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http://www.novozymes.com/en/about-us/our-business/whatare-enzymes What are enzymes? Enzymes are proteins and are found everywhere in nature. The first use of enzymes occurred more than 5,000 years ago, when people stored milk in animal stomachs, which contains enzymes called “rennet,” that turn milk into cheese. This is a good example of how enzymes work as catalysts, that is, they speed up biological reactions. Novozymes’ customers use enzymes to replace chemicals in, and improve efficiency of, a wide variety of industrial processes, for example in the manufacture of margarine, beer, yogurt, concrete, leather, textiles and ethanol, where the enzymes are not part of the end-product. Enzymes are also used directly in products such as laundry detergents, where they help remove stains and enable low-temperature washing. Every enzyme has a specific function and no side effects Enzymes have one function only, and work like a key that fits in a lock. Only when the right enzyme finds the right material it can work upon, does a biochemical reaction occur. This precise correlation means you never have to worry about side effects when enzymes are added to an industrial process. For instance, when enzymes transform starch into sugar, you can be sure that is all that will happen. No other material or process will be altered or affected. Enzymes are stable, biodegradeable and environmentally friendly Enzymes work at low temperature and moderate pH and are far more stable catalysts than other chemicals or biological molecules, making them the most environmentally-friendly solution for industrial manufacturing. Enzymes are biodegradeable, and keep on working until they are dissolved, usually by other enzymes. Enzymes do not become part of the final product of the biochemical reaction which they are catalyzing. When the biochemical reaction is over, the enzyme is ready to effect the same reaction on another molecule again and again. Given the right conditions, the enzyme can go on and on for as long as needed. In some production processes, this lowers costs. When industrial enzymes leave a production plant with the waste water, the used enzymes create no hazardous waste. They do not last long in the surrounding environment as they are broken down by microorganisms found in nature. All enzymes are proteins, but not all proteins are enzymes Proteins are the building blocks of all living organisms. Humans, animals, plants and microorganisms are all made up of proteins. Every part of the human body is built of proteins. Proteins constitute about 80% of the dry weight of muscle, 70% of the dry weight of skin and 90% of the dry weight of blood. Proteins can be split into two groups: structural proteins and biologically-active proteins. Structural proteins are the main constituents of our bodies e.g. collagen, which is found in bones, tendons and ligaments, and keratin, the protein of nails, hair and feathers. Biologically-active proteins catalyze biochemical reactions in cells. These are the enzymes at the heart of Novozymes' business. What's inside an enzymes Like all other proteins, enzymes are made of amino acids. Each enzyme is made up of between a hundred and up to a million amino acids placed like pearls on a string. Each amino acid is bonded to the next by chemical bonds. Each enzyme has its own unique sequence of amino acids, which is determined by the genes in the cells. The vast majority of enzymes are made of only 20 different kinds of amino acid. The structure and function of the enzyme is determined by the order of the amino acids. Enzimlerin üç boyutlu yapıları nasıl sağlanır. In most enzymes, the string of amino acids is coiled and folded thousands of times to form a highly complex three-dimensional structure, which is unique to each enzyme. It is the chemical interactions between the amino acids that force the enzymes into their three-dimensional structure, which is held together by the many different links between the different amino acids. The unique three-dimensional structure of each enzyme determines the function of the enzyme. Even slight changes in the sequence of the amino acids on the string have a huge impact on the structure and function of the protein. With just one, or perhaps a few, amino acids replaced or switched, an enzyme may not only look different, but also act differently and convert to working on other biological molecules or treating them differently. Although enzymes are large molecules with hundreds of amino acids, only a small part of the enzyme participates in the catalysis of biochemical reactions. This is called the active site. enzyme The three-dimensional structure of the determines the appearance of the active site. The active site precisely accommodates the shape of the biological substrate (material on which it acts). The enzyme and substrate fit together like a key in a lock, and only substrates with the