Pectoskin

Pectoskin

The feasibility and scientific plausibility of the project are presumed to be discretionary. As a rule LP refrains from giving overall contextual connections in public digressions - in the interest of protecting expertise! Furthermore, this discretion should not, may not and cannot not be used for research purposes.

Zuckerrübe.jpg

Table of contents:  Executive Summary

I  Market evaluations

 

Preamble

Art. 1                       Competitive market 

Art. 2                       Highlights

Art. 3                       Opportunities and risks

Art. 4                       Raw material factor & equipment

II  Pectinization verification

 

Preamble

Art. 1                       Endo-polygalacturonase activity

Art. 2                       Short pectin anatomy

Art. 3                       Pectin production process

Art. 4                       Chemical structure of pectin

Art. 5                       Rhamnose sub-bond

Art. 6                       Gel formation according to degree of esterification

Art. 7                       Pectinolytic enzymes

Art. 8                       Enzyme nomenclature  

Art. 9                       Coenzyme

Art. 10                     Pectinesterases

Art. 11                     Extraction of pectinolytic enzymes

Art. 12                     Quality measures

Closing note          Pitch deck

tranparent.png

THE IP LEGAL DISCLAIMER: In order to preserve the "Intellectual Property" (IP), the overall presentation of the research details for the process engineering extraction and gelation of pectins from sugar beet pulp is not included. Only fragments that are structurally the same are presented, on the basis of which the research-scientific procedural steps can be reconstructed in a plausible way. Nevertheless, this sketch is subject to copyright protection. IP infringements are sanctioned in accordance with § 106 Copyright Act (UrhG) Section 78 para. 3 no. 4 German Criminal Code (StGB). © LP

Executive Summary

LP provides confectionery manufacturers with a new source of raw materials: Sugar beet pectin obtained from beet pulp. The term "pulp" refers to all water-insoluble constituents from beet that accumulate after extraction of the sugar-containing cell sap as cellulose, hemicellulose, (proto)pectin, etc. The latter transforms LP to gellable pectin– for the first time ever! As a note: 

 

Above all, gelatin production requires tryptophane-free proteins in the form of hydrolyzed collagen from mostly animal waste. However, the fact that the peptide chains used for this purpose are constructed in a left-turning helix molecular structure analogous to right-turning polysaccharide pectin chains prompted competitors to replace gelatin with pectins (with the exception of sugar beet - due to a lack of technical expertise). LP is changing this: using new microbial enzymatic techniques, LP ferments the protopectins, which have not yet been considered gellable anywhere, with the help of specially cultivated bacteria (in addition to specified yeasts), in such a way that they achieve the required elasticity and firmness of gummy bears. In order to achieve this to the necessary standards, knowledge in biology and microbiology as well as in molecular medicine and biochemistry is required, along with higher mathematics and corresponding physics. However, since universities and institutes generally do not conduct interdisciplinary research, such a comprehensive range of topics can only be addressed by a researcher covering these requirements. 

In the meantime, feasibility can be proven both scientifically and on the market. All necessary testing measures were pre-invested and implemented by LP:

 

  1. LP developed new microbial enzymatic processes for gelling sugar beet pectin.

  2. LP cultivated esterification-supporting microbes and bacteria for this purpose. 

  3. LP bred specified yeasts such as Saccharomyces parallel to step 2. 

  4. LP fermented these microbially-enzymatically prepared protopectins into gellable pectins.

  5. LP added starch-forming enzymes (potatoes, corn, etc.) to the "pectinization process"...

I  Market evaluations

 

Preamble

Over the past 200 years, so many durability methods have been developed that consideration for our health has remained neglected for far too long. This truth has caught up with us. This problem of humanity cannot be reduced in terms of demographic global requirements, nor should it be ignored: since the Neolithic Revolution over 7,000 years ago, humans have domesticated about 7,000 plants and almost the same number of animal species. For over 100 years, however, fewer and fewer different raw materials are used (currently only 12 crop species - one of which is sugar beet - and 14 livestock species cover over 98% of our global food requirements). Since this trend will increase, more attention should be paid to the remaining plants used for food, which LP is doing. 

