALLELES

SERIES CHEMISTRY AND TECHNOLOGY ◽

10.32014/2021.2518-1491.16 ◽

2021 ◽

pp. 126-132

Author(s):

R. R. Vafin ◽

Kh. Kh. Gilmanov

Keyword(s):

Real Time ◽

Fluorescence Detection ◽

Real Time Pcr ◽

Bos Taurus ◽

Rflp Analysis ◽

Casein Gene ◽

Analysis Technique ◽

Pcr Rflp ◽

Allele Specific ◽

Fluorescence Quencher

The main goal of the study was to develop and test an effective technology for cattle genotyping by the CSN3 gene based on real-time PCR with hybridization-fluorescence detection. There was developed a method for real-time PCR for cattle genotyping by A and B alleles of the CSN3 gene in the format of hybridization-fluorescence detection, involving the use of two 5/-fluorescence-labeled forward allele-specific primers, one reverse common primer, and one anti-primer labeled with a fluorescence quencher at the 3/-end of the oligonucleotide. As a result of practical studies aimed at testing the developed method, we obtained the technical result provided by the proposed technology, expressed in the effective identification of the desired genotypes due to correct interpretation of these curves of increasing fluorescence intensity, the results reliability of which was also confirmed by the well-known PCR-RFLP analysis technique for Bos taurus genotyping for similar allelic variants of the kappa-casein gene.

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Calpastatin (CAST) Gene Polymorphism in Tsigai and Merinoland Sheep Breeds Under Conditions of the Republic of Crimea

BIO Web of Conferences ◽

10.1051/bioconf/20213606002 ◽

2021 ◽

Vol 36 ◽

pp. 06002

Author(s):

Veronika Uppe ◽

Tatiana Kuevda ◽

Denis Zubochenko ◽

Elena Usmanova ◽

Pavel Ostapchuk

Keyword(s):

Gene Polymorphism ◽

Research Methods ◽

Rflp Analysis ◽

Primary Objective ◽

Ongoing Study ◽

Allelic Variants ◽

Study Research ◽

Pcr Rflp ◽

The Republic ◽

Cast Gene

The article presents the results of studying the Calpastatin (CAST) gene polymorphism in Tsigai (n = 23) and Merinoland sheep breed (n = 13). The purpose of the research. The possibility of using PCR-RFLP analysis to detect the Calpastatin (CAST) gene polymorphism in Tsigai and Merinoland sheep was the primary objective of this ongoing study. Research methods. Calpastatin (CAST) gene polymorphism was analyzed by PCR-RFLP analysis using the MspI endonuclease restriction. Results. We revealed the diversity of genotypes and allelic variants of the Calpastatin (CAST) gene in Tsigai and Merinoland Breeds. It was found that M allele of the Calpastatin locus is the most common. Frequencies of MM, MN and NN genotypes were found to be 74, 4 and 22 % in Tsigai breed. Among the representatives of Merinoland sheep, the frequencies of MM and MN genotypes were 92 % and 8%, respectively. No animals with NN genotypes were found.

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