Method for preparing pasta under vacuum and with enzymes

Abstract

The present invention relates to the field of food production, in particular the production of pasta. The invention discloses a method for preparing pasta comprising steps of a) mixing water and a ground plant product to obtain a pasta dough, b) kneading the pasta dough, and c) forming the pasta dough into pasta of a desired shape, wherein steps b), c) and, optionally, step a) are performed in a vacuum, and wherein during step a), at least one enzyme selected from the group comprising an oxidoreductase, a carboxylester hydrolase, a hemicellulase, a glucanase, a cellulase, a transglutaminase, a protease, an amylase or a combination thereof is added to the pasta dough.

Description

[0001] Method for preparing pasta under vacuum and with enzymes

[0002] The present invention relates to the field of food production, in particular the production of pasta. The invention discloses a method for preparing pasta comprising steps of a) mixing water and a ground plant product to obtain a pasta dough, b) kneading the pasta dough, and c) forming the pasta dough into pasta of a desired shape, wherein steps b), c) and, optionally, step a) are performed in a vacuum, and wherein during step a), at least one enzyme selected from the group comprising an oxidoreductase, a carboxylester hydrolase, a hemicellulase, a glucanase, a cellulase, a transglutaminase, a protease, an amylase or a combination thereof is added to the pasta dough.

[0003] Pasta is the Italian name given to a type of food typically made from wheat flour or semolina and water. Some types of pasta may comprise further ingredients, including eggs, various spices or different types of food colourings. The production of pasta essentially comprises three steps: i) mixing the different raw materials to form a dough, ii) continuous kneading of the dough to promote the formation of an interconnected protein matrix that stabilizes the pasta and iii) forming the pasta dough into a desired pasta shape.

[0004] Industrially produced pasta is usually prepared in a dried and therefore more shelf-stable form, i.e. the freshly prepared pasta is subjected to a drying step in which the water content of the pasta is reduced by more than half. The dried pasta is then cooled to room temperature and packaged for sale.

[0005] Pasta is among the most popular foods in the world. In the Ell alone, 6.1 million tonnes of pasta, worth EUR 7.8 billion, were produced in 2022, with Italy being by far the leading EU member in pasta production (https: / / ec.europa.eu / eurostat / de / web / products-eurostat-news / w / edn- 20231025-1). Globally the pasta market is expected to undergo substantial growth, with a forecasted value of USD 24.2 billion (Global Pasta Market Overview 2024-2028, p. 4).

[0006] Traditionally, pasta is made from durum wheat (Triticum durum), which however only accounts for around 10 % of the total world wheat production. As the pasta market is continuously growing, the already scarce supply of durum wheat will continue to decrease, resulting in price increases. Fluctuations in availability, e.g. due to weather events or disruptions to global trade, have a particularly negative impact on durum wheat supply and durum wheat prices (Futures on 2 February 2024: milling wheat (mostly bread wheat) EUR 213 / t; durum wheat EUR 376 / t). It can be assumed that this problem will be further exacerbated in the future due to man-made climate change. The high price of durum wheat is prompting processors to accept batches of lower quality - and therefore at a lower price - or to use cheaper alternatives, in particular bread wheat (Triticum aestivum), which is by far the most widely cultivated type of wheat in the world. However, it is difficult to obtain a sufficiently high yield of semolina from, e.g., bread wheat. Indeed, bread wheat is typically used in the form of flour with a smaller particle size than semolina.

[0007] However, the use of low-quality durum wheat or flour from alternative wheat species significantly affects the quality of the industrially produced pasta. In particular, pasta made from, e.g., cheaper bread wheat is often very sticky after cooking and has poor firmness to the bite, characteristics that are typically associated with low pasta quality and are therefore rejected by many end consumers.

[0008] There is thus an urgent need for new and improved pasta production methods that can compensate for the above-mentioned shortcomings and quality fluctuations associated with the use of different starting materials to ensure the production of pasta of consistently high quality. This problem is solved by the present invention, especially by the subject-matter of the claims.

[0009] In a first aspect, the present invention provides a method for preparing pasta comprising steps of: a) mixing water and a ground plant product to obtain a pasta dough, b) kneading the pasta dough, and c) forming the pasta dough into pasta of a desired shape, wherein steps b) and c) and, optionally, step a) are performed in a vacuum, and wherein, during step a), at least one enzyme selected from the group comprising an oxidoreductase, a carboxylester hydrolase, a hemicellulase, a glucanase, a cellulase, a transglutaminase, a protease, an amylase and a combination thereof is added to the pasta dough.

[0010] In another aspect, the present invention provides the use of at least one enzyme selected from the group comprising an oxidoreductase, a carboxylester hydrolase, a hemicellulase, a glucanase, a cellulase, a transglutaminase, a protease, an amylase and a combination thereof for preparing pasta in a vacuum, optionally using the method of the present invention.

[0011] Pasta is the Italian designation for foodstuffs rich in starch that are prepared from doughs comprising ground plant products, in particular wheat semolina, and water. Pasta may exist both in fresh (pasta fresca) and in dried form (pasta secca). The method of the present invention is preferably used for the industrial production of dry pasta. Accordingly, the method preferably further comprises steps of: d) drying the pasta until it has a water content of less than 14 % (wt / wt), based on the total weight of the final pasta, e) cooling the dried pasta to room temperature, and f) optionally, packaging the pasta.

[0012] In some embodiments, however, the pasta produced according to the method of the invention may also be fresh pasta, which is generally prepared for consumption directly after the forming step c). In such an embodiment, the method thus does not comprise a drying step d) and, optionally, also not steps e) and f). However, fresh pasta can also be packaged so that it can be stored for a short time, usually in chilled form.

[0013] During step a) of the herein disclosed method of the invention, a relatively crumbly pasta dough is formed by mixing a ground plant product with water. The mixing ratio between the ground plant product and the water may vary dependent on the type of ground plant product that is used for preparing the pasta dough. In most scenarios, the proportion of ground plant product will however be higher than the proportion of water to obtain a sufficiently firm and workable pasta dough. If the ground plant product is flour or semolina obtained from wheat, the ground plant product is typically mixed with the water at a ratio of approximately 2: 1 to achieve a total water content in the pasta dough of more than 30 % (wt / wt), preferably of 32-34 % (wt / wt). The mixing of the ground plant product with water creates a matrix or network formed of proteins and starch present in the ground plant product, which is stabilised by intermolecular interactions. The starch granules are enclosed in the continuous protein matrix, which ensures that the pasta retains its shape during shaping and cooking. The stability of the protein network largely determines the quality of the pasta dough. Prolonged kneading of the dough during step b) promotes formation of the protein matrix and therefore increases the dough’s elasticity and formability. Kneading step b) typically takes between 1-45 min. However, the exact duration of step b) largely depends on the properties of the ground plant product used for preparing the pasta dough. It may take, e.g., about 8-12 min in case the ground plant product is T. aestivum flour or about 10-15 min when durum wheat semolina is being used. Kneading can be carried out using suitable tools known in the art, e.g., stirring rods, paddles, hooks, screws, etc. Kneading can also be done using a homogenizing mixer as disclosed in EP 1693103 A2, which is herewith incorporated by reference.

[0014] The final dough is subsequently formed into the desired pasta shape during step c). This step is frequently referred to as shaping or extruding. Typically, step c) is achieved by transferring the pasta dough into an extruder equipped with dies that determine the final shape of the pasta. The dough is pressed through these dies at comparably low pressures of about 50-150 bar, preferably 75-125 bar, such as, e.g., 100 bar, and a temperature that lies below the evaporation temperature of water to prevent an undesired foaming after the shaping of the pasta through the die. However, it is clear to a skilled person that the pasta in the method according to the invention can also be shaped in alternative ways, e.g. by mechanical rolling, pulling, blowing, punching, cutting, drawing, kneading, pressing or moulding.

[0015] The formed pasta obtained after step c) typically still has a relatively high water content of more than 30 % (wt / wt), based on the total weight of the final pasta. To prevent this fresh pasta from spoiling, it must either be prepared quickly for consumption or made more durable by reducing the water content via drying of the pasta. Drying also ensures that the pasta shape is not lost during long-term storage and facilitates transport and storage of the prepared pasta. In principle, the drying step can take place by exposing the freshly prepared pasta to the open air. However, to ensure a consistently high product quality, drying during industrial pasta production is typically carried out in a stepwise manner under highly controlled temperature, time, humidity and ventilation conditions in special zone dryers using moist, hot air. The drying of the pasta ultimately should reduce the water content of the pasta to about 10-20 % (wt / wt), preferably to less than 14 % (wt / wt), such as to about 12-13 % (wt / wt). Drying may take place at temperatures of up to 45 °C, up to 50 °C, up to 55 °C, up to 60 °C, up to 65 °C, up to 70 °C, up to 75 °C, up to 80 °C, up to 85 °C, up to 90 °C, up to 95 °C, up to 100 °C, up to 105 °C or up to 110 °C, typically, until the desired water content of the final pasta is reached. Preferably, the pasta is dried at a temperature of up to 90 °C. To avoid the development of cracks in the pasta, drying should not be performed too fast. On the other hand, too slow drying may favor the growth of mold. Preferably the pasta should be dried for about 200-400 min, preferably for 250-350 min, more preferably for 275-325 min, e.g., about 300 min. Drying can be performed by any suitable method known in the state of the art, i.e. the pasta may first be pre-dried at room temperature before most of the moisture is removed in a final drying step. The final drying step may first comprise a step wherein the pasta is exposed to high temperatures and humidity, before in the second step, the temperature is slowly reduced and cold air is provided to stabilize the final pasta. An exemplary method for drying pasta is disclosed in EP 2416669 A1 , which is incorporated herewith by reference.

[0016] In an optional step f), the pasta may be packaged to facilitate transport and distribution of the pasta. To ensure maximum shelf-life, the pasta may be packaged in a vacuum-sealed package, e.g. a bag or a blister package. Optionally, the pasta is packaged in ecologically friendly, biodegradable package, e.g. a bag made of paper, or a bag of bio-degradable plastic.