 

Art. 1

Competitive market

No one in the world before LP has succeeded in making sugar beet pectin haptically gellable. The most common non-animal alternative to this kind of gelatin is algae. Apples, citrus fruits, and corn can nevertheless find a serious raw material niche here. Algae, however, are basically nothing more than fecal (colibacterial) binding products of mostly animal/human excrement. Furthermore, algae are generally decomposed by toxics in such a way that their processing into alternative gelatin is unhygienic. No industrial processor is sufficiently capable of cleaning this "fecal product" of toxic substances so precisely that it can be used with a clear conscience as an alternative raw material. Moreover, the fact that transformed sugar beet pectinization is not yet accessible to anyone else gives this world innovation a self-explanatory market advantage. In addition, the EU Sugar Regulation, which expired on September 30 2017, sent sugar beet market prices on a life-threatening downward spiral, making millions of tons of beet harvested each year worthless and superfluous – this trend is further increasing. The previously mindless devaluation of sugar beet will need to be reevaluated. 

 

Furthermore, the specific properties of this pectin, which has been decoded by LP for the first time, hold the potential of pH-balancing, immunomodulating, detoxifying, anti-inflammatory, antioxidative, and regenerative efficacy that is still untapped in the global cosmetics market - while the dermatological application of the beet pectin products prepared by LP does not require the use of synthetic substances. 

Art. 2

Highlights:

Thanks to microbial-enzymatic gelation methodology, the following additional specific property in the sugar beet pectins is derived: by means of bacteriological application steps (using lactic acid proteins adjusted to pH < 3.7), a large number of free valences are produced at the C41H60O36 chains, which are thus enabled to excessively uptake nutritionally significant vitamins. This is especially true with regard to the equally important health aspect! The lactic acid required for microbial-enzymatic gelling does not come from animal milk, but instead for example from glucose syrup from GMO-free corn. LP thus also fulfills the vegan criterion and at the same time excludes the allergen labeling that is subject to EU regulations. LP also optimally requires the integration of vitamins: on the C41H60O36 chains, lactic acid (C3H6O3) is used to produce a large number of free valences (Ø1:12!), resulting in above-average vitamin coupling. This non-animal lactic acid or, analogous to the above-mentioned C41H60O36 chains, right-turning lactic acid also achieves far higher congruence as a result... 

Art. 3

Risks and Opportunities

The market opportunities for the new raw material source to be developed, sugar beet (proto)pectin, are beyond question for the aforementioned reasons. In this respect, this market lead is only subject to risks if this "secret formula" is mishandled. This makes it all the more important to take into account the cooperation with companies that may be involved in research: LP will only permit the exchange of research/knowledge with partners who may be involved in the research if they do not have knowledge of the overall breakdown of the procedure. In practice, LP keeps the following confidential: all the microbes or bacteria, enzymes, yeasts etc. that LP uses in the microbial-enzymatic research and development process are the real secret to the mixture. As the "key/encryption supplier", LP personally and exclusively monitors compliance with industrial property rights. It should also be considered that the "swine flu" is breaking out with increasing frequency in different virus variations (A/H1N1, A/H1N2, A/H3N2, etc.) and "foot-and-mouth disease virus" (FMDV in a similarly different variation) is spreading visibly in cattle, along with what is generally known as the "avian influenza” (A/H5N1, A/H7N9, A/H5N8, etc.) – these are all animals from which gelatin is obtained for confectionery manufacturers, making the development of a new source of raw materials indispensable. It goes without saying that the aforementioned "fecal binding product" agar-agar is not used as an alternative raw material, in accordance with nutritional-physiological quality responsibility. Besides the comparatively small pectins from the peel of apples, citrus fruits, etc., sugar beet pectin is the only major alternative!

 

Art. 4

Raw material factor & equipment

According to fragmentary projections, beet pectin extraction and processing results in a raw material cost savings potential of >70% below that of conventional processes. In practice, the sugar beet/pulp processing required for this is carried out in the following order (simplified summary):

  

  1. Wheel loader fills truck with sugar beet/pulp.

  2. Truck unloads sugar beet/pulp into hopper via conveyor belt.

  3. Conveyor belt guides these via vibrating screen, on which water is pelting via pressure nozzles.

  4. Beets/pulp pre-cleaned in this way are transported from the vibrating screen to the coarse chaff mill. 

  5. Coarsely chopped material is ground by adding saponin-shifted NaCl solution by means of another knife mill.

  6. The result is then homogenized and ground again with a high-frequency mill. 

  7. Suspension is then mechanically stirred in large boilers for 2 hours.

  8. The contents of the boiler are pressed through nylon filter bags with a suction outlet.

  9. The remaining filter cake is again "wrung" from the filter bag. 

  10. Centrifuge the filtrate at 4°C.

  11. Clear supernatant is concentrated with ultrafiltration. 

  12. Enzyme solution, salts, reducing sugars and all what could still pass through the membrane, together with H2O, are removed. 

