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Xylanase Enzyme
Pulp Bleaching / Silage / Anaerobic digestion / Poultry farming / Wheat flour / Dough conditioner / Decaffeination / Clarifying agent / Degumming / Pectinase / Pectin / Cellulase

Knowledge base :

Xylanase, Xylan, Antinutrient, Hemicellulose, Xylose, Furfural, Bleaching of wood pulp, Silage, Anaerobic digestion, Poultry farming, Wheat flour, Dough conditioner, Decaffeination, Clarifying agent, Introduction to Degumming, The Nature of Gums and Phosphatides, Pectinase, Pectin, Cellulase


Overall structure of a family 10 xylanase (2F8Q) showing the typical TIM-barrel fold (a) top (b) side view.

Xylanase (EC 3.2.1.8)  is any of a class of enzymes that degrade the linear polysaccharide xylan into xylose,[1] thus breaking down hemicellulose, one of the major components of plant cell walls.

As such, it plays a major role in micro-organisms thriving on plant sources for the degradation of plant matter into usable nutrients. Xylanases are produced by fungi, bacteria, yeast, marine algae, protozoans, snails, crustaceans, insect, seeds, etc.;[2] mammals do not produce xylanases. However, the principal commercial source of xylanases is filamentous fungi.[2]

Commercial applications for xylanase include the chlorine-free bleaching of wood pulp prior to the papermaking process, and the increased digestibility of silage (in this aspect, it is also used for fermentative composting).[3]

Apart from its use in the pulp and paper industry, xylanases are also used as food additives to poultry; in wheat flour for improving dough handling and quality of baked products [1]; for the extraction of coffee, plant oils, and starch; in the improvement of nutritional properties of agricultural silage and grain feed; and in combination with pectinase and cellulase for clarification of fruit juices and degumming of plant fiber sources such as flax, hemp, jute, and ramie. A good quantity of scientific literature is available on key features of xylanase enzymes in biotechnology ranging from their screening in microbial sources to production methods, characterization, purification and applications in commercial sector.


  • Structure of xylan in hardwood

Xylan (/ˈzlən/) (CAS number: 9014-63-5) is a group of hemicelluloses that represents the third most abundant biopolymer on Earth. It is found in plants, in the secondary cell walls of dicots and all cell walls of grasses.
Xylans are polysaccharides made up of β-1,4-linked xylose (a pentose sugar) residues with side branches of α-arabinofuranose and α-glucuronic acids and contribute to cross-linking of cellulose microfibrils and lignin through ferulic acid residues. On the basis of substituted groups xylan can be categorized into three classes i) glucuronoxylan (GX) ii) neutral arabinoxylan (AX) and iii) glucuronoarabinoxylan (GAX).

Xylan is used in different ways as part of our daily lives. For example, the quality of cereal flours and the hardness of dough are largely affected by the amount of xylan thus, playing a significant role in bread industry. The main constituent of xylan can be converted into xylitol (a xylose derivative) which is used as a natural food sweetener, which helps to reduce dental cavities and acts as a sugar substitute for diabetic patients. It has many more applications in the livestock industry, because poultry feed has a high percentage of xylan. Some macrophytic green algae contain xylan (specifically homoxylan) especially those within the Codium and Bryopsis genera where it replaces cellulose in the cell wall matrix. Similarly, it replaces the inner fibrillar cell-wall layer of cellulose in some red algae.

Xylan is one of the foremost anti-nutritional factors in common use feedstuff raw materials. Xylooligosaccharides produced from xylan are considered as "functional food" or dietary fibers due their potential prebiotic properties. Xylan can be converted in xylooligosaccharides by chemical hydrolysis using acids or by enzymatic hydrolysis using endo-xylanases. Some enzymes from yeast can exclusively converts xylan into only xylooligosaccharides-DP-3 to 7.

Xylan is a major components of plant secondary cell walls which is a major source of renewable energy especially for second generation biofuels. However, xylose (backbone of xylan) is a pentose sugar that is hard to ferment during biofuel conversion because microorganisms like yeast cannot ferment pentose naturally.