[0017] The plant product used for the production of pasta using the method of the invention may be any of, e.g., wheat, rye, oat, barley, millet, rice, maize, buckwheat, potatoes, soy, quinoa, lentils, beans, peas, chickpeas, lupins and cassava. It is typically derived from an edible part of the plant, often, the seeds. However, the ground plant product can also be further processed before use, e.g., by extracting non-desired components. For example, if the lupins are not sweet lupins, alkaloids can be extracted by saltwater soaking. Plant products from different sources can also be mixed. Preferably, the plant product is a cereal grain, e.g., from wheat, rice, rye, oats, barley, millet, and maize. “Cereal” refers to a grass cultivated for its edible grain, which is the dry fruit (caryopsis) of the cereal plant. Rice or maize flours as well as flours made from legumes such as lentils or beans are becoming increasingly popular as a gluten-free alternatives to wheat products. Thus, in one embodiment, the ground plant product is gluten-free.

[0018] However, most preferably, the ground plant product that is used for forming the pasta dough is ground wheat. The wheat may be any type of wheat, e.g., it may be Triticum aestivum, Triticum durum, Triticum dicoccum, Triticum spelta, Triticum monococcum or a mixture thereof. However, preferably, the wheat used in the method of the invention is Triticum aestivum.

[0019] Triticum aestivum, better known as common wheat or bread wheat, constitutes approximately 90 % of the globally produced wheat. T. aestivum is occasionally also differentiated into soft wheat and hard wheat. Soft wheat refers to T. aestivum with a lower gluten content than hard wheat. T. aestivum is a hexapioid wheat species and contains on average about 11 % protein (from 9 to 12%) (dry weight) (Zilic et al., 2011). Gluten constitutes about 75-85 % of said total grain proteins and stores carbon, nitrogen and sulphur to support seed germination. Gluten is a protein mixture composed of prolamins and glutelines, which, in the context of wheat, are usually referred to as gliadins and glutenins, respectively. Glutenins form protein aggregates stabilized via intermolec- ular disulfide bonds that become attached to the monomeric gliadins. Collectively, gluten proteins form a matrix with viscoelastic and adhesive properties (Shewry et al., 2002). T. aestivum carries rather softer grains than durum wheat that can be conveniently milled into fine flour. As a result, the gluten networks become exposed and can be easily brought together during the mixing and kneading of dough made of bread wheat, thereby creating new protein-protein interactions within the growing gluten network. Accordingly, doughs prepared from T. aestivum are usually high in strength and elasticity and are therefore preferred for bread making.

[0020] T. aestivum can also be used for products other than bread, e.g., for cakes, cookies, biscuits, or wafers. For these applications, flour from softer T. aestivum variants is preferred. T. aestivum of intermediate hardness is preferred for the preparation of flour that can be used in laminated crackers and hard biscuits.

[0021] In principle, flour from bread wheat may also serve, and is indeed frequently used, as raw material for pasta production. However, the high elasticity and stickiness of moistened bread wheat flour tends to negatively affect the production of pasta, resulting in a final product that tends to be comparably sticky and soft after cooking. If flour from soft T. aestivum wheat is used, the final pasta will be even stickier and softer.

[0022] Therefore, the wheat species used for preparing pasta traditionally is Triticum durum, herein also referred to as durum wheat. T. durum is a tetrapioid wheat species, derived from the tetrapioid species Triticum dicoccum, and the second most cultivated species of wheat after T. aestivum. The Latin name Durum translates as hard, as the grains of T. durum are known for being highly resistant to milling. T. durum is therefore mostly processed into more coarse-grained semolina rather than into fine flour. T. durum is described to contain slightly higher levels of gluten compared to T. aestivum (Zilic et al., 2011). However, because the gluten- and starch-rich endosperms are often only partly cracked in Durum semolina, the gluten is less readily available. Doughs made from durum wheat semolina are therefore less elastic and sticky than those made from bread wheat flour. Accordingly, semolina made from T. durum is only rarely used for baking bread and is instead particularly suitable for making pasta. However, the comparatively high procurement costs and limited availability of durum wheat force many pasta manufacturers to use the cheaper wheat T. aestivum or durum wheat of lower quality, which can have a negative impact on the processability of the milled product.

[0023] Triticum spelta, also referred to as spelt or dinkel wheat, is a relict crop, which enjoys growing popularity among more health-oriented consumers. Spelt is a hexapioid wheat and is most commonly used for baking breads, rolls and other pastries, as it shares many properties with T. aestivum. It may also be employed in beer brewing or for the distillation of spirits.

[0024] Triticum dicoccum, better known as emmer wheat or hulled wheat, is considered to be one of the oldest domesticated crop species. Emmer is a particular stress-resistant wheat and therefore can grow on comparably poor soils. Similar to T. durum, emmer possesses relatively hard grains. Emmer wheat is most famously employed in the production of a particular type of Italian bread (pane di farro) but may also be used for garnishing soups or for preparing beer.

[0025] Similar to emmer and spelt, Triticum monococcum (einkorn wheat) is an ancient wheat and is characterized by a high protein and fat content. It is commonly consumed in Provence, France, and may be used as an ingredient of bulgur.

[0026] In the present invention, the preferred wheat species used for preparing pasta is Triticum aestivum, because it is by far the most cultivated wheat species in the world and therefore considerably cheaper and far more available than T. durum, T. dicoccum, T. spelta and T. monococcum. Accordingly, using T. aestivum for producing the pasta thus reduces production costs and results in a final product that may be sold at a lower price.

[0027] The plant product may also be a combination of different wheat species, e.g., it may be a mixture of T. aestivum and T. durum. Preferably, the pasta dough formed in step a) of the method according to the invention comprises more T. aestivum than T. durum. The percentage of T. aestivum in the pasta dough may be, e.g., 0-100 %, e.g., 90-100 %, 80-90 %, 70-80 %, 60-70 %, 50-60 %, with T. durum ad 100 %. % refers to a weight per weight ratio (wt / wt) unless otherwise mentioned. In other embodiments, the pasta dough may however also comprise more T. durum than T. aes- tivum, i.e., the percentage of T. aestivum being 40-50 %, 30-20 %, 20-30 %, 10-20 % or 0-10 %, with T. durum ad 100 %. It may also comprise only T. durum and no T. aestivum.

[0028] The pasta dough may however also be formed using one or more of T. dicoccum, T. spelta and / or T. monococcum, optionally, in a mixture with T. aestivum. The pasta dough may therefore also have a T. dicoccum content of, e.g., 0-100%, e.g., 90-100 %, 80-89 %, 70-80 %, 60-70 %, 50- 60 %, 40-50 %, 30-40 %, 20-30 %, 10-20 % or 0-10 %, with any other of the herein described wheat species, e.g., T. aestivum, ad 100 %. The pasta dough may therefore also have a T. spelta content of, e.g., 0-100%, e.g., 90-100 %, 80-89 %, 70-80 %, 60-70 %, 50-60 %, 40-50 %, 30- 40 %, 20-30 %, 10-20 % or 0-10 %, with any other of the herein described wheat species, e.g., T. aestivum, ad 100 %. The pasta dough may also have a T. monococcum content of, e.g., 0- 100%, e.g., 90-100 %, 80-89 %, 70-80 %, 60-70 %, 50-60 %, 40-50 %, 30-40 %, 20-30 %, IQ- 20 % or 0-10 %, with any other of the herein described wheat species, e.g., T. aestivum, ad 100 %.

[0029] In the context of the invention, the term “ground plant product” is used as a generic term encompassing flour, semolina or a mixture thereof.

[0030] “Flour” refers to a fine powder obtainable by grinding or milling raw plant products such as cereal grains to a particle size of preferably less than 150 pm, e.g. of 50-149 pm. Smaller particles may also be contained. Cereal flour may either be whole grain, i.e., it may be prepared from the endosperm, germ and bran together, or it may be a refined flour, i.e., it may be prepared only or partly from the starch-rich endosperm.

[0031] Semolina refers to the more coarse, purified millings of various cereals including wheat, rice or maize. It is however typically obtained from milling hard wheat species, e.g. T. durum. The average particle size of semolina and flour may vary considerably depending on the employed milling technique and the type of cereal used. However, the particle size of semolina is commonly in the range of more than 250 pm to about 1000 pm, e.g. from about 300 pm to about 750 pm (en.wik- ipedia.org / wiki / Semolina; Sacchetti et al., 2011) and is therefore largerthan that of flour. However, ground grain products with particle sizes smaller than 250 pm are often still referred to as semolina. In the context of the invention, the ground cereal grain is therefore considered to be semolina if it has a particle size of more than 150 pm, e.g. of 150-1000 pm. During semolina production from wheat, the bran and germ of the wheat are flaked-off, while the starch-rich endosperm is cracked into coarse fragments. These endosperm pieces form the actual semolina when separated from the bran. The semolina can optionally be further ground into finer particles to produce flour. In the context of the invention, the mean particle size is preferably analysed by sieving using, e.g., a test sieve shaker such as a Ro-Tap® (www.haverparticleanalysis.com / en / sieve-analysis / ro-tapr- test-sieve-shaker / ), Vibratory Sieve Shaker AS 200 Control (www.retsch.com / products / siev- ing / sieve-shakers / as-200-control / function-features / ). An air jet sieve such as the Laboratory Airjet Lab sieve KLS (gkm-net.de / en / laboratory-air-jet-lab-sieves.html) can also be used. Alternatively, particle size may also be determined by laser diffraction or spectrometry (Hareland, 1994).