  13. As soon as the enzyme solution is concentrated, the membrane is rinsed with a NaCl solution (4°C) added to the retentate. 

  14. The remaining enzyme extract is freed from the soluble substances with low molecular mass by means of a dialysis bath.

  15. Dialysis solution = deionized water, NaCl and a very small amount of sodium azide (against microbial growth). 

  16. The enzyme extract is then passed through a semi-permeable membrane (a "dialysis tube" made of cellulose). 

  17. Dialysis tube is boiled before filling with enzyme extract for disinfection. 

  18. The filled dialysis tube is leaded (at constant 4°C) twice in dialysis solution for at least 4 hours each time. 

  19. The extract is centrifuged again to remove residual small precipitates.  

  20. The enzyme extract is then dried to powder.

  21. To begin with, the powder is cleanly temporarily stored.

  22. The pectins are then precipitated from this powder and prepared for microbial-enzymatic gelling...

  23. Pectins from the sugar beet pulp that can be gelled can then be used by confectionery manufacturers.

Steps 1-22 take place in a pectin factory. Step 23 concerns confectionery manufacturers. LP's part included: 

 

  1. LP developed microbial-enzymatic manufacturing processes for the industrial gelling of sugar beet pectins.  

  2. LP cultivated (using chemotaxonomic and phylogenetic identification steps) a large number of esterification-supporting bacteria, which together achieve an oxidizing effect in order to provide binding aids by means of free valences. 

  3. LP bred - parallel to TZ 2) - yeasts specified by means of microbiochemical convergence exclusion steps, e.g. Saccharomyces. 

  4. LP fermented the microbially-enzymatically prepared beet protopectins in TZ 2+3) (with the aid of the above-mentioned bacteria, yeasts etc.) to gellable pectins. However, this was no longer a matter of fundamental proof of feasibility, but instead of process detailing of a) efficiency of raw material yield, b) determination of the equipment required for industrial production and c) overall cost optimization. With step in TC 4), the target result of the steps in TC 2+3) was already reached as the basis for the subsequent step in TC 5). 

  5. LP added starch-forming enzymes (from potatoes, corn, etc.) to the pectinization process in order to achieve the required elasticity and stability with their supplementary aid. 

II. Pectinization verification

Preamble:

People prefer more energy-rich foods to more natural ones. In evolutionary history, this is mainly due to the great trauma called and , which has been going on throughout human history. In addition, the oversized human brain devours an enormous amount of energy. The energy used in the search for food has always had to produce correspondingly higher yields as a result. Conclusion: nutritionally, humans prefer to consume more energy-rich / animal proteins vs. digestion-friendly / plant proteins. This also applies to animal gelatin from various confectionery manufacturers. In times of climate change and global environmental pollution, it is now only logical to promote vegetarian food, while reducing our meat consumption to a healthier level. This is against the backdrop of a shortage in animal raw materials and an increase in raw material prices. Why and how LP uses sugar beet as a new source of alternative raw materials for confectionery manufacturers is highlighted in the following side note. 

 