 

Antinutrient  are natural or synthetic compounds that interfere with the absorption of nutrients.Nutrition studies focus on these antinutrients commonly found in food sources and beverages.

Phytic acid has a strong binding affinity to minerals such as calcium, magnesium, iron, copper, and zinc. This results in precipitation, making the minerals unavailable for absorption in the intestines. Phytic acids are common in the hulls of nuts, seeds and grains and of great importance in agriculture animal nutrition and eutrophication-wise due to the mineral chelation and bound phosphates released into the environment. Without the need to use milling to reduce phytate (including nutrient), the amount of phytic acid is commonly reduced in animal feeds by adding histidine acid phosphate type of phytases to them.

Protease inhibitors are substances that inhibit the actions of trypsin, pepsin and other proteases in the gut, preventing the digestion and subsequent absorption of protein. For example, Bowman–Birk trypsin inhibitor is found in soybeans.

Lipase inhibitors interfere with enzymes, such as human pancreatic lipase, that catalyze the hydrolysis of some lipids, including fats. For example, the anti-obesity drug orlistat causes a percentage of fat to pass through the digestive tract undigested.

Amylase inhibitors prevent the action of enzymes that break the glycosidic bonds of starches and other complex carbohydrates, preventing the release of simple sugars and absorption by the body. Amylase inhibitors, like lipase inhibitors, have been used as a diet aid and obesity treatment. Amylase inhibitors are present in many types of beans; commercially available amylase inhibitors are extracted from white kidney beans.




Phytic acid (deprotonated phytate anion in the picture) is an antinutrient that interferes with the absorption of minerals from the diet.

Most common molecular motif of hemicellulose

Hemicellulosee, A hemicellulose (also known as polyose) is one of a number of heteropolymer (matrix polysaccharides), such as arabinoxylans, present along with cellulose in almost all terrestrial plant cell walls.[1] While cellulose is crystalline, strong, and resistant to hydrolysis, hemicelluloses have random, amorphous structure with little strength. They are easily hydrolyzed by dilute acidor base as well as a myriad of hemicellulase enzymes.

Diverse kinds of hemicelluloses are known. Important examples include xylanglucuronoxylanarabinoxylanglucomannan, and xyloglucan.

Hemicelluloses are polysaccharides often associated with cellulose, but cellulose and hemicellulose have distinct compositions and structures. Diverse sugars comprise hemicellulose, whereas cellulose is derived exclusively from glucose. For instance, besides glucose, sugar monomers in hemicelluloses can include the five-carbon sugars xylose and arabinose, the six-carbon sugars mannose and galactose, and the six-carbon deoxy sugar rhamnose. Hemicelluloses contain most of the D-pentose sugars, and occasionally small amounts of L-sugars as well. Xylose is in most cases the sugar monomer present in the largest amount, although in softwoods mannose can be the most abundant sugar. Not only regular sugars can be found in hemicellulose, but also their acidified form, for instance glucuronic acid and galacturonic acid can be present.

In the 
sulphite pulp process the hemicellulose is largely hydrolysed by the acid pulping liquor ending up in the brown liquor where the fermentable hexose sugars (around 2%) can be used for producing ethanol. This process was primarily applied to calcium sulfite brown liquors.


d-Xylose

Xylose, (cf. Greek: ξύλον, xylon, "wood") is a sugar first isolated from wood, and named for it. Xylose is classified as a monosaccharide of the aldopentose type, which means that it contains five carbon atoms and includes an aldehyde functional group. It is derived from hemicellulose, one of the main constituents of biomass. Like most sugars, it can adopt several structures depending on conditions. With its free aldehyde group, it is a reducing sugar.

The acid-catalysed degradation of hemicellulose gives furfural,[6] a precursor to synthetic polymers and to tetrahydrofuran.
Xylose is metabolised by humans, although it is not a major human nutrient and is largely excreted by the kidneys.[8] Humans can obtain xylose only from their diet. An oxido-reductase pathway is present in eukaryotic microorganisms. Humans have enzymes called protein xylosyltransferases (XYLT1, XYLT2) which transfer xylose from UDP to a serine in the core protein of proteoglycans.
Xylose contains 2.4 calories per gram
[citation needed] (lower than glucose or sucrose, approx. 4 calories per gram).
Reduction of xylose by catalytic hydrogenation produces the sugar substitute xylitol.