[0032] In one embodiment, the plant product, e.g., the wheat used in the method of the invention is milled to flour with an average particle size of less than 150 pm, more preferably less than 125 pm, less than 110 pm or less than 100 pm. The finer the particle size of the ground wheat, the larger the reaction surface for the at least one enzyme added to the pasta ingredients during step a) of the method of the invention. E.g., at least 50 %, preferably at least 75 %, at least 80 %, at least 85 %, at least 90 %, at least 95 %, at least 96 %, at least 97 %, at least 98 % or at least 99 % of the ground plant product is provided as flour e.g., as defined herein. Optionally, 100 % of the ground plant product used for preparing the pasta is milled to flour. Accordingly, typically, less than 50 %, preferably, less than 25 %, less than 20 %, less than 15 %, less than 10 %, less than 5 %, less than 4 %, less than 3 %, less than 2 % or less than 1 % of the ground plant product are provided as semolina. Optionally, the ground plant product used for preparing the pasta does not comprise any semolina at all.

[0033] The proportion of semolina may however also be higher, e.g., at least 25 %. It may for instance constitute at least 30 %, at least 35 %, at least 40 %, at least 45 % or at least 50 % of the ground plant product used for preparing the pasta. In preferred embodiments, the majority of the ground plant product used for preparing the pasta may be provided as semolina, e.g. at least 60 %, at least 70 %, at least 80 %, at least 90 %, at least 95 %, at last 95 %, at least 96 %, at least 97 %, at least 98 % or at least 99 %. Most preferably, the entire ground plant product for preparing pasta is provided as semolina. Preferably, the semolina used for the preparation of pasta has an average particle size larger than 150 pm, preferably larger than 200 pm or larger than 300 pm, more preferably, larger than 400 pm, larger than 450 pm, or larger than 500 pm.

[0034] Depending on the specific pasta recipe, further ingredients in addition to the ground plant product and water may optionally be mixed to form the pasta dough such as, e.g., eggs, vegetables, spices or food colorings. In principle, eggs may also be used instead of plain water as a source of liquid. However, since the method of the present invention preferably is for preparing dried pasta suitable for long-term storage, the pasta dough preferably does not comprise eggs. The food coloring may be an edible colour additive, e.g. a food colour such as betanin, anthocyanin, beta-carotene, riboflavin, tartrazine, curcumin, indigo carmine, Allura Red AC, sepia, xanthin, xe- axanthin, lutein or Quinoline Yellow WS, or an extract from a plant, such as a spinach extract, a red beet extract, a carrot extract, or Calendula officinalis extract. Yellowish or reddish food colourings such as riboflavin, curcumin or beta-carotene are often used to give pasta made from comparatively colourless flours such as bread wheat flour an appearance similar to that of pasta made from durum wheat. Egg yolk can also be used for this purpose. Additional optional ingredients may be, e.g., a flavouring, e.g., sugar or sweetener.

[0035] Upon mixing of the ingredients during step a), the water is absorbed by the ground plant product, i.e., the flour and / or semolina, especially by starch and gluten proteins present in the ground plant product.

[0036] In the method of the invention, steps b), c) and, optionally, step a) are performed in a vacuum. Preferably, all steps a)-c) are performed in a vacuum. In the latter case, the method of the invention is herein also referred to as a “total” or “full vacuum” pasta production process. In contrast, the method of the invention is considered to be only a “partial vacuum” pasta production process when only steps b) and c), i.e., not step a), are performed in a vacuum.

[0037] In theory, the term “vacuum” refers to a space devoid of any matter. However, it is understood that a perfect vacuum, i.e., a space that contains no particles whatsoever, cannot be created and maintained in practice. When the term “vacuum” is used in the context of the invention, it therefore refers merely to an approximation to such a perfect vacuum. Accordingly, the term “vacuum” is herein defined to relate to a closed space or system with a gas pressure that is significantly lower than the local atmospheric pressure. Atmospheric pressure refers to the pressure within the atmosphere of Earth. The standard atmosphere (symbol: atm) is a unit of pressure defined as 101 ,325 Pa (1 ,013.25 hPa), which is equivalent to 1 ,013.25 millibars (https: / / en. Wikipedia. org / wiki / Atmospheric_pressure). However, a person skilled in the art will be aware that the atmospheric pressure on Earth slightly varies with the altitude and weather.

[0038] A vacuum may be measured, e.g., with a differential pressure transducer that has one port open to atmosphere. Gas pressure that is measured relative to atmospheric pressure is also referred to as gauge pressure. Gauge pressure is positive for pressures above atmospheric pressure, and negative for pressures below it. A vacuum is therefore characterized by a negative gauge pressure. Thus, in the context of the present invention, the phrase “steps b), c) and, optionally, step a) are performed in a vacuum” is intended to mean that steps b), c) and, optionally, step a) are to be performed at a negative gauge pressure. Preferably, steps b), c) and, optionally, step a) are performed at a negative gauge pressure of less than -500 hPa, preferably of less than -600 hPa, less than -700 hPa or, more preferably, of less than -800 hPa. In a particularly preferred embodiment, steps b), c) and, optionally, step a) are performed at a gauge pressure of less than -900 hPa, e.g., of -980 hPa or even lower. At a vacuum level of -980 hPa, the oxygen partial pressure is only approx. 2% of the oxygen partial pressure of the atmospheric ambient pressure. Preferably, steps b), c) and, optionally, step a) may all be performed at the same gauge pressure, e.g., they may all be performed at a gauge pressure of -980 hPa. In alternative embodiments, steps b), c) and, optionally, step a) may also be performed at different negative gauge pressure, e.g., step b) may be performed at a lower gauge pressure than step c) or the other way around.

[0039] Many mixers (kneaders) used during industrial pasta production nowadays work under vacuum. Negative gauge pressures of less than -800 hPa, e.g., -980 hPA, can be generated, e.g., in total vacuum systems developed by the Italian company Fava S.p.A.. The preparation and kneading of pasta doughs in vacuum is associated with various advantages. For instance, mixing and kneading of the pasta dough in a vacuum avoids the development of air bubbles in the pasta dough that would otherwise refract the light and make the pasta appear opaque (Kruger et al., 1998, p. 30). The introduction of air bubbles into the pasta dough may further negatively affect the stability of the final pasta structure. Moreover, vacuum applied during pasta dough mixing and kneading has been found to prevent a fading of the natural colour pigments present in the flour / semolina, as, in the absence of oxygen, enzymes that are naturally present in the ground plant product and are capable of catalysing the oxidation and destruction of flour / semolina pigments such as carotenoids and flavonoids are inactive due to the lack of oxygen.

[0040] Step a) of the method of the invention is further characterized in that at least one enzyme selected from the group comprising an oxidoreductase, a carboxylester hydrolase, a hemicellulase, a glu- canase, a cellulase, a transglutaminase, a protease, an amylase and any combination thereof is added to the pasta dough during mixing.

[0041] It has been a long-standing goal to strengthen the protein structure and thus optimize the quality of pasta made from ground plant products, in particular pasta made from flour or semolina that is not derived from durum wheat. For instance, the use of structure-building and cross-linking enzymes has been contemplated in the past to improve the quality of pasta produced from flour of the cheaper bread wheat T. aestivum. In the context of the invention, the “quality” of pasta refers to the presence or absence of at least one characteristic, in particular of cooked pasta, that is considered particularly desirable by the end consumer. Characteristics that indicate high pasta quality include, for example, increased firmness to the bite of the cooked pasta, which also correlates with increased cooking tolerance. Cooking tolerance refers to the ability of pasta to maintain a high level of firmness even after prolonged cooking times, e.g. cooking times that exceed the optimal cooking times recommended by the pasta manufacturer. Another characteristic associated with a high pasta quality is a reduced stickiness of the cooked pasta.

[0042] The firmness and stickiness of cooked pasta depends on the integrity of the pasta matrix. The term “pasta matrix” is intended herein to relate to the structural network of the pasta, which is mainly formed by proteins such as gluten as well as the starch present in the ground plant product as described herein. The pasta matrix is considered strong if, e.g., the structural protein network of the pasta is very dense, i.e. when it comprises multiple cross-links between individual proteins or protein aggregates. Heating hydrated gluten above 55 °C during cooking of pasta leads to protein coagulation through irreversible protein-protein interactions, which also stabilises the structure and gives the cooked pasta the desired texture when eaten (Bocket al., 2015). However, during cooking, the starch present in the pasta matrix begins to gelatinise. The starch acts not only as a filler in the continuous protein matrix of the dough, but also appears to form a bi-contin- uous network with proteins (Horstmann et al., 2017). When dried pasta is brought into contact with water during cooking, it absorbs water and starts to swell. In addition, intermolecular bonds of starch molecules will break down, the starch dissolves and is progressively washed out of the pasta matrix. This process will be accelerated by high temperatures. The speed of this starch gelatinisation in spaghetti samples further depends on the protein content and is lower at low protein concentrations (Grzybowski et al., 1977). The more stable the protein structure, the less starch is released during cooking (Dexter et al., 1983; Cunin et al., 1995). The amount of starch released during cooking, also known as cooking loss, determines the stickiness of the pasta. Furthermore, with a stable protein network, the later onset of starch gelatinisation increases the firmness and cooking tolerance of the pasta. Due to the lower protein content and poorer gluten quality, soft wheat flours form a weaker gluten structure compared to durum wheat flour / semolina. In addition, the higher starch content in soft wheat flours means that the loss of starch grains from the already weaker protein matrix is greater, making pasta made from soft wheat stickier than pasta made from durum wheat.

[0043] By adding flour treatment agents such as enzymes, it is possible to significantly improve the quality of pasta made from e.g., bread wheat flour, but also of durum wheat semolina.

[0044] This is why, in pasta production, enzymes such as hemicellulases, lipases and oxidases have previously been successfully used to specifically control and / or manipulate the properties and the processability of the pasta dough and to compensate for quality fluctuations of the used raw materials (Menzi, 1967; Qi Si and Drost-Lustenberger, 2002; Brijs et al., 2004; Popper, 2020; Popper et al., 2013).