Art. 1

Ando-polygalacturonases activity

The “pectin” ingredient in sugar beet is relevant here. It is obtained, for example, from press shreds from beet sugar production. After this has been turned into thick juice from biogas, the thick juice contains plenty of pectins, so an economic interest in its gellable usability is self-evident. The quantification of pectinolytic enzymes in sugar beets is essential to determine how active they are. Since the activity of the endo-polygalacturonases breaks down the long-chain pectins into short-chain pectin fragments, it is precisely this property that makes sugar beet pectins unsuitable, since they exhibit such altered gelling properties that their viscosity (even in aqueous solutions) does not permit gelling in this way. An important Casus Cnactussubdues the endo-polygalacturonase activity (e.g. when storing thick juice). The comparatively low degree of esterification of sugar beet pectin is therefore partly due to pectinesterase activity. Conversely, this means: just as it is possible to successively reduce the desesterification step by adding microbial pectinesterase removal enzymes when the plant's own pectinesterase has a correspondingly high activity, it should also be possible to increase the degree of esterification by adding microbial pectinesterase removal enzymes (with the aid of bacteria cultivated specifically for this purpose and special yeasts), insofar as this reduces the activity of the endo-polygalacturonase. The pH value is particularly important here, since, for example, when the thick juice is stored, lactic acid fermentation occurs, which causes the pH value to drop to 3½. Our research therefore particularly focuses on the sugar beet enzymes, as they are essentially responsible for the degradation of sugar beet pectins. This means it is necessary to find out how active these enzymes areand under which conditions they optimally degrade pectin, in order to find out how this process can be counteracted enzymatically and microbially.

Art. 2

Short pectin anatomy

Besides gelatin, pectin is the high-molecular natural product par excellence. It has gel-forming properties and is primarily used as a hydrocolloid. Pectins themselves are highly branched polysaccharides which occur in all higher land plants as so-called "structural elements". Pectins are located in the middle lamella, a layer between the cells and cell clusters, where they hold the cells together as a kind of "cement substance". In the aforementioned middle lamella they form, together with hemicellulose and glycoprotein, an amorphous matrix. Cellulose microfibrils are then embedded in it. The decisive factor here is that Ca++ bridges are formed between the individual pectin molecules, without which no interconnection of the pectin molecules would otherwise be possible (see fig.).

pectin.png

Art. 3

Pectin production process

To date, there are two larger process groups for pectin production. The larger one is conventional: the acid hydrolysis by means of sulphuric acid solution causes the otherwise insoluble protopectin to be dissolved out of the cell walls of the starting materials. The extracted pectin solution is then filtered and evaporated. It is then precipitated by adding alcohol (e.g. isopropanol). The raw pectin is produced, which is then dried, ground and homogenized. Furthermore, the extracted pectins are precipitated with aluminum salts, or the dried plant-based residue is treated with gaseous ammonia in order to extract the "amidated" pectins with water (or a strongly diluted lye). These are finally precipitated by means of an acid. The second major process group is the biotechnological process in which enzymes are used to extract and modify the otherwise insoluble protopectin from the middle lamella. The enzymes used here are called "protopectinases" and are obtained from microorganisms (Candida guilliermondi, Bacillus licheniformis spore amongst others).Compared to the conventional traversing group mentioned above, a significantly higher yield and lower energy consumption are achieved. Also, in doing so, no nitrate-loaded pomace is produced. This so-called biotechnological production process, however, influences the chemical structure of the pectin and modifies its properties as a result; nevertheless, this process is of particular research relevance for LP. LP extracts the pectin purelyphysically... 

 

[The know-how of the procedure developed by LP is only shown in a plausible way and is completely subject to secrecy.]

 

Art. 4

The chemical structure of pectin

The chemical structure of pectin is of particular importance: similar to the collagen structure, pectin is determined by the plant species, as well as (and this is particularly noteworthy) the respective plant component, the plant age itself, plant cultivation and breeding. Pectin generally belongs to the large group of polysaccharides. According to its basic structure, it usually consists of α-1-4-glycosidically linked D-galacturonic acid molecules, which are esterified with methanol amongst others. Plant regions, in which 

almost only galacturonic acid molecules are anchored, (see schem. Fig. r.) are regarded as unbranched and are called homogalacturans. Only 1 rhamnose unit (C6H12O5) comes out of about 200 galacturonic acid units. On the other hand, the strongly branched regions (“hairy regions”) are built up in the backbone of the pectin molecule, alternating between α-1-4-linked D-galacturonic acid molecules and α-1-2-linked L-rhamnoses. The side chains of the "hairy

pectin 2.jpg
pectin 3.png

regions" are formed from glycosidically bound D-galactoses, L-arabinoses and D-xyloses, which give them high resistance to pectinolytic enzymes. Hairy regionsaccount for max. 5% of the total galacturic acid content of the pectin molecule. In the latter, the galacturonic acid units are rotated by 120° against each other, so that every fourth takes the same position. On the other hand, the dissolved molecule takes on a helical structure in which 3 galacturonic acid units form 1 complete turn (by virtue of steric factors and above all, stabilizing hydrogen bridge bonds). 