Furfural is an organic compound with the formula C4H3OCHO. It is a colorless liquid, although commercial samples are often brown. It has an aldehyde group attached to the 2-position of furan. It is a product of the dehydration of sugars, as occur in a variety of agricultural byproducts, including corncobs, oat, wheat bran, and sawdust. The name furfural comes from the Latin word furfur, meaning bran, referring to its usual source. Furfural is only derived from lignocellulosic biomass, i.e. its origin is non-food or non-coal/oil based. Aside from ethanol, acetic acid and sugar it is one of the oldest renewable chemicals. It is also found in many processed foods and beverages.
Furfural may be obtained by the acid catalyzed dehydration of 5-carbon sugars (pentoses), particularly xylose.
C
5
H
10
O
5
  C
5
H
4
O
2
 + 3 H
2
O

These sugars may be obtained from hemicellulose present in lignocellulosic biomass, which can be extracted from most terrestrial plants.
It is found in many foods: coffee (55–255 mg/kg) and whole grain bread (26 mg/kg).

Furfural is an important renewable, non-petroleum based, chemical feedstock. It can be converted into a variety of solvents, polymers, fuels and other useful chemicals by a range of catalytic reductions.

Hydrogenation of furfural provides furfuryl alcohol (FA), which is used to produce Furan resins, which are exploited in thermoset polymer matrix composites, cements, adhesives, casting resins and coatings. Further hydrogenation of furfuryl alcohol leads to tetrahydrofurfuryl alcohol (THFA), which is used as a solvent in agricultural formulations and as an adjuvant to help herbicides penetrate the leaf structure.

Another important solvent made from furfural is methyltetrahydrofuran. Furfural is used to make other furan derivatives, such as furoic acid, via oxidation, and furan itself via palladium catalyzed vapor phase decarbonylation.

Furfural is also a specialized chemical solvent.
There is a good market for value added chemicals that can be obtained from furfural.



Furfural


Worldwide pulp production by type of bleaching used: Chlorine (Cl2), Elemental Chlorine Free (ECF) and Total Chlorine Free (TCF).. 

Bleaching of wood pulp is the chemical processing of wood pulp to lighten its color and whiten the pulp. The primary product of wood pulp is paper, for which whiteness (similar to, but distinct from brightness) is an important characteristic.These processes and chemistry are also applicable to the bleaching of non-wood pulps, such as those made from bamboo or kenaf.

Brightness is the amount of incident light reflected from paper under specified conditions,[2] usually reported as the percentage of light reflected, so a higher number means a brighter or whiter paper. In the US, the TAPPI T 452 or T 525 standards are used. The international community uses ISO standards. Table 1 shows how the two systems rate high brightness papers, but there is no simple way to convert between the two systems because the test methods are so different. The ISO rating is higher and can be over 100. This is because contemporary white paper incorporates fluorescent whitening agents (FWA). Because the ISO standard only measures a narrow range of blue light, it is not directly comparable to human vision of whiteness or brightness.


Silage is a type of fodder made from green foliage crops which have been preserved by acidification, achieved through fermentation. It can be fed to cattle, sheep and other such ruminants (cud-chewing animals). The fermentation and storage process is called ensilage, ensiling or silaging, and is usually made from grass crops, including maize, sorghum or other cereals, using the entire green plant (not just the grain). Silage can be made from many field crops, and special terms may be used depending on type: oatlage for oats, haylage for alfalfa (haylage may also refer to high dry matter silage made from hay).

Silage can be made by one or more of the following methods: placing cut green vegetation in a silo or pit; piling the vegetation in a large heap and compressing it down so as to purge as much oxygen as possible, then covering it with a plastic sheet; or by wrapping large round bales tightly in plastic film.