right shape will be transformed by the enzyme. This is what makes enzymes specific in their action. Making the perfect enzyme Enzymes have been used in food production for thousands of years - and now we make them ourselves. Enzymes in history Enzymes have been used in brewing, baking and alcohol production since prehistoric times - only our ancestors did not call them enzymes, but they certainly used them. One of the earliest written references to enzymes is found in Homer's Greek epic poems dating from about 800 BC, where mention is made of the use of enzymes in the production of cheese. The Japanese have also used naturally-occurring enzymes in the production of fermented products like sake - a Japanese schnapps brewed from rice - for more than a thousand years. Bacteria and fungi produce most industrial enzymes Naturally-occurring microorganisms are the most productive producers of enzymes. This knowledge has been exploited by industry for more than fifty years. Bacteria and fungi are the microorganisms best suited to the industrial production of enzymes. They are easy to handle, can be grown in huge tanks without light, and have a very high growth rate. Most of Novozymes' enzymes are produced by the bacterium Bacillus subtilis and the fungus Aspergillus oryzae. Both have a huge capacity for producing enzymes and are completely harmless for humans. Some microbes are more efficient enzyme producers than others The ideal microorganism grows quickly and produces lots of the desired enzyme at mild temperatures whilst consuming inexpensive nutrients. However, like most things in life, the ideal microorganism is hard to come by. Most microorganisms found in the wild are not well suited to domestication in large fermentation tanks. Some only produce tiny quantities of enzyme or take a long time to grow. Others can produce undesired by-products that would disturb industrial processes. Novozymes, on the other hand, has a large collection of microorganisms that are perfect production organisms. Combining organisms gives the perfect solution Some microorganisms have the capability of producing the perfect enzyme. Others could win the Olympic gold medal in growth and enzyme production. By combining the best from each organism, we are able to obtain a microorganism that grows very quickly on inexpensive nutrients, whilst at the same time producing large quantities of the right enzyme. This is done by identifying the gene that codes for the desired enzyme and transferring it to a production organism known to be a good enzyme producer. Microbe genes can be modified to make better enzymes Industrial enzymes need to be perfectly suited to the tasks that they perform, but sometimes the perfect enzyme for a specific job is impossible to find. This does not mean, however, that we can't make an enzyme for the job. Normally our scientists can find a naturally-occurring enzyme that is almost perfect, and using modern biotechnology we can upgrade it to the desired efficiency. This is done by altering small parts of the genes in the microorganism which codes for the production of the enzyme. These tiny alterations only alter the structure of the enzyme very slightly, but this is normally enough to make a good enzyme into a perfect enzyme. Finding and producing enzymes Learn how we select the right enzyme to do the right job by using modern biotechnology. Finding a multitude of enzymes It may be that the perfect enzyme is not in the library. If this is the case, soil samples, sometimes from very exotic locations around the world, must be examined to find the right microorganism with the capability of producing the enzyme that can remove the stain on the shirt. The starting point depends on the customer's requirement. If the enzyme needs to function under very hot conditions, our researchers in hotspring areas are the natural first port-of-call. Collecting the soil samples is the easy part, but one gram of soil contains more than 4,000 different microorganisms, each of which can produce hundreds or thousands of different enzymes! Using the biggest toolbox of screening technologies in the world, Novozymes' scientists can easily find microorganisms which are potential candidates for producing the right enzyme. This is one of the things that makes Novozymes a unique company. Selecting the right enzyme In the laboratory, our scientists start searching the many candidates collected in the wild for a microorganism which can produce the desired enzyme. Novozymes has several methods for finding the right bacterium or fungus. Normally the scientists grow the selected microorganisms in a variety of different conditions and media, narrowing down the number of suitable enzymes. The search for the right enzyme is a highly automated process. Huge robotic systems can simultaneously test a culture of microorganisms on 12 different substrates such as starch, proteins or fats. With the help of the robots, Novozymes' researchers