Art. 5

Rhamnose Sub-bond

As a rule, two neighboring helixes (in the solution) are arranged exactly parallel to one another. But the regular helix structure is interrupted by the rhamnoses, which then leads the linear pectin structure to buckle and thus negatively influences the hydration ability and the gelling properties. The macromolecular dimerization of the pectin chains influenced in this way disturbs the formation of important adhesion zones and favors the immobilization of solvents (usually water). As a result, the rhamnoses responsible for this must be made enzymatically ineffective. This is because in the end: For gel-forming properties, the degree of galacturonic acid molecule esterification of the carboxyl groups is decisive. However, these are partly neutralized with methanol and partly (or completely) neutralized by negative cations (e.g. Na+and Ca++). Pectins are high and/or low methylated. (Highly methylated pectins have a degree of esterification of more than 50%). There are no further subdivisions; only the group of medium methylated pectins, with a degree of esterification that varies between 45 - 65%, should be mentioned briefly. A detailed description of the enzymatic-microbial rhamnose sub-bond is left out due to IP.

Art. 6

Gel formation according to the degree of esterification

During gel formation, the degree of esterification of the pectin is closely related to the gel formation rate and the texture of the gels themselves. In other words: highly methylated pectins gel faster, and do this at higher temperatures. To form the gel itself, a sugar content of at least 55% by weight and a pH value of 1 to 3½ is required. The acid ensures that the pectins are not negatively charged and repel each other as a result. In the meantime, the sugar binds the water, which would otherwise be absorbed by pectins. In this way, binding between the molecules is prevented. If the ratio of pH-value and acidity is correctly adjusted, a sufficient number of pectin molecules appear, after which the pectin chains can be connected with one another (with the help of corresponding hydrogen bonds). Pectins with a degree of esterification below 50% require the additional use of calcium, but sugar is no longer needed. In this reaction, the pH range (by adding calcium) is between 1 and 7. Pectic acid itself is pectin with a degree of esterification of less than 5%. However, pectic acid is able to form gels under the same conditions (equal to the low methylester pectins). At high pH values (due to the high content of polyvalent cations) the pectic acid even precipitates as pectate (salts of unesterified pectin acids). However, contrary to general assumptions, sugar beet pectic acid (C41H60O36) has theproperty of being prepared enzymatically and microbially in such a way that it is able to form solid gels (like higher methylester pectins). Details are also left out here due to IP.  

 

Art. 7

Pectinolytic enzymes

Enzymes are bio/protein polymers, which are formed by cells and work as so-called "biocatalysts". They consist of one or more subunits composed of amino acids (20 in total). These in turn form a protein-specific sequence and are linked to each other via peptide bonds. Peptide bonds in turn are formed by carboxyl/amino groups of individual amino acids (with elimination of water). If there is a compound of more than 100 amino acids, it is referred to as a protein. If the compound contains between 10 and 100 amino acids, it is considered a polypeptide. In the enzyme, proteins take on a specific structure and are spatially arranged so as to form a three-dimensional structure with a pocket-like depression (the center of action). This specific, 3-dimensional structure is a prerequisite for enzymes being able to form an "enzyme-substrate complex" with very specific substances (referred to as substrates). The dynamic, spatial arrangement in the action center determines the "key-lock principle" of the enzyme. The power of this arrangement hardly requires any activation energy, which leads to an immense increase in the speed of the actual reaction process. Bio/chemical reactions are therefore increased 1010 - to 1020 - fold. The molecular weight of each enzyme is between 10,000 and several million daltons. However, these enzymatically catalyzed reactions are also subject to typical saturation kinetics. Since enzymes (in comparison to chemical catalysts) can only catalyze one or very few reactions at a time, one speaks of reaction-specific catalysts, whereby the specificity of this enzyme catalysis does not concern the substrate as a whole molecule, but only the chemical groups of the substrate. This makes the enzymatic research of LP all the more precise.