Ensilage can be substituted for root crops. Bulk silage is commonly fed to dairy cattle, while baled silage tends to be used for beef cattlesheep and horses. The advantages of silage as animal feed are several:

  • During fermentation, the silage bacteria act on the cellulose and carbohydrates in the forage to produce volatile fatty acids (VFAs), such as aceticpropioniclactic, and butyric acids. By lowering pH, these create a hostile environment for competing bacteria that might cause spoilage. The VFAs thus act as natural preservatives, in the same way that the lactic acid in yogurt and cheese increases the preservability of what began as milk, or vinegar (dilute acetic acid) preserves pickled vegetables. This preservative action is particularly important during winter in temperate regions, when green forage is unavailable.
  • When silage is prepared under optimal conditions, the modest acidity also has the effect of improving palatability and provides a dietary contrast for the animal. (However, excessive production of acetic and butyric acids can reduce palatability: the mix of bacteria is ideally chosen so as to maximize lactic acid production.)
  • Several of the fermenting organisms produce vitamins: for example, lactobacillus species produce folic acid and vitamin B12.
  • The fermentation process that produces VFA also yields energy that the bacteria use: some of the energy is released as heat. Silage is thus modestly lower in caloric content than the original forage, in the same way that yogurt has modestly fewer calories than milk. However, this loss of energy is offset by the preservation characteristics and improved digestibility of silage.


MB Trac rolling a silage heap or "clamp" in Victoria, Australia.


Anaerobic lagoon and generators at the Cal Poly Dairy, United States

Anaerobic digestion is a sequence of processes by which microorganisms break down biodegradable material in the absence of oxygen.[1] The process is used for industrial or domestic purposes to manage waste or to produce fuels. Much of the fermentation used industrially to produce food and drink products, as well as home fermentation, uses anaerobic digestion.

The most important initial issue when considering the application of anaerobic digestion systems is the feedstock to the process. Almost any organic material can be processed with anaerobic digestion; however, if biogas production is the aim, the level of putrescibility is the key factor in its successful application.[48] The more putrescible (digestible) the material, the higher the gas yields possible from the system.

Feedstocks can include biodegradable waste materials, such as waste paper, grass clippings, leftover food, sewage, and animal waste.

Poultry farming is the form of animal husbandry which raises domesticated birds such as chickens, ducks, turkeys and geese to produce meat or eggs for food. Poultry – mostly chickens – are farmed in great numbers. More than 60 billion chickens are killed for consumption annually. Chickens raised for eggs are known as layers, while chickens raised for meat are called broilers.

According to the World Watch Institute, 74 percent of the world's poultry meat, and 68 percent of eggs are produced intensively.
[6] One alternative to intensive poultry farming is free-range farming using lower stocking densities. Poultry producers routinely use nationally approved medications, such as antibiotics, in feed or drinking water, to treat disease or to prevent disease outbreaks. Some FDA-approved medications are also approved for improved feed utilization.


Bank of cages for layer hens


Wheat flour

Wheat flour is a powder made from the grinding of wheat used for human consumption. Wheat varieties are called "soft" or "weak" if gluten content is low, and are called "hard" or "strong" if they have high gluten content. Hard flour, or bread flour, is high in gluten, with 12% to 14% gluten content, and its dough has elastic toughness that holds its shape well once baked. Soft flour is comparatively low in gluten and thus results in a loaf with a finer, crumbly texture. Soft flour is usually divided into cake flour, which is the lowest in gluten, and pastry flour, which has slightly more gluten than cake flour.

In terms of the parts of the grain (the grass fruit) used in flour—the endosperm or protein/starchy part, the germ or protein/fat/vitamin-rich part, and the bran or fiber part—there are three general types of flour. White flour is made from the endosperm only. Brown flour includes some of the grain's germ and bran, while whole grain or wholemeal flour is made from the entire grain, including the bran, endosperm, and germ. Germ flour is made from the endosperm and germ, excluding the bran.

A Dough conditioner is any ingredient or chemical added to bread dough to strengthen its texture or otherwise improve it in some way. Dough conditioners may include enzymes, yeast nutrients, mineral salts, oxidants and reductants, and emulsifiers.