can scan thousands of microorganisms in just a few days. The final test involves scanning the enzymes under the precise working conditions in which that enzyme will be used in the final product. The scientists can then be sure that they have found the microorganism capable of producing an enzyme to do the job. Finding the gene that makes the enzyme Having found the perfect enzyme for the job, our scientists are now ready to find the gene that instructs the microorganism to produce the enzyme. Like human beings, microorganisms are able to produce hundreds of different enzymes for solving all kinds of problem. But we are only interested in a single enzyme; the others might produce unwanted side-effects. Finding the genetic code for the right enzyme is what really sets our experts apart from normal scientists. Novozymes has the world's biggest toolbox of biotech-based screening technologies, together with some of the best scientists in the business, for finding and recreating the exact genetic code of the desired enzyme. And once the genetic code has been isolated, the researchers can improve the enzyme even further in many different ways. Transferring the gene to our production organisms Orjinal olarak bir mikroorganizmadan elde edilen bir faydalı enzimi kodlayan gen neden üretim için tasarlanmış başka bir mikroorganizmaya transfer edilir? Often the original microorganisms are not the best for producing the enzyme in large-scale production. They may not produce much enzyme or might be hard to grow in large numbers. Sometimes the original microorganism also produces a large number of other substances which might be dangerous to humans. In order to produce the enzyme in large quantities and without having to worry about the by-products, we combine the Novozymes' best own of the safe original and fast- microorganism growing with production microorganisms. The gene from the original microorganism that codes for the enzyme is inserted into the production microorganism using modern biotechnology. Our microorganisms are then able to produce exactly the same enzyme, but in larger quantities and under much safer conditions than the original microorganism. Developing the production method En etkili enzim üretimini sağlamak için hangi işlemlerin yapılması gerekir? Before we start large-scale production of the enzyme, the exact living conditions for the production organism must be determined. Even though we know our microorganisms very well, numerous tests still have to be carried out. The microorganisms feed on nutrients derived from e.g. maize (corn), soy beans, potatoes or sugars. But the right nutrients for the microorganisms depend on the enzyme which is to be produced. The same applies to the temperature, oxygen level and pH to which the microorganisms are exposed. In order to get the most efficient production, our scientists must create the right environment for the microorganisms to work to their optimum capacity. Making microorganisms produce enzymes Fermantasyon ne demektir? Ne işe yarar? Microorganisms, not larger than 1.5 micrometres, are the heart of Novozymes' business. A single bacterium or fungus is able to produce only a microscopic portion of the enzyme needed. A billion microorganisms, however, can produce the amount of enzyme that makes the business feasible. . By means of simple cell division, more and more organisms appear, all producing the enzymes which we desire. In the laboratory, a very small amount of selected microorganisms that can produce the desired enzyme is grown in a small-scale fermentation tank to get the fermentation started. Once everything is up and running, it is time to let the microorganisms grow to the best of their ability in our threestorey fermentation tanks. Everything in the tank is controlled, with nutrient quantity, temperature, pH and airflow comprising the main parameters in the process. With more than 50 years' experience, Novozymes has developed the fermentation process to perfection. Recovering enzymes from the fermentation broth Fermantasyon ortamından enzimlerin geri kazanılması nasıl gerçekleştirilir? 1. When fermentation is complete, the fermentation tank is full of one huge mix of 3microorganisms and 1unused 4valuable nutrients, 2water, enzyme. The enzymes are separated from the rest of the liquid in a large drumfilter. The filter is coated with a thick layer of wax that allows the water and enzymes to penetrate, whilst the nutrients and microorganisms are caught in the sticky surface of the wax. As the drum-filter rotates, the fermentation liquid is sprayed on. The water and enzymes are sucked into the middle of the drum, leaving the nutrients and microorganisms on the surface of the wax. A large knife then slices the wax, nutrients and microorganisms away. Following a series of other filtration processes, the enzymes are finally separated from the water using a simple evaporation process. Using surplus material as fertilizer Baktri