Development of the New Method of the Melted Cheese Products Without Salt-melters Using Cryomechanolysis

The aim of the work is elaboration of the principally new method of deep processing of rennet cheeses to the melting using the complex action of freezing and cryomechanolysis on the raw material that gives a possibility to destruct the hardly soluble biopolymers and to transform them into soluble form.The principally new method of the deep processing of rennet cheeses for receiving the melt cheese products without salts-smelters was elaborated. It differs from the traditional ones by the complete exclusion of the salts-smelters. This method is based on the use of the influence of freezing and fine-dispersed comminution on the raw material. It allows open biological potential of the rennet cheeses more fully and to extract the hidden (bound) protein forms from nanocomplexes of lipids and mineral substances. It allows destruct the proteins of rennet cheeses to the separate polymers and dipeptides and tripeptides. The used technological methods gave a possibility to exclude the salts-smelters at the rennet cheeses manufacturing. They favor the transformation of lipid-proteins paracaseinate calcium phosphate complexes to the separate amino acids and peptides and allow receive homogenous plastic mass.It was established, that at the complex action of freezing and fine-dispersed comminution on the rennet cheese the destruction of hardly soluble lipid-protein nanocomplexes and release of protein from the bound state into free one – nanoform (by 33,5…35 % more) takes place. The mechanisms of this process, connected with cryomechanodestruction of connections between lipids and proteins and non-fermented catalysis of nanocomplexes were described.It was established, that cryomechanodestruction and non-fermented catalysis of protein to the separate monomers – α-amino acids (by 55…60 %) takes place at freezing and fine-dispersed comminution of rennet cheese before melting. The mechanism of freezing and non-fermented analysis, connected with cryomechanocracking of protein molecules at the expanse of peptide protein connections destruction to the separate α-amino acids and their transformation into the free form was described. It was also demonstrated, that the conformational changes of protein molecules take place synchronously.The offered and elaborated nanotechnology of melt cheese products on the base of rennet cheeses without salts-smelters includes complex action of freezing and fine-dispersed comminution. The mechanisms of processes, connected with cryomechanodestruction of connections between lipids and protein to the separate α-amino acids are described.The cheese fillings for “Pancake” confectionary and cheese snacks – falafels were manufactured on the base of cheese mass, received using the new method and enriching vegetable nanoadditives. They exceed the well-known analogs by chemical composition and are remarkable for the storage life, increased in 2 times. At the same time the significant part of substances (BAS and biopolymers) in cheese filings is in nanodimensional form (55…60 % of protein), especially, free α-amino acids, easily assimilated by the human organism. The recipes and technologies of sauces-dressings, sauces-deeps, cheese snacks and so on are also elaborated on the base of cheese mass, received by the new method

Technology of Healthy Processed Cheese Products Without Melting Salts with the Use of Freezing and Non-fermentative Catalysis

Authors studied comprehensive influence of the processes of non–fermentative catalysis – cryomechanolysis and freezing of solid rennet cheeses during their preparation for melting, which leads to the cryodestruction of low–soluble paracaseinatcalciumphosphate nanocomplexes into soluble gel form. It was established that there occurs their cryodestruction and transformation of their significant part to the nanoform (by 45…55 %). A nanotechnology of healthy processed cheese products was developed. Mechanisms of the processes were revealed. It was established that during freezing and finely dispersed grinding of solid rennet cheeses before melting, there occurs cryomechanodestruction and non–fermentative cryocatalysis (destruction) of protein molecules to separate monomers – α–amino acids by 55… 60 %, that is a significant part of amino acid is transformed from the bound state to the free soluble form. A mechanism of the process was revealed; it was shown that in parallel with the destruction of nanocomplexes, nanoassociants of protein, its conformational changes take place: erasing molecules, decreasing in volume, shape, the ratio of hydrophobic and hydrophilic groups in a molecule, and filling the nucleus of a molecule with hydrophobic residues.Authors proposed and developed the cryogenic nanotechnology of manufacturing processed cheese products based on solid rennet cheeses without melting salts, which includes an integrated influence of freezing and finely dispersed grinding, non–fermentative catalysis. It was established that cheese products, produced by the nanotechnology (fillings for confectionery products "PanCake", dressing sauces, dipping sauces, ball shaped snacks) and enriched with herbal additives, exceed the known analogs in chemical composition. In addition, a large part of substances (as BAS and biopolymers) in cheese products is in the nanostructured form (55...60 % of protein) in the form of free amino acids.New technologies of healthy processed cheese products have been tested under production conditions at a number of the Ukrainian enterprises (TOV VKG "Lisova kazka", NVP "FIPAR", NVP "KRIAS–1"). The regulatory documentation (TU, TI for "cheese and vegetable fillings for confectionery products "PanCake" and "cheese dressing sauces") was developed and approved

Tunability of DNA Polymerase Stability during Eukaryotic DNA Replication

2019 Elsevier Inc. Structural and biochemical studies have revealed the basic principles of how the replisome duplicates genomic DNA, but little is known about its dynamics during DNA replication. We reconstitute the 34 proteins needed to form the S. cerevisiae replisome and show how changing local concentrations of the key DNA polymerases tunes the ability of the complex to efficiently recycle these proteins or to dynamically exchange them. Particularly, we demonstrate redundancy of the Pol α-primase DNA polymerase activity in replication and show that Pol α-primase and the lagging-strand Pol δ can be re-used within the replisome to support the synthesis of large numbers of Okazaki fragments. This unexpected malleability of the replisome might allow it to deal with barriers and resource challenges during replication of large genomes

Mechanism of polymerase collision release from sliding clamps on the lagging strand

Replicative polymerases are tethered to DNA by sliding clamps for processive DNA synthesis. Despite attachment to a sliding clamp, the polymerase on the lagging strand must cycle on and off DNA for each Okazaki fragment. In the ‘collision release' model, the lagging strand polymerase collides with the 5′ terminus of an earlier completed fragment, which triggers it to release from DNA and from the clamp. This report examines the mechanism of collision release by the Escherichia coli Pol III polymerase. We find that collision with a 5′ terminus does not trigger polymerase release. Instead, the loss of ssDNA on filling in a fragment triggers polymerase to release from the clamp and DNA. Two ssDNA-binding elements are involved, the τ subunit of the clamp loader complex and an OB domain within the DNA polymerase itself. The τ subunit acts as a switch to enhance polymerase binding at a primed site but not at a nick. The OB domain acts as a sensor that regulates the affinity of Pol III to the clamp in the presence of ssDNA