Examples of dough conditioners include ascorbic acid, distilled monoglycerides, citrate ester of monoglycerides, diglycerides, ammonium chloride, 
enzymes, diacetyl tartaric acid ester of monoglycerides or DATEM, potassium bromate, calcium salts such as calcium iodate, L-cystine, L-cysteine HCl, glycerol monostearate, azodicarbonamide,[5][6] sodium stearoyl lactylate, sucrose palmitate or sucrose ester, polyoxyethylene sorbitan monostearate or polysorbate, soybean lecithin, and soybean lecithin enriched with lysophospholipids.
Less processed dough conditioners include sprouted- or malted-grain flours, soy, milk, wheat germ, eggs, potatoes, gluten, yeast, and extra kneading. Malted, diastatic flours are not typically added by manufacturers to whole-wheat flours. Robertson et al. point out that some of the better information is found in baking books published back when bakers were still kneading by hand.



Freshly mixed dough in the bowl of a stand mixer


A cup of black coffee

Decaffeination .Decaffeination is the removal of caffeine from coffee beans, cocoa, tea leaves, and other caffeine-containing materials. Decaffeinated drinks contain typically 1–2% of the original caffeine content, and sometimes as much as 20%. Decaffeinated products are commonly termed decaf.
To ensure product quality, manufacturers are required to test the newly decaffeinated coffee beans to make sure that caffeine concentration is relatively low. A caffeine content reduction of at least 97% is required under United States standards. Less than 0.1% caffeine in decaffeinated coffee and less than 0.3% in decaffeinated instant coffee in Canada.


Decaffeination of tea and coffee is also an example of an extraction, where the caffeine molecules are removed from the tea leaves or coffee beans, often utilising supercritical fluid extraction with CO2 or standard solid-liquid extraction techniques.[3].

Clarifying agent are used to remove suspended solids from liquids by inducing flocculation (the solids begin to aggregate forming flakes, which either precipitate to the bottom or float to the surface of the liquid, and then they can be removed or collected).

Particles finer than 0.1 µm (10−7m) in water remain continuously in motion due to electrostatic charge (often negative) which causes them to repel each other. Once their electrostatic charge is neutralized by the use of a coagulant chemical, the finer particles start to collide and agglomerate (collect together) under the influence of Van der Waals forces. These larger and heavier particles are called flocs.

In winemaking, clarification and stabilization are the processes by which insoluble matter suspended in the wine is removed before bottling. This matter may include dead yeast cells (lees), bacteria, tartrates, proteins, pectins, various tannins and other phenolic compounds, as well as pieces of grape skin, pulp, stems and gums. Clarification and stabilization may involve fining, filtration, centrifugation, flotation, refrigeration, pasteurization, and/or barrel maturation and racking.



The winemaking process naturally produces sediments that can precipitate out of the wine.


Chemical structure of most common phosphatides and indication of bonds that are hydrolysed by various phospholipase enzymes.

Introduction to Degumming Degumming is the first step in the refining process to remove phospholipids, gums, waxes and other impurities from the crude oil.[52][53] The oil is treated with water or dilute acids such as phosphoric acid, which exploits the fact that the phospholipids are attracted to water because of their amphipathic nature, and turns the lipids into hydrated gums. These gums are insoluble in oil and are then separated from the oil using centrifuges. The separated gums are then dried and manufactured into emulsifying agents such as lecithin.

The Nature of Gums and Phosphatides
Crude oil obtained by screw pressing and solvent extraction of oilseeds will throw a deposit of so-called gums on storage. The chemical nature of these gums has been difficult to determine. They contain nitrogen and sugar and can start fermenting so they were at one stage thought to consist of glycolipids and proteins. Now we know that these gums consist mainly of phosphatides but also contain entrained oil and meal particles. They are formed when the oil absorbs water that causes some of the phosphatides to become hydrated and thereby oil-insoluble. Accordingly, hydrating the gums and removing the hydrated gums from the oil before storing the oil can prevent the formation of a gum deposit. This treatment is called water degumming. It is never applied to fruit oils like olive oil and palm oil since these oils have already been in contact with water during their production.