[0045] Indeed, the process of producing pasta normally offers optimal conditions for the use of such enzymes. Pasta is produced by preparing a dough typically using lukewarm water (30-40 °C) in which the water forms a continuous phase that allows for diffusion processes. The dough is then moulded before being dried with moist, hot air. The cooling limit temperature - i.e., the temperature of the pasta during drying while there is still free water - remains below 50 °C for a sufficiently long time, giving enzymes enough time to influence the final pasta product. However, up until now, it has been assumed that the use of vacuum during pasta production prevents the efficient utilisation of enzymes, especially those enzymes that require oxygen for their catalytic function. Indeed, the simultaneous use of vacuum and enzymes in pasta production would have been considered to be mutually exclusive: either one benefits from the numerous advantages associated with the application of a vacuum during pasta production but dispenses with the use of oxygen-dependent enzymes in particular, or one utilises the catalytic activity of these enzymes, but must forego the use of vacuum.

[0046] Accordingly, a noticeable activity in particular of oxygen-dependent enzymes during pasta preparation was not to be expected in a partial or full vacuum pasta production method. Contrary to this expectation, the present inventors were however able to show that the addition of enzymes to the pasta dough, including oxygen-dependent oxidoreductases, can have a positive effect on the quality of the final pasta even if steps a)-c) of the process according to the invention are carried out under vacuum, in particular, at a negative gauge pressure of less than -800 hPa or even of -980 hPa. This effect is particularly noticeable when T. aestivum flour or semolina is used to make the pasta, which, as described above, normally tends to form highly elastic and sticky doughs after the addition of water that are not optimal for pasta production. However, enzymatic effects in vacuum are also observable when other ground plant products, e.g., flours and semolina from other types of wheat are used. The use of the herein disclosed enzymes can therefore also compensate for fluctuations in the quality of the starting material used for preparing pasta.

[0047] Accordingly in another aspect, the present invention also provides the use of at least one of the herein described enzymes selected from the group comprising an oxidoreductase, a carboxylester hydrolase, a hemicellulase, a glucanase, a cellulase, a transglutaminase, a protease, an amylase and a combination thereof for increasing the firmness and cooking tolerance and / or decreasing the stickiness of cooked pasta prepared in a vacuum, optionally using the method of the present invention.

[0048] “Increasing the firmness and cooking tolerance of cooked pasta” as used herein is intended to mean that the firmness and cooking tolerance of cooked pasta as defined herein is stronger / higher, when during production of said pasta using the method of the invention, at least one of the herein disclosed enzymes is added during step a), compared to the firmness and cooking tolerance of cooked pasta that has been produced without adding any enzymes. Likewise, “decreasing the stickiness of cooked pasta” is intended to mean that the stickiness of cooked pasta is less noticeable when, during production of the pasta according to the method of the present invention, at least one of the herein disclosed enzymes is added during step a), compared to the stickiness of cooked pasta that has been produced without adding any enzymes. The firmness and / or stickiness of cooked pasta can be tested as described in the example below.

[0049] In the context of the invention, addition of at least one of the herein disclosed enzymes to the pasta dough during step a) of the method according to the invention may increase the firmness and / or reduce the stickiness of cooked pasta by at least 5 %, preferably at least 10 %, at least 15 %, at least 20 %, at least 25 %, at least 30 %, at least 35 %, at least 40 %, at least 45 % or at least 50 %.

[0050] Most surprisingly, it was found that despite the lack of oxygen in the method of the invention, the use of, e.g., oxidoreductases such as glucose oxidase (GOX) noticeably reduced the stickiness of the cooked pasta compared to a pasta sample that was obtained by the same method but without the addition of enzymes.

[0051] Therefore, in a preferred embodiment, the at least one enzyme that is added in step a) of the method of the present invention is an oxidoreductase. Oxidoreductases are enzymes that catalyse the transfer of electrons from an electron donor to an electron acceptor while utilizing NADP+ or NAD+ as cofactors. Preferably, the oxidoreductase is a hexose oxidase (EC 1.1.3.5), i.e., an enzyme capable of catalysing the transformation of mono- and oligosaccharides such as beta-D- glucose, D-galactose, xylose, arabinose, cellobiose, lactose, maltose, maltotriose or malto- tetraose to corresponding lactones. Most preferably, the oxidoreductase is a glucose oxidase (EC 1.1.3.4). Glucose oxidases suitable for use in the method of the invention are typically derived from Aspergillus niger or Penicillium chrysogenum. However, the glucose oxidase may also be isolated from bacteria, red algae and from the saliva of honeybees (Stellmach, 1988).

[0052] In the presence of oxygen, glucose oxidase (GOX) catalyses the oxidation of p-D-glucose to D- gluconic acid and hydrogen peroxide (H2O2) (Stellmach, 1988). The simplified reaction equation of D-glucose conversion by GOX is as follows:

[0053] GOX

[0054] P-D-glucose + O2 > > D-gluconic acid + H2O2

[0055] Glucose oxidase is frequently used during bread-making because its catalytic activity leads to an increase in elasticity and stability of the bread dough, which can, e.g., increase the bread volume due to an increased gas holding capacity. The use of glucose oxidase in baked goods has been documented since 1957 (Luther, 1957).

[0056] The mechanism for the effect of glucose oxidase in wheat dough is not yet fully understood at the molecular level (Hanft et al., 2006). However, the positive effects on dough quality are usually attributed to reactions that involve the hydrogen peroxide formed within the dough matrix. The H2O2 has a non-specific effect and can be involved in various reactions as a strong oxidising agent. These reactions include the oxidation of the free thiol groups of soluble and insoluble proteins in flour (Vemulapelli et al., 1998; Rasiah et al., 2005), the formation of dityrosine bonds in glutenin (Takasaki et al., 2005; Pescador-Piedra et al., 2010) and the oxidative gelation of water- soluble pentosans (Rasiah et al., 2005; Takasaki et al., 2005). The oxidation of the free thiol groups in the insoluble protein fractions is said to be considerably stronger than the oxidation of the soluble proteins (Vemulapelli et al., 1998; Rasiah et al., 2005). The insoluble protein fractions are therefore more susceptible to oxidation reactions.

[0057] Bonet et al. (2006) investigated the effect of glucose oxidase on gliadins and glutenins and were able to prove by means of cryo-electron microscopy (cryo-SEM) that a more homogeneous and finer protein network is formed by the addition of glucose oxidase, which was attributed to the stronger linking of the network by covalent disulphide bridges. The effect of oxidative gelling on the quality of wheat doughs is evaluated differently. For example, the binding of free water during the H2O2-induced gelation of the pentosans could explain the drying effect observed on the surface of wheat doughs when glucose oxidase is added (Miller et al. 1999; Courtin et al., 2002). However, other sources also show that oxidative gelling hinders the formation of the gluten network, as there is competition for the free water and the movement of the gluten fractions is limited by the formation of the gel (Wang et al., 2002; Dagdelen et al., 2007).

[0058] Previous studies have primarily focussed on the effect of glucose oxidase in bread doughs. Due to the different dough properties of bread and pasta doughs, mainly caused by the different water content, the findings on the effect of glucose oxidase in bread doughs (water content approx. 60 %) cannot necessarily be transferred to the application in pasta (water content 30-33 %). Due to the higher water content, the conditions for the formation of the gluten network in baked goods are significantly better. Furthermore, the production process for baked goods is better suited to the utilisation of enzymes than the production process for pasta due to the fermentation times.

[0059] Given that the above-described catalytic activity of glucose oxidase depends on oxygen, it was completely unexpected that the use of this enzyme in the method of the present invention would result in any observable advantageous effect on the quality of the finished and cooked pasta. It seems that even when prepared and processed in a full vacuum environment, i.e., in a method of the invention in which all of steps a), b) and c) are performed in a vacuum, the dough may, for a short period of time, retain sufficient oxygen for the glucose oxidase to become catalytically active. Without being bound by theory, one contributing factor could be that the individual flour particles are hydrated faster and more thoroughly without atmospheric air, which makes it easier for the glucose oxidase to reach its substrate during dough production and thus positively influence the dough properties. An exemplary glucose oxidase suitable for use in the method of the invention may comprise an amino acid sequence having SEQ ID NO: 6 or SEQ ID NO: 7. An enzyme of SEQ ID NO: 6 is commercially available as “Sternzym Gloxy”, and of SEQ ID NO: 7 as “Sternzym Gloxy TGO”, both from SternEnzym GmbH & Co. KG, Germany. The enzyme may also comprise an amino acid sequence having at least 80 %, at least 85 %, at least 90 %, at least 95 %, at least 96 %, at least 97 %, at least 98 %, or at least 99 % sequence identity to either SEQ ID NO: 6 or SEQ ID NO: 7. The enzyme may also consist of SEQ ID NO: 6 or 7.

[0060] The at least one enzyme added during step a) may also be another type of oxidoreductase that normally requires the presence of oxygen for its catalytic activity. For instance, in one embodiment, the enzyme may be, e.g., a sulfhydryl oxidase which utilizes molecular oxygen as electron acceptor to oxidize free thiol groups in proteins, thereby promoting the formation of disulfide bonds.

[0061] For instance, the enzyme of the invention, e.g., the oxidoreductase, may also be a sulfhydryl oxidase comprising an amino acid sequence having SEQ ID NO: 3. Such an enzyme is commercially available as "Thiolase" from SternEnzym GmbH & Co. KG. The enzyme may also comprise an amino acid sequence having at least 80 %, at least 85 %, at least 90 %, at least 95 %, at least 96 %, at least 97 %, at least 98 %, or at least 99 % sequence identity to SEQ ID NO: 3, wherein the enzyme is also capable of strengthening the protein matrix in aggregates formed by ground plant product grains, in particular via catalysing the formation of disulphide crosslinks, e.g., as defined herein. The enzyme may also consist of SEQ ID NO: 3.

[0062] In another embodiment, the oxidoreductase may also be a phenol oxidase, in particular a laccase (EC 1.10.3.2). Laccases are capable of oxidizing a large variety of aromatic compounds. The reaction products of laccases often continue to react non-enzymatically; accordingly, laccase favours the generation of polymers and has the potential to crosslink food polymers such as proteins and non-starch polysaccharides, such as ferulic acid-substituted arabinoxylan hemicellulose, which results in arabinoxylan network formation. In addition, laccase may oxidize the tyrosyl residues of gluten proteins or enhance the disulphide bridge formation in gluten polymers via ferulic acid-derived radicals. In consequence, protein aggregation is increased (Selinheimo, 2008).