 

Art. 8

Enzyme nomenclature 

With regard to their nomenclature, it is also noticeable that for a large number of different enzymes several names are often used at the same time (which is certainly due to the fact that earlier enzymology arbitrarily gave the enzymes names). It was not until much later that enzyme names were created by adding the suffix "-ase" to the substrate converted by the respective enzyme. As a result, starch-splitting enzymes were soon called "amylases" and fat-splitting "lipases". According to their respective functionality, various enzyme groups also received trivial names such as "oxidases", "dehydrogenases", "decarboxylases", some of which have survived to this day. A precise nomenclature is provided by the IUB, which describes the mechanism of the reaction more precisely... Further details are omitted, since the addressee(s) may also be familiar with the complex enzyme classification. According to IUB, each enzyme is assigned a key number, which is identified as an EC number, and its first number reveals the main class of enzymes. There are a total of six main enzyme groups. For example, the pectinesterase has the key number "PE, EC 3.1.1.11", where the 3 stands for the third main group and therefore indicates that this enzyme is a hydrolase. Cofactors or coenzymes in enzymatic reactions are no less important and the non-protein molecules are mainly required by the enzymes of main groups 1, 2, 5 and 6. 

Art. 9

Coenzymes

Coenzymes that participate in reactions (like a 2nd substrate) are co-substrates. One of the most important coenzymes is the NADP+molecule, since it is hydrogen-transferring. These coenzymes are usually firmly linked to the actual enzyme, and are occasionally also linked by covalent bonds with the protein. The resulting enzyme and coenzyme complex is called a holoenzyme, which has a protein component called an apoenzyme. This presence of coenzymes is therefore decisive for enzyme activity (cf. Art. 1, endo-polygalacturonase activity control). There are still many other factors influencing enzyme activity (pH value, concentration of dissolved salts, reaction temperature, pressure, etc.), which are assumed to be known to the addressee(s). As a result, no further details are provided. It should also be noted that by quantifying the increase in reaction product of the decisive pectinolytic enzyme "exo-polygalacturonase” also determines the activity of this enzyme by photometrically measuring it, in addition to pectin esterase and endo-14-polygalacturonase, which are determined on the basis of its activity.

Art. 10

Pectin esterases

Pectin esterases, also known as pectin methyl esterases, like polygalacturonases belong to the large main group of hydrolases. On closer inspection, however, they belong to the subgroup of carboxyester hydrolases. Of course, these enzymes hydrolyze the methyl ester groups of pectin. As a result, high methylester pectin results in low methylester pectin, which is formed by the action of pectin methyl esterases on the methyl ester groups of the pectin to form pectic acid and methanol. In this case as well, LP (in the logical reverse) enzymatic-microbially transfers the low esterification into high esterification. As usual, IP-related details are also (and especially here) omitted. Only this in advance: the polygalacturonases catalyze the cleavage of the α-1-4 glycosidic bonds between the galacturonic acid molecules of the pectin. The endo-polygalacturonase (EC 3.2.1.15) cleaves the pectin chain quite randomly, whereas exo-polygalacturonases (EC 3.2.1.67) cleaves monogalacturonic acid units (dimers) from the non-reducing end of the chain. (Exo-polygalacturonases and endo-polygalacturonases occur in all higher plants and numerous microorganisms). Furthermore, since the formation of an enzyme-substrate complex can only occur in the immediate proximity, free cayrboxyl groups are used without exception via hydrolysis. Along the pectin chain, therefore, ever larger sections are formed in which pectin is de-esterified - a basic prerequisite for further hydrolytic cleavage by polygalacturonases! In accordance with the enzymatic "key-lock-principle", the reverse conclusion is also drawn here from de-esterification to esterification (which in practice is much more complicated than it sounds here). 

 

Art. 11

Extraction of pectinolytic enzymes

Pectinolytic enzymes occur only very scarcely in plant tissue. This makes their correct extraction process all the more important, which usually takes place in the following two ways: the first is by precipitating the enzymes using ammonium sulphate, whereas LP prefers ultrafiltration, since this is not so time-consuming and also has a much lesser effect on enzyme activity. However, both manufacturing processes must be carried out in all their steps at 4°C, because enzymes are inherently thermally unstable. Enzyme extraction through ultrafiltration takes place in three process steps: 

  1. Digestion of the cell tissue (plant tissue is transformed with the addition of saponin-transferred NaCl solution using a knife mill). Homogenization and further, finer transformation of the plant tissue pieces, e.g. using Ultra-Turrax. The suspension is stirred for two hours, and then passed through a nylon cloth or similar to separate the remaining components for the first time; the remaining filter cake is then wrung out with a nylon cloth. Centrifuge filtrate at 4°C. 