Pectinase iis an enzyme that breaks down pectin, a polysaccharide found in plant cell walls. Commonly referred to as pectic enzymes, they include pectolyase, pectozyme, and polygalacturonase, one of the most studied and widely used[citation needed] commercial pectinases. It is useful because pectin is the jelly-like matrix which helps cement plant cells together and in which other cell wall components, such as cellulose fibrils, are embedded. Therefore, pectinase enzymes are commonly used in processes involving the degradation of plant materials, such as speeding up the extraction of fruit juice from fruit, including apples and sapota. Pectinases have also been used in wine production since the 1960s.[1] The function of pectinase in brewing is twofold, first it helps break down the plant (typically fruit) material and so helps the extraction of flavours from the mash. Secondly the presence of pectin in finished wine causes a haze or slight cloudiness. Pectinase is used to break this down and so clear the wine.

They can be extracted from fungi such as Aspergillus niger. The fungus produces these enzymes to break down the middle lamella in plants so that it can extract nutrients from the plant tissues and insert fungal hyphae. If pectinase is boiled it is denatured (unfolded) making it harder to connect with the pectin at the active site, and produce as much juice.

Pectinases are also used for retting. Addition of chelating agents or pretreatment of the plant material with acid enhance the effect of the enzyme.
Pectin (from Ancient Greek: πηκτικός pēktikós, "congealed, curdled") is a structural acidic heteropolysaccharide contained in the primary cell walls of terrestrial plants. Its main component is galacturonc acid, a sugar acid derived from galactose. It was first isolated and described in 1825 by Henri Braconnot.It is produced commercially as a white to light brown powder, mainly extracted from citrus fruits, and is used in food as a gelling agent, particularly in jams and jellies. It is also used in dessert fillings, medicines, sweets, as a stabilizer in fruit juices and milk drinks, and as a source of dietary fiber.



Pectinase


A cellulase enzyme produced by Thermomonospora fusca, with cellotriose bound in the shallow groove of the catalytic domain

Cellulase is any of several enzymes produced chiefly by fungi, bacteria, and protozoans that catalyze cellulolysis, the decomposition of cellulose and of some related polysaccharides. The name is also used for any naturally occurring mixture or complex of various such enzymes, that act serially or synergistically to decompose cellulosic material.

Cellulase is used for commercial food processing in coffee. It performs hydrolysis of cellulose during drying of beans. Furthermore, cellulases are widely used in textile industry and in laundry detergents. They have also been used in the pulp and paper industry for various purposes, and they are even used for pharmaceutical applications. Cellulase is used in the fermentation of biomass into biofuels, although this process is relatively experimental at present. Medically, Cellulase is used as a treatment for phytobezoars, a form of cellulose bezoar found in the human stomach, and it has exhibited efficacy in degrading polymicrobial bacterial biofilms by hydrolyzing the β(1-4) glycosidic linkages within the structural, matrix exopolysaccharides of the extracellular polymeric substance (EPS).

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is a time-honored, state-owned enterprise in Taiwan. Its products and services have reached all corners of Taiwan to make the most trusted and well-known sugar brand. As a state-owned enterprise.
Founded in 1946 and being the leading brand of sugar production and sales in Taiwan by focusing primarily on the production and sales of sugar and sugar byproducts during its early days. TSC has n recent years, actively promoted diversification and transformation to establish several business divisions, including divisions as :
1. Sugar, 2. Biotechnology, 3. Agriculture, 4. Petroleum, 5. Livestock, 6. Leisure, and 7. Marketing Business Divisions, etc......
And has made full use of its R&D and resource advantages for promoting environmental circulation recycling and for improving social welfare, in addition to fulfilling its basic corporate responsibilities by collaborating with its notional policies and looking after peoples' livelihoods.

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TSRI is the research center of TSC in correspondence to the long-term development strategy planning. It is responsible for the research and development following government's policy and closely collaborates with different divisions to achieve the company's goals.
Most long-term research and development projects as well as highly challenging technology are also carried out by TSRI.
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Our team has experiences in sophisticated chemical and physical analysis, as well as formulation in functional food and cosmetics.
The research lab and production site are GMP, ISO, or TAF certified.
The core technologies include that are applied further in product manufacture to gain differentiated advantages within the market :
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4. Drying,
5. Formulations and deodorization, etc.

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