[0063] Therefore, the enzyme of the invention may also comprise an amino acid sequence having SEQ ID NO: 5. Such an enzyme is commercially available as “Suberase” from Novozymes A / S. The enzyme may also comprise an amino acid sequence having at least 80 %, at least 85 %, at least 90 %, at least 95 %, at least 96 %, at least 97 %, at least 98 %, or at least 99 % sequence identity to SEQ ID NO: 5, wherein the enzyme is also capable of strengthening the protein matrix in aggregates formed by ground plant product grains and catalysing the formation of an arabinoxylan network, e.g., as defined herein. The enzyme may also consist of SEQ ID NO: 5. The oxidoreductase may also be a peroxidase (EC 1.1.11.7). Previous studies have demonstrated that the use of peroxidases can significantly increase the hardness and decrease in adhesiveness of doughs formed from wheat flour (Revanappa et al., 2014). Without being bound by theory, it is assumed that peroxidases may catalyse the formation of cross-linking between arab- inoxylans as well as protein-arabinoxylan that could be responsible for the alteration of the whole wheat flour dough characteristics.

[0064] The oxidoreductase may also be a tyrosinase (EC 1.14.18.1) isolated, e.g., from Trichoderma reesei. Tyrosinase can catalyse the oxidation of tyrosine side chains in proteins which can contribute to the formation of crosslinks between wheat gliadins. Accordingly, tyrosinase is capable of increasing dough strength.

[0065] In a further embodiment, the oxidoreductase may also be a lipoxygenase, which is capable of converting free unsaturated fatty acids present in the dough into peroxides that, as described above, may contribute to the oxidation of free thiol groups of soluble and insoluble proteins in flour, thereby strengthening the protein network of the dough.

[0066] The oxidoreductase may also be an amino acid oxidase, which is capable of converting L-amino acids into an a-keto acid (2-oxo acid), ammonia, and hydrogen peroxide.

[0067] In another embodiment, the at least one enzyme added during step a) of the method of the invention may also be hydrolytic enzyme, such as, e.g., a carboxylester hydrolase. Carboxylester hydrolases (also known as carboxylic ester hydrolases) are enzymes that catalyse the hydrolysis of carboxylic esters into alcohols and carboxylic acids. Representatives of this class of enzymes are lipases such as, e.g., triacylglycerol lipases (EC 3.1.1.3), which catalyse the hydrolysis of fats (lipids) into their basic components, fatty acids and glycerol. Other carboxylester hydrolases that may be added during step a) of the method of the invention include phospholipases (A1 , A2, B, C or D) or a galactolipase.

[0068] Besides protein and starch, cereals such as wheat comprise a variety of lipids. The majority of lipids in wheat are esters formed by glycerol and fatty acids. These so-called glycerolipids may be triglycerides, diglycerides or monoglycerides, mono- and di-galactosyl-diglycerides, phospholipids such as / V-acyl-phosphatidyl-ethanolamine, phosphatidyl-ethanolamine, phosphatidyl-glyc- erol, or phosphatidyl-choline. Other lipids that can be found in wheat are free fatty acids as well as sterol-based lipids and glycol-sphingolipids (Morrison, 1994). Without intending to be bound by the theory, lipases, e.g., triacylglycerol lipases may interact with the acylglycerides present in the ground plant product, e.g., the ground cereal grains and catalyse their hydrolysis into fatty acids as well as partial glycerides, i.e. , mono- and diglycerides. These partial glycerides act as emulsifiers, i.e., they possess a polar or hydrophilic part and a non-polar or hydrophobic part. Phospholipases and galactolipases likewise split off fatty acids, which are highly hydrophobic, from the corresponding polar lipids, hence increasing the polarity of the remaining phospholipid or galactolipid. The resulting lyso-lipids (partial glycerides) may subsequently interact with the starch. Furthermore, fatty acids also tend to react with the hydrophobic regions of helical starch molecules, altering their pasting properties (Kibar et al., 2014). In addition, partial glycerides may also interact with gluten to promote aggregation and crosslinking of the protein matrix, which further prevents the release of starch when the ground grains are brought in contact with water. Finally, the fatty acids released upon hydrolysis of lipids may serve as a substrate for lipoxygenases that are naturally present in the ground plant products, and which are able to convert the fatty acids into peroxides. Like hydrogen peroxide described above, these peroxides can have a strong oxidising effect and thus contribute to the oxidation of free thiol groups of soluble and insoluble proteins in flour.

[0069] As can be seen from the examples below, the use of a triacylglycerol lipase under full vacuum in the method according to the invention increased the firmness of the cooked pasta, which was particularly noticeable when the pasta was subjected to longer cooking times. The latter observation is indicative of the ability of the triacylglycerol lipase to increase the cooking resistance of pasta. The activity of carboxylester hydrolases does not require the presence of oxygen. The effects associated with the use of lipase in vacuum can therefore possibly also be explained by the fact that the enzyme can reach its substrates better in a vacuum environment. However, the observed result associated with use of lipase in a full vacuum pasta production process is still surprising given that, e.g., the above-described further processing of the released fatty acids by lipoxygenases is oxygen-dependent.

[0070] The carboxylester hydrolase used in the method of the present invention is thus preferably a lipase, e.g., a triacylglycerol lipase (EC 3.1.1.1 or EC 3.1.1.3) capable of hydrolyzing lipids into fatty acids and mono- or diglycerides, or a phospholipase (EC 3.1.1.4, EC 3.1.1.32) capable of hydrolyzing diacyl-phospholipids into fatty acids and monoacyl-phospholipids, or a a glycolipase such as galactolipase (EC 3.1.1.26) capable of hydrolyzing diacyl monogalactosides or diacyl digalactosides into the corresponding monoacyl galactosides. In some embodiments, the carboxylester hydrolase may, e.g., be triacylglycerol lipase, EC 3.1.1.3 from Thermomyces lanugi- nosus (formerly Humicola lanuginose) or a carboxyl ester hydrolase with phospholipase A1 (EC 3.1.1.32), galactolipase (EC 3.1.1.26) and triacylglycerol lipase (EC 3.1.1.3.) activity from Fusarium oxysporum.

[0071] An exemplary carboxylester hydrolase suitable for use in the method of the invention may, e.g., comprise an amino acid sequence having SEQ ID NO: 1. The enzyme may also comprise an amino acid sequence having at least 80 %, at least 85 %, at least 90 %, at least 95 %, at least 96 %, at least 97 %, at least 98 %, or at least 99 % sequence identity to SEQ ID NO: 1 , wherein the enzyme is capable of hydrolysing lipids in doughs formed by ground plant products, e.g., as defined herein. The enzyme may also consist of SEQ ID NO: 1.

[0072] Alternatively, the carboxylester hydrolase may also comprise an amino acid sequence having SEQ ID NO: 2. The enzyme may also comprise an amino acid sequence having at least 80 %, at least 85 %, at least 90 %, at least 95 %, at least 96 %, at least 97 %, at least 98 %, or at least 99 % sequence identity to SEQ ID NO: 2, wherein the enzyme is also capable of hydrolyzing lipids in doughs formed by ground plant products, e.g., as defined herein. The enzyme may also consist of SEQ ID NO: 2.

[0073] In a further embodiment, the at least one enzyme may also be another type of hydrolytically active enzyme, such as, e.g., a hemicellulase, i.e. , an enzyme capable of breaking down and hydrolysing hemicellulose. Hemicellulose is a polysaccharide molecule found in the cell walls of plants that is often associated with cellulose but has a distinct composition and structure. Hemicellulose can comprise a variety of sugar building blocks, including xylose, arabinose, mannose and galactose. Examples of hemicellulose include xylan, glucuronoxylan, arabinoxylan, glucomannan, and xyloglucan.

[0074] Preferably, the hemicellulase is endo-1 ,4-p-xylanase, (EC 3.2.1.8), a hemicellulase that degrades the linear polysaccharide xylan into xylose. The hemicellulase may be e.g., the endo-1 ,4-p-xy- lanase of Trichoderma reesei (renamed Hypocrea jecorina'). The hemicellulase used in the method of the present invention may however also be an arabinofuranosidase or a feruloylester- ase.

[0075] The hydrolytically active enzyme may also be a protease, also known as peptidase or proteinase. Proteases are enzymes that catalyse the break-down of proteins into smaller polypeptides or single amino acids, a process known as proteolysis. Proteases may be able to degrade gluten in wheat. Preferably, the protease is derived from a fungus such as Aspergillus oryzae or Aspergillus niger or from Bacillus subtilis or Bacillus licheniformis.

[0076] The at least one hydrolytically active enzyme may also be an amylase. Amylases catalyse the hydrolysis of starch into sugars. Preferably, the amylase is an a-amylase (EC 3.2.1.1). It may, however, also be a p- or glucoamylase (EC 3.2.1.2 or EC 3.2.1.3, respectively).

[0077] The at least one hydrolytically active enzyme added during step a) of the method of the invention may also be a glucanase capable of hydrolysing glycosidic bonds in polysaccharides. Glu- canases, in particular p-glucanases, can degrade the high-molecular-weight p-glucan to low-mo- lecular-weight oligosaccharide fragments, which reduces the viscosity of the p-glucans, promotes cross-linking between gluten molecules, and indirectly improves the gluten network structure in the pasta dough. The at least one hydrolytically active enzyme may also be a cellulase (EC 3.2.1.4) that can catalyse cellulolysis, i.e., the decomposition of cellulose and of some related polysaccharides.