üretim tesisilerindeki artık maddeler nasıl gübre olarak kullanılırlar. Many industries have problems with waste products. Novozymes doesn't! Dealing with nature's own technology, our waste is actually beneficial to the surrounding environment: it is used as fertilizer on local farms. Firstly, we make sure that no living or intact microorganisms leave the production plant by treating the mix of wax, nutrients and microorganisms with heat and chalk. The microorganisms in the treated fermentation waste are then ready for their final contribution to a sustainable industrial solution. The waste is used as a top-grade fertilizer on the fields of the farms surrounding the production plants, thus promoting the growth of yet another natural product, crops. Delivering the final product Once the enzymes have been formulated as either liquid or granulate, they are packed in cans or bags and are then ready to leave the factory. With factories on four continents, Novozymes is always nearby. But even when the enzyme products have left our factory Novozymes continues to work for its customers, who are often developing a brand new product which might require expert assistance. Novozymes is the number one expert on enzymes and how to handle, formulate, produce and use them. Service is therefore our second-largest product and the Tech Service department even works free of charge. How enzymes work for you Enzymes are used in a variety of the everyday products you use. And it's good for the environment as enzymes can replace chemicals and minimize energy consumption. Why we need enzymes? Even though you may not be aware of it, enzymes play a very active role in your everyday life. Saturday afternoon, washing your clothes, you may not realize that enzymes are doing some of the dirty work for you. The detergent that you are using is most likely to contain enzymes that remove the dirt and greasy stains from your clothes. Relaxing in the sun, waiting for your clothes to be cleaned, you may need something to drink. The sweet taste of the soft drink comes from syrup, which is made using enzymes. You may get hungry: A loaf of bread would come in handy. Almost all the bread you buy in the supermarket is made with a little help from enzymes. Enzymes have been used for more than 50 years in the detergent, textile, food and feed industries, to name just a few. In these industries enzymes replace chemicals and minimize water, raw material and energy consumption. Nature's own technology provides us with environmentally friendly solutions and better products. Enzymes are the natural solution to industrial problems The environment is precious to us all. But in many areas nature suffers from the heavy impact of the western way of life. All companies have a responsibility to pursue sustainable solutions to their industrial processes, and in many cases enzymes can help them do it. Chemicals used in industrial processes are one of the most severe threats to nature and man today. By using enzymes instead of chemicals, the problem is solved. Enzymes present no threat to the environment whatsoever. With enzymes we can maintain the living standards we have today and at the same time preserve the environment for our children. And enzymes do not just replace chemicals. They also reduce the consumption of raw materials, energy and water, giving real benefits to both the environment and industry. As the world's leading producer of industrial enzymes, Novozymes cannot save the planet, but we can provide some of the tools to do it. Enzymes wash your clothes at only 30oC Chemicals clean your clothes at 90°C, but what a waste! Enzymes can do the same job at only 30°C. Reducing the temperature from 90° to 30°C means great savings on energy and money. Enzymes also replace chemicals in the detergent, which means a reduction in the amount of chemical waste from both industrial and household laundry. A third benefit of putting enzymes in detergents is that they can even make your clothes look better and last longer. So what's the catch? There isn't one. If you want to take care of nature and still wear clean clothes, let enzymes do the dirty work for you. Enzymes stonewash your jeans Stonewashed jeans are the height of fashion and, as the name suggests, the traditional way of producing stonewashed jeans is to wash the jeans with stones. This is a harsh treatment both for the jeans and for the environment. The fabric of the jeans is weakened and may appear flossy, whilst the lifespan of the jeans is far shorter than that of regular blue jeans. By adding enzymes to the process there is no longer any need for stones in the wash. The look of the jeans is the same, but the. The process even saves on water, one of nature's most precious process no longer damages the fabric and the jeans therefore last much longer resources. When using enzymes to get the stonewash look, there is no need for several rinsing processes to get rid of the stones. Enzymes make good bread better The minute bread leaves the oven, the breakdown of the bread begins. It is the bread's starch content that is most "hard to please"; starch feeds on moisture, which is why bread becomes hard and unfit for consumption within a few days. By adding Novozymes' enzymes to the flour, it is possible to alter the structure of the starch in the bread so that it retains moisture better. This means that the bread remains soft for a longer period of time. Other enzymes make dough-handling much easier for the baker. Enzymes make the dough less sticky, which is a major benefit if you are making hundreds of loaves every morning. If you have ever wondered why bread from the bakers is larger and more airy, enzymes are once again the answer. Specialized enzymes can make the gluten of bread retain naturally-occurring gases that would otherwise disappear. Enzymes make your leather soft Natural, untreated leather is as stiff as metal. It therefore needs to be softened before use - and enzymes can do the job. To make leather pliable, the raw material requires an enzyme treatment called bating, which takes place before tanning. This involves dissolving and washing the protein components that stiffen the leather. The degree of bating depends on the desired properties of the finished leather. Glove leather, for example, should be very soft and pliable and is subjected to strong bating, whereas leather for the soles of shoes is only lightly bated. In the old days, dog excrement was used in the bating process, the bacteria in the excrement producing enzymes to make the leather soft. The use of enzymes in industry today is rather more hygienic. Hygiene is not the only advantage of using enzymes to treat leather products. Before leather becomes soft it undergoes several different treatments. Each treatment normally requires the use of large quantities of harsh chemicals. When removing hairs and fat from hides, enzymes can reduce the use of sulphide by 40%. Enzymes are also responsible for major reductions in the amounts of water used, as the replacement of chemicals reduces the rinsing and cleaning processes. Ultimately, a higher quality leather is achieved and the load on the environment is reduced. Enzymes make cotton look like silk Cotton treated with enzymes not only looks better, it also lasts longer. Most cotton fabrics tend to be fluffy from the minute they leave the shop. Treating the fabrics with Novozymes' unique Biopolishing enzymes removes the small hairs or fuzz that protrude from the surface of the yarn, leaving a smoother yarn surface that almost looks like silk. Biopolishing makes your clothes look brand new, even if you've washed them several times. Enzymes also play a major role in the textile industry in the desizing process. After weaving, the starch size has to be removed to prepare the fabric for the finishing steps of bleaching or dyeing. Starch-splitting enzymes are used to desize woven fabrics because of their highly efficient and specific way of desizing without harming the yarn. Enzymes make the most of fruit juice Crystal-clear juice and lots of it. This is the result of enzymes in the juice industry. The enzymes break down apple fibres, making the fruit more soluble and easier to press. Not only does this process give a much higher yield, but also a higher quality apple juice, because enzymes are able to make fruit juices completely clear. If you cut an apple in two, you will notice that the fruity part of the apple starts to turn brown. Enzymes are responsible for this process. If more enzymes are added, they speed up the process even further. By exploiting enzymes in the breakdown process, the pressing process becomes much easier. The juice becomes crystal-clear and contains all the nutrients from the apple due to the enzymatic breakdown of the fibres. In the wine industry, the same principles are used to get all the juice out of the grapes without compromising the quality of the final product. Enzymes turn corn starch into sugar syrup Sugar is the expensive element of most sweet products like candy or cola. But there is an easy way to cheaper sweets. Enzymes enable corn, cassava or wheat/potato starch to be transformed into sugar syrup. The enzymes work by rearranging and cutting up the starch molecules, turning them into liquid sugar. When the process is complete, the syrups and modified starches, which have different compositions and physical properties, can be used in a wide variety of foodstuffs, including soft drinks, confectionery, meats, baked products, ice cream, sauces, baby food, tinned fruit, preserves, and much more. Novozymes makes many specialized enzymes for the starch and sugar industries. Some of them also protect the environment. For example, one supplants the use of strong acids in the manufacture of sugar syrups. Others help manufacturers to produce products of higher quality, save energy and help ensure a safer working environment.