  2. Clear supernatant is concentrated with ultrafiltration; enzyme solution then becomes salts, reducing sugars and any substances that could still pass through the membrane, together with water. As soon as the enzyme solution is concentrated to a certain volume, the membrane is rinsed, to the intended degree, with a NaCl solution added to the retentate and tempered to 4°C. 

  3. The remaining enzyme extract is freed from low molecular weight soluble substances by means of a dialysis bath containing a dialysis solution of deionized water, NaCl and very low sodium azide. Sodium azide is meant to prevent microbial growth. The enzyme extract is located underneath in a semi-permeable membrane (cellulose "dialysis tube"). The dialysis tube is boiled before filling with enzyme extract for disinfection. Filled dialysis tube leaded (at constant 4°C) twice in dialysis solution for at least 4 hours. Extract is centrifuged again to remove residual small precipitates. 

 

Finally, the enzyme extract can be used to determine the activity of the endo-polygalacturonases and pectinesterases (cf. again Art.1, casus cnactus)! Furthermore, the consideration of the colloidal influencing parameters is considered an essential measurement criterion. 

Art. 12

Quality measurabilities

State-of-the-art rapid methods for the analysis and guarantee of food quality increasingly require the search for chemical-physical quantities that can be measured as quickly as possible and reliably meet the quality requirements. One of the most recent methods is rheological (rheology = science of material deformation/flow behavior, sub-discipline of elasticity/plasticity theory together with fluid mechanics of non-Newtonian fluids). In general, it is responsible for the research of "continuum mechanical" problems, i.e. the evaluation and conclusion of material laws required for this purpose, namely from the microscopic or nanological basic structure of different material precipitates (suspensions, macromolecular systems, etc.). 

 

However, the general literature so far hardly reveals any material-scientific information on the fluid-dynamic behavior of raw materials and intermediate/final/byproducts of all industrial manufacturing processes. As a result, LP attached particular importance to an all-encompassing, material-scientific examination of the processing technologies. Before LP there was already marginalresearch work on the fluid dynamic behavior of concentrated solutions in the evaporation station, although the fluid dynamic characteristic values of the intermediate products were largely only determined by LP: Its material scientific and rheological investigations led to some unexpected conclusions about the chemical and physical processes in, for example, subsequent process steps:

 

  1. Determination of physical-mechanical properties of the raw material in relation to ingredients.

  2. Evaluation of the rheological behavior of native extraction juices directly after crushing beets.

  3. Creation of a viscosity matrix for the processing of aged beets.

  4. Determination of the rheological combination behavior of pure molasses, vinasse, sucrose solution... as a
    function of temperature (30 to 130°C), dry matter and lime salt content, pH value, etc.

  5. Rheological characterization of molasses and vinasse in relation to dry matter and their chemical consistency. 

  6. Determination of the influence of viscosity by pectin and dextran in raw extract. 

 

Furthermore, since the rheological behavior of vinasse (a derivative of molasses fermentation) prior to LP has hardly been investigated, LP was the first to find out that the dependence of the flow behavior (Newton/non-Newton) is largely determined by the previous process – but not by the dry matter. LP also found out that – contrary to expectations – density and specific heat capacity of the thermal conductivity coefficient (together with specific boiling point increase) do not necessarily have to be taken as a basis for the thermal physical characteristic values, which considerably simplifies the process overall. However, the thermal-physical characteristic values served LP as orientation data for the rheological vinasse measurement that it had newly developed.

pectin 4.png

Closing note: 

The confectionery industry is in a raw material dilemma due to global consumer demand. This applies in particular to confectionery manufacturers of raw materials that feed on animal waste products for gelatin. For example, the miserable attempt to compensate for the shortage of animal raw materials with alternative plant sources (pectins from the peels of citrus fruit, apples, etc.) is evidence of this. On the other hand, the blatant use of algal pectins (agar-agar) on the market is nutritionally pure imposition and thus irresponsible. But sugar beet pectins are both healthy and health-promoting. LP holds the know-how monopoly!

©LP, Oct 20, 2017