[0078] In further embodiments, the enzyme may also be a transglutaminase (EC 2.3.2.13), e.g., a transglutaminase from Streptomyces mobaraensis, that catalyses the acyl-transfer reaction between e-amino groups of peptide-bound lysine residues and the y-carboxyamide group of peptide- bound glutamine residues. In consequence, individual gluten chains may become permanently cross-linked via iso-peptide bonds (Meerts et al., 2017).

[0079] Therefore, the enzyme of the invention may also comprise an amino acid sequence having SEQ ID NO: 4. Such an enzyme is commercially available as “Sternzym PT 8001” from SternEnzym GmbH & Co. KG. The enzyme may also comprise an amino acid sequence having at least 80 %, at least 85 %, at least 90 %, at least 95 %, at least 96 %, at least 97 %, at least 98 %, or at least 99 % sequence identity to SEQ ID NO: 4, wherein the enzyme is also capable of strengthening the protein matrix in doughs formed by ground plant products, in particular via catalysing the formation of iso-peptide crosslinks, e.g., as defined herein. The enzyme may also consist of SEQ ID NO: 4.

[0080] In the context of the invention, the at least one enzyme that is added in step a) of the method of the invention may also be a combination of at least 2, e.g., at least 3, 4, 5, 6, 7, 8, 9, or at least 10 of the herein disclosed enzymes. Preferably, the at least one enzyme may be a combination of glucose oxidase and a carboxylester hydrolase, preferably a lipase, as defined herein. Such combination of a glucose oxidase and a lipase is commercially available, e.g., as “Pastazym Duo Pure” from Muhlenchemie GmbH & Co. KG. and has been used in some of the experiments described in the Examples below. Based on the results presented herein, it can be assumed that the combined used of an oxidoreductase, such as, e.g., a glucose oxidase, and a carboxylester hydrolase such as, e.g., a lipase may exert at least an additive, preferably a synergistic effect in the method of the invention. Possibly, also other combinations of at least two of the herein disclosed enzymes may exert additive and / or synergistic effects on the quality of the produced pasta. In some embodiments, if the at least one enzyme is a carboxylester hydrolase, e.g. a lipase, or the combination of enzymes comprises such a carboxylester hydrolase, it may be advantageous to further add a lipoxygenase to the pasta dough in step a) of the method according to the invention, which, as described above, can further process the fatty acids released by the carboxylester hydrolases and convert them into peroxides.

[0081] Moreover, the simultaneous use of hydrolytic enzymes like hemicellulases, amylases or proteases may possibly further enhance the structure-forming / cross-linking effects of the carboxylester hydrolases and / or oxidoreductases described herein by increasing the availability of water through the release of pentosan or starch gels as well as hydrated proteins. Typically, the at least one enzyme is added during step a) of the method of the invention at a concentration of about 1-500 ppm (1-500 mg / kg), preferably of about 10-400 ppm (10-400 mg / kg), or of about 50-300 ppm (50-300 mg / kg). However, the optimal enzyme concentration can vary depending on the enzyme used and / or whether more than one enzyme is to be added. For instance, if the at least one enzyme is a glucose oxidase, the enzyme concentration may preferably range between about 100 and 300 ppm (100-300 mg / kg) preferably about 150-200 ppm (150-200 mg / kg). A carboxylester hydrolase as described herein may be mixed with the ingredients during step a) at a preferred concentration of below 100 ppm (100 mg / kg), e.g., at a concentration of about 30-70 ppm (30-70 mg / kg).

[0082] The at least one enzyme is preferably added together with the other ingredients used for preparing the pasta dough prior to mixing. Alternatively, it may be first added either to the water or the ground plant product and premixed, before mixing the ground plant product with water, respectively, and forming the dough.

[0083] In some embodiments, the at least one enzyme is provided, e.g., as a dry powder, such as a lyophilised or spray-dried powder, which is mixed directly with the dry ground plant product before the ground plant product is contacted with the water during step a). This may lead to a particularly homogenous distribution of the enzyme in the prepared pasta dough. The enzyme powder preferably has a particle size of 10-1000 pm, preferably of less than 1000 pm, less than 900 pm, less than 800 pm, less than 700 pm, less than 600 pm or less than less than 500 pm. The particle size should therefore preferably be in the range of 10-500 pm, e.g., of 15-300 pm or 20-200 pm. Most preferably, the at least one enzyme is provided as a powder having an average particle size of 25-150 pm. Optionally, the enzyme may also be provided as a liquid preparation, i.e. it can be dissolved in water and optionally stabilised with, e.g., glycerol, sorbitol and / or salts.

[0084] In addition or alternatively, the at least one enzyme may be dissolved in or added to water prior to being mixed with the ground plant product. Preferably, the enzyme is added to the water that is mixed with the ground plant product for producing the pasta dough. The mixing of the at least one enzyme with the ground plant product should be sufficiently long and thorough to ensure an even distribution of the enzyme. The skilled person will be able to assess when the ground plant product has been sufficiently mixed with the at least one enzyme.

[0085] In further embodiments, the at least one enzyme may also be added after the water and the ground plant product have been mixed to create a dough.

[0086] Another aspect that may significantly affect the activity of the at least one enzyme added during step a) of the method of the invention is the temperature of the water used for producing the pasta dough and thus also the temperature of the pasta dough itself. The optimal choice of the water temperature may depend on various variables, including the temperature of the flour / semolina, the particle size (fine or coarse), the mixing time required to obtain a homogenous pasta dough, which largely depends on the kneading technology in use, the presence of vacuum, the shape and speed of the blades, the types of extruders and dies, the final pasta shape, the type of drying, etc.

[0087] For long pasta with a hollow centre like so-called Bucatini, the water should be rather cold, e.g., at a temperature of about 15-35 °C to prevent the pasta product from flaking where it comes into contact with the support rod. Cold water at temperatures of about 10-25 °C is also preferable for rather fine and tiny, short pasta to prevent the cutting surface from turning white. Hotter water having a temperature of about 40-60 °C, on the other hand, is preferred for the preparation of short pasta with a larger diameter (conchiglie, farfalle, etc.) as well as for long, unpunctured and thin pasta (spaghetti) and for making egg pasta (e.g. tagliatelle).

[0088] However, special care should be taken that the temperature of the final pasta dough does not exceed 45 °C during mixing and kneading in steps a) and b) to prevent the proteins forming the matrix in the pasta dough as well as the at least one added enzyme from denaturing, which would have a negative effect on the pasta quality.

[0089] The optimum reaction temperature in particular of fungal enzymes such as, e.g., the herein disclosed glucose oxidases or lipases is in the range of 30-45 °C. Typically, the temperature of pasta dough during pasta production is about 40 °C. Accordingly, it was to be expected that mixing the ingredients of the pasta dough during step a) at temperatures of around 40 °C would provide optimal reaction conditions for the enzymes used in the method of the invention. Surprisingly however, when the present inventors reduced the temperature during the mixing step a) to merely 20 °C, they observed that a combination of glucose oxidase and lipase still significantly improved the cooking and textural properties of the pasta. As can be seen in the Examples below, use of a combination of glucose oxidase and lipase at merely 20 °C in a method of the present invention resulted in an improvement of the firmness and a reduction of the stickiness of the cooked pasta. Importantly, the enzymes appear to perform at least as efficient at 20 °C as at their optimal reaction temperatures of around 40 °C. By reducing the temperature during the production of the pasta dough, a considerable amount of energy can thus be saved without adversely affecting enzyme activity.

[0090] Accordingly, in a preferred embodiment, step a) and preferably, step b) of the method of the present invention are performed at a temperature below 40 °C, preferably below 35 °C, more preferably below 30 °C, e.g., between 15-30 °C such as, e.g., about 15 °C, 20 °C or 25 °C. In summary, the present invention teaches a novel method for preparing pasta under vacuum conditions that involves the use of at least one enzyme capable of stabilizing the final pasta during cooking. It was surprisingly found that the enzymes disclosed herein, such as glucose oxidase and / or lipase were able to positively influence the stability and stickiness of the prepared and cooked pasta even under full vacuum conditions. This result was completely unexpected, especially with regard to the use of oxidoreductases such as glucose oxidase, which normally relies on oxygen for its catalytic activity.

[0091] Throughout the invention, the term “about” is intended to be understood as ”+ / - 10 %”. If “about” relates to a range, it refers to both lower and upper limit of the range. “A” is intended to mean “one or more”, if not explicitly mentioned otherwise.

[0092] The present invention encompasses, inter alia, the following embodiments:

[0093] 1 . A method for preparing pasta comprising steps of: a) mixing water and a ground plant product to obtain a pasta dough, b) kneading the pasta dough, c) forming the pasta dough into pasta of a desired shape, wherein steps b), c) and, optionally, step a) are performed in a vacuum, wherein, during step a), at least one enzyme selected from the group comprising an oxi- doreductase, a carboxylester hydrolase, a hemicellulase, a glucanase, a cellulase, a transglutaminase, a protease, an amylase and a combination thereof is added to the pasta dough.

[0094] 2. Use of at least one enzyme selected from the group comprising an oxidoreductase, a carboxylester hydrolase, a hemicellulase, a glucanase, a cellulase, a transglutaminase, a protease, an amylase and a combination thereof for preparing pasta in a vacuum.

[0095] 3. Use of at least one enzyme selected from the group comprising an oxidoreductase, a carboxylester hydrolase, a hemicellulase, a glucanase, a cellulase, a transglutaminase, a protease, an amylase and a combination thereof for increasing the firmness and cooking tolerance and / or decreasing the stickiness of cooked pasta that has been prepared in a vacuum.

[0096] 4. The use of any of embodiments 2 or 3, wherein the method of embodiment 1 is used.

[0097] 5. The method of embodiment 1 or the use of any of embodiments 2-4, wherein steps b), c) and, optionally, step a) are performed at a negative gauge pressure of less than -800 hPa. The method of embodiment 1 or the use of any of embodiments 2-4, wherein steps b), c) and, optionally, step a) are performed at a negative gauge pressure of -980 hPa or lower. The method of any of embodiments 1 or 5-6 or the use of any of embodiments 2-6, wherein the at least one enzyme is an oxidoreductase. The method of embodiment 7 or the use of embodiment 7, wherein the oxidoreductase is a glucose oxidase. The method of embodiment 7 or the use of embodiment 7, wherein the oxidoreductase is a sulfhydryl oxidase. The method of embodiment 7 or the use of embodiment 7, wherein the oxidoreductase is a phenol oxidase, preferably a laccase. The method of embodiment 7 or the use of embodiment 7, wherein the oxidoreductase is a peroxidase. The method of embodiment 7 or the use of embodiment 7, wherein the oxidoreductase is a tyrosinase. The method of embodiment 7 or the use of embodiment 7, wherein the oxidoreductase is a lipoxygenase. The method of embodiment 7 or the use of embodiment 7, wherein the oxidoreductase is an amino acid oxidase. The method of any of embodiments 1 or 5-6 or the use of any of embodiments 2-6, wherein the at least one enzyme is a carboxylester hydrolase. The method of embodiment 15 or the use of embodiment 15, wherein the carboxylester hydrolase is a lipase. The method of embodiment 16 or the use of embodiment 16, wherein the lipase is a triacylglycerol lipase. The method of embodiment 16 or the use of embodiment 16, wherein the lipase is a phospholipase. The method of embodiment 16 or the use of embodiment 16, wherein the lipase is a galactolipase. 20. The method of any of embodiments 1 or 5-6 or the use of any of embodiments 2-6, wherein the at least one enzyme is a hemicellulase.

[0098] 21. The method of embodiment 20 or the use of embodiment 20, wherein the hemicellulase is an endo-1 ,4-p-xylanase.

[0099] 22. The method of embodiment 21 or the use of embodiment 21 , wherein the hemicellulase is an arabinofuranosidase.

[0100] 23. The method of embodiment 21 or the use of embodiment 21 , wherein the hemicellulase is a feruloylesterase.

[0101] 24. The method of any of embodiments 1 or 5-6 or the use of any of embodiments 2-6, wherein the at least one enzyme is a protease.

[0102] 25. The method of any of embodiments 1 or 5-6 or the use of any of embodiments 2-6, wherein the at least one enzyme is an amylase.

[0103] 26. The method of any of embodiments 1 or 5-6 or the use of any of embodiments 2-6, wherein the at least one enzyme is a glucanase.

[0104] 27. The method of any of embodiments 1 or 5-6 or the use of any of embodiments 2-6, wherein the at least one enzyme is a cellulase.

[0105] 28. The method of any of embodiments 1 or 5-6 or the use of any of embodiments 2-6, wherein the at least one enzyme is a transglutaminase.

[0106] 29. The method of any of embodiments 1 or 5-28 or the use of any of embodiments 2-28, wherein a combination of at least two enzymes of any of embodiments 7-28 is used.

[0107] 30. The method of any of embodiments 1 or 5-28 or the use of any of embodiments 2-28, wherein a combination of at least three enzymes of any of embodiments 7-28 is used. 31 . The method of any of embodiments 1 or 5-28 or the use of any of embodiments 2-28, wherein a combination of at least four enzymes of any of embodiments 7-28 is used.

[0108] 32. The method of any of embodiments 1 or 5-28 or the use of any of embodiments 2-28, wherein a combination of at least five enzymes of any of embodiments 7-28 is used.

[0109] 33. The method of any of embodiments 1 or 5-28 or the use of any of embodiments 2-28, wherein a combination of at least six enzymes of any of embodiments 7-28 is used. 34. The method of any of embodiments 1 or 5-28 or the use of any of embodiments 2-28, wherein a combination of at least seven enzymes of any of embodiments 7-28 is used.

[0110] 35. The method of any of embodiments 1 or 5-28 or the use of any of embodiments 2-28, wherein a combination of at least eight enzymes of any of embodiments 7-28 is used.

[0111] 36. The method of any of embodiments 1 or 5-28 or the use of any of embodiments 2-28, wherein a combination of at least nine enzymes of any of embodiments 7-28 is used.

[0112] 37. The method of any of embodiments 1 or 5-28 or the use of any of embodiments 2-28, wherein a combination of at least ten enzymes of any of embodiments 7-28 is used.

[0113] 38. The method of any of embodiments 1 or 5-28 or the use of any of embodiments 2-28, wherein the at least one enzyme is a combination of an enzyme of any of embodiments 7-14 and an enzyme of any of embodiments 15-19.

[0114] 39. The method of embodiment 38 or the use of embodiment 38, wherein the at least one enzyme is a combination of a glucose oxidase and a lipase.

[0115] 40. The method of any of embodiments 1 and 5-39 or the use of any of embodiments 2-39, wherein the at least one enzyme is added to the pasta dough at a concentration of 1-500 mg / kg.

[0116] 41. The method of any of embodiments 1 and 5-40 or the use of any of embodiments 2-40, wherein steps a) and b) are performed at a temperature of 15-30 °C.

[0117] 42. The method of any of embodiments 1 and 5-41 or the use of any of embodiments 2-41 , wherein the ground plant product is selected from the group comprising wheat, rye, oat, barley, millet, rice, maize, buckwheat, potatoes, soy, quinoa, lentils, beans, peas, chickpeas, lupins and cassava.

[0118] 43. The method of any of embodiments 1 and 5-42 or the use of any of embodiments 2-42, wherein the ground plant product is ground wheat selected from the group comprising Triticum aestivum, Triticum durum, Triticum dicoccum, Triticum spelta, Triticum monococ- cum and a mixture thereof, wherein the wheat preferably is Triticum aestivum.

[0119] 44. The method of any of embodiments 1 and 5-43 or the use of any of embodiments 2-43, wherein the ground plant product is semolina, flour or a mixture thereof.

[0120] All literature cited herein is herewith fully incorporated. The present invention is further illustrated, but not limited, by the following example. Brief description of the drawings:

[0121] Figure 1 : Schematic overview of an exemplary manufacturing process for pasta in accordance with the method of the invention.

[0122] Figure 2: Determination of the optimal cooking time for the pasta to be fully cooked (according to standard method AACC 66-50.01 of the Cereals & Grains Association) and the calculation of the cooking time for the stress test to analyse the cooking tolerance of the pasta.

[0123] Figure 3: Analysis of the texture parameters bite resistance and stickiness using the T exture Analyser TA XT2 from Stable Micro Systems. (A) Image of the Texture Analyser used for analysing the prepared pasta. 1 : Texture Analyser, 2: PMMA knife, 3: Pasta sample, 4: Sample table (B) The positive curve (‘body’) represents the force required to cut 5 parallel strands of cooked spaghetti in half with the PMMA knife of the Texture Analyser. The negative curve (‘stickiness’) shows the adhesive force when the knife is withdrawn.

[0124] Figure 4: Influence of the vacuum level in combination with Pastazym Duo Pure (glucose oxidase + lipase) on the firmness of cooked pasta prepared from T. aestivum soft wheat flour.

[0125] Figure 5: Effect of glucose oxidase (Pastazym PD1) and glucose oxidase + lipase (Pastazym Duo Pure) on the firmness of spaghetti prepared from T. aestivum soft wheat flour at different cooking times.

[0126] Figure 6: Influence of the vacuum level in combination with Pastazym Duo Pure (glucose oxidase + lipase) on the stickiness of cooked pasta prepared from T. aestivum soft wheat flour.

[0127] Figure 7: Effect of glucose oxidase (Pastazym PD1) and glucose oxidase + lipase (Pastazym Duo Pure) on the stickiness of pasta prepared from T. aestivum soft wheat flour at different cooking times.

[0128] Figure 8: Effect of Pastazym Duo Pure (glucose oxidase + lipase) on the structure of pasta obtained in a production process under full vacuum at -980 mbar

[0129] Figure 9: The influence of temperature on the activity of GOX from Aspergillus niger Figure 10: Influence of the dough temperature during the mixing process in combination with Pastazym Duo Pure (glucose oxidase + lipase) on the firmness of the cooked pasta.

[0130] Figure 11 : Influence of the dough temperature during the mixing process in combination with Pastazym Duo Pure (glucose oxidase + lipase) on the stickiness of the cooked pasta.

[0131] Examples:

[0132] In the following example, the stability and stickiness of cooked pasta produced from T. aestivum flour in a partial or full vacuum pasta production process according to the method of the present invention using the two enzymes glucose oxidase (GOX) and triacylglycerol lipase were tested. The tests were conducted in the research laboratory of Fava S.p.A., one of the world’s largest manufacturers for industrial pasta making systems and plants.

[0133] 1. Production of pasta using the method of the invention

[0134] The pasta was prepared using a test plant of Italian company Fava S.p.A. that allows for the production of pasta under full vacuum conditions. A schematic sequence of the manufacturing process is shown in Figure 1. The T. aestivum wheat flour was first mixed with the enzymes (referred to as Pastazym in Figure 1) in powder form in a mixer. For a test on the pilot plant, 25 kg of wheat flour was used. With a dosage of 140 ppm of Pastazym Duo Pure, 3.5 g of Pastazym Duo Pure were mixed with 25 kg of wheat flour. The mixing time was 5 min. For pasta production, the flour treated with Pastazym Duo Pure was fed into the dosing system leading to the mixer of the Fava pilot plant. The Fava pilot pasta press is a full vacuum press, which means that it is configured in a way that allows mixing (step a of the method of the invention), kneading (step b) and extrusion (step c) to take place in the absence of oxygen. The water temperature and quantity can be regulated. The water temperature of the dosing water for the standard tests was 40 °C. In order to test the influence of the dough temperature, tests with varying dough temperatures (20 °C, 40 °C, 55 °C) were carried out later on. The target moisture content of the dough was 32.5 % in all tests. The mixing time was 10 min. To test the effect of the vacuum level, the vacuum pressure was set to -800 mbar or -980 mbar. After 10 min mixing time, the dough was extruded through a die (1 .89 mm) to produce spaghetti with a diameter of 1.7 mm (dried pasta). The extrusion pressure was 100 bar for all tests. The freshly extruded pasta was hung over the ring system, which is part of the Fava pilot dryer. The spaghetti strands were then cut to length. The pasta was subsequently dried for 180 minutes using the GPL 180 technology (patented by Fava S.p.A.: Bianchi and Fava, 2014, EP 2416669 A1). The maximum drying temperature was 92 °C. After the drying time was reached, the pasta was cooled to room temperature in the cooling system for 20 min.

[0135] For the tests in a partial vacuum, the enzyme-treated flour was first mixed with water (40 °C) in a pre-mixer in the presence of oxygen (i.e. , step a) of the method of the invention was not performed in vacuum). The target dough moisture content was 32.5 %. The mixing time was 9 min. The dough was then transferred directly to the mixer of the Fava full-vacuum pilot plant. The dough was then kneaded further for 2 min at a vacuum pressure of -980 mbar and then extruded.

[0136] 2. Determination of optimal cooking time of pasta produced using the method of the invention

[0137] As a first step, the optimal cooking time of the pasta (spaghetti) produced using the method of the present invention was determined using the standard method AACC 66-50.01 (cf. https: / / www.ce- realsgrains.org / resources / Methods / Pages / 66Semolina_Pasta_NoodleQuality.aspx). To do so, the spaghetti were removed from the cooking water at different time points and placed between two sheets of poly(methyl methacrylate)-glass (PMMA glass). The cooking time required until no visible starchy white stripes were observed when the cooked spaghetti were gently squeezed between the two glass plates was considered to represent the optimal cooking time (Figure 2). Using this assay, the optimal cooking time of the produced pasta was determined to be 11.5 min.

[0138] The overcooking time, i.e., the time until the pasta produced using the method of the invention was considered to be overcooked was calculated by multiplying the optimal cooking time by 1.5 (11.5 min x 1.5 = 17.25 min).

[0139] 3. Cooking tests and texture measurements

[0140] The cooking tests and texture measurements were conducted as follows:

[0141] 1 . 100 g of dried pasta (whole spaghetti) produced as described above were added to 1000 mL of boiling tap water.

[0142] 2. The timer was started when the pasta was completely immersed in the water.

[0143] 3. The pasta was cooked without a lid and was stirred several times during the cooking process.

[0144] 4. Once the optimum cooking time was reached (see above and Figure 2), 200 mL of the cooking water was collected in a beaker to assess the appearance and turbidity of the cooking water. Step 4 was omitted in the experiments testing the overcooking tolerance of the pasta. In this case, step 5 is carried out directly after step 3.

[0145] 5. 500 mL of cold tap water (15 °C) was added to the pot with the pasta and the remaining water to interrupt the cooking process (and thus reduce the fluctuations between the individual measurements). 6. The pasta was immediately afterwards transferred into a sieve to remove the water.

[0146] 7. The pasta was transferred from the sieve on a plate and covered during the measurements.

[0147] 8. After 30 seconds, the texture was measured with the texture analyser. To do so, always five strands of spaghetti were placed directly next to each other on the table of the texture analyser in the centre of the knife (Figure 3).

[0148] 9. Each measurement was repeated three times.

[0149] 10. Before each measurement, the knife of the texture analyser was thoroughly cleaned to remove adhering starch.

[0150] 11. The results are given in the dimension [g] for strength and [g s] for tackiness and fullness.

[0151] 4. Influence of the vacuum level and the addition of glucose oxidase and lipase on the firmness of cooked pasta.

[0152] As can be seen in Figure 4, the use of Pastazym Duo Pure, which contains 70 ppm glucose oxidase (GOX) from Aspergillus nigerand 70 ppm triacylglycerol lipase from Thermomyces lanu- ginosus during pasta production resulted in pasta characterised by increased firmness, provided that the pasta was produced either under partial vacuum or in a full vacuum process at -980 mbar. The firmness is indicative of the cooking stability of the cooked pasta. When using full vacuum at -980 mbar, the effect was particularly noticeable for overcooked pasta suggesting that the use of Pastazym Duo Pure provides elevated cooking resistance. Interestingly, the effects of Pastazym Duo Pure were more pronounced when the pasta was produced under full vacuum at -980 mbar than at -800 mbar.

[0153] As expected, the effect of Pastazym Duo Pure on the cooking stability was higher in a partial vacuum (i.e, when mixing step a) of the method of the invention was performed in the presence of oxygen before the kneading of the dough (step b) under vacuum conditions) compared to a total vacuum (mixing step a) and kneading step b) under vacuum, i.e., -800 or -980 mbar) (Figure 4). However, the positive effect of the enzyme was also evident with full vacuum. In particular, the cooking tolerance at full vacuum was at the level of the cooking tolerance at partial vacuum.

[0154] To better understand whether the effect of Pastazym Duo Pure on the firmness and cooking resistance of the produced pasta was due to the activity of GOX or that of lipase, which are both contained in Pastazym Duo Pure, the firmness of cooked pasta produced under full vacuum at -980 mbar using either again Pastazym Duo Pure (GOX + lipase) or Pastazym PD1 (only GOX) were compared using the method as described above. Use of Pastazym Duo Pure increased the firmness and cooking resistance of the final pasta to a larger extent than Pastazym PD1 , suggesting that this effect was primarily a result of the lipase activity (Figure 5). 5. Influence of the vacuum level in combination with Pastazym Duo Pure on the stickiness of cooked pasta.

[0155] Figure 6 shows that the use of the combination of Pastazym Duo Pure and full vacuum at both -800 mbar and -980 mbar had a significant effect on the stickiness of the cooked pasta (Figure 6): While the vacuum at -980 mbar alone only reduced the stickiness slightly, the combination of the total vacuum with Pastazym Duo Pure resulted in a drastic reduction of the pasta stickiness compared to the stickiness observed when the pasta was prepared under partial vacuum conditions, i.e. in the presence of oxygen during step a) of the method of the invention or in the absence of any enzymes.

[0156] The images of the cooked pasta in Figure 8 underline the results of the texture analysis shown in Figure 6: The best results were achieved when pasta was produced under full vacuum (-980 mbar) using Pastazym Duo Pure.

[0157] To again test whether the effect of Pastazym Duo Pure on the stickiness of the produced pasta was rather due to the activity of GOX or that of lipase during pasta production, the stickiness of cooked pasta produced under full vacuum at -980 mbar using again either Pastazym Duo Pure (GOX + lipase) or Pastazym PD1 (only GOX) was compared using the method as described above. This time, the use of Pastazym PD1 and Pastazym Duo Pure yielded similar results, suggesting that the effect on pasta stickiness was primarily a result of the GOX activity (Figure 7) since otherwise, the additional use of the lipase in Pastazym Duo Pure should have had at least an additive effect.

[0158] 6. Effects of temperature on Pastazym Duo Pure activity.

[0159] The temperature optimum of GOX is between 30-50 °C (Figure 9). A similar temperature optimum has been suggested for other enzymes isolated from fungi, including lipases. However, as can be seen in Figure 11 , reducing the temperature during the mixing of the ingredients for preparing the pasta dough down to 20 °C still led to a reduction of the stickiness of the final pasta after cooking.

[0160] The effects of Pastazym Duo Pure on the firmness and cooking resistance of cooked pasta produced at a low temperature of 20 °C remained unchanged compared to pasta produced at 55 °C or 40 °C (Figure 10). Both results are extremely surprising in view of the fact that the optimum temperature of the enzymes used is normally well above 20 °C.

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Claims

Claims1 . A method for preparing pasta comprising steps of: a) mixing water and a ground plant product to obtain a pasta dough, b) kneading the pasta dough, c) forming the pasta dough into pasta of a desired shape, wherein steps b), c) and, optionally, step a) are performed in a vacuum, wherein, during step a), at least one enzyme selected from the group comprising an oxidoreductase, a carboxylester hydrolase, a hemicellulase, a glucanase, a cellulase, a transglutaminase, a protease, an amylase and a combination thereof is added to the pasta dough.

2. Use of at least one enzyme selected from the group comprising an oxidoreductase, a carboxylester hydrolase, a hemicellulase, a glucanase, a cellulase, a transglutaminase, a protease, an amylase and a combination thereof for preparing pasta in a vacuum.

3. Use of at least one enzyme selected from the group comprising an oxidoreductase, a carboxylester hydrolase, a hemicellulase, a glucanase, a cellulase, a transglutaminase, a protease, an amylase and a combination thereof for increasing the firmness and cooking tolerance and / or decreasing the stickiness of cooked pasta that has been prepared in a vacuum.

4. The use of any of claims 2 or 3, wherein the method of claim 1 is used.

5. The method of claim 1 or the use of any of claims 2-4, wherein steps b), c) and, optionally, step a) are performed at a negative gauge pressure of less than -800 hPa.

6. The method of claim 1 or the use of any of claims claim 2-4, wherein steps b), c) and, optionally, step a) are performed at a negative gauge pressure of -980 hPa or lower.

7. The method of any of claims 1 or 5-6 or the use of any of claims 2-6, wherein the at least one enzyme is an oxidoreductase.

8. The method of claim 7 or the use of claim 7, wherein the oxidoreductase is a glucose oxidase.

9. The method of any of claims 1 or 5-6 or the use of any of claims 2-6, wherein the at least one enzyme is a carboxylester hydrolase, preferably a lipase.

10. The method of any of claims 1 or 5-9 or the use of any of claims 2-9, wherein the at least one enzyme is a combination of a glucose oxidase and a lipase.

11. The method of any of claims 1 and 5-10 or the use of any of claims 2-10, wherein the at least one enzyme is added to the pasta dough at a concentration of 1-500 mg / kg.

12. The method of any of claims 1 and 5-11 or the use of any of claims 4-11 , wherein steps a) and b) are performed at a temperature of 15-30 °C.

13. The method of any of claims 1 and 5-12 or the use of any of claims 4-12, wherein the ground plant product is selected from the group comprising wheat, rye, oat, barley, millet, rice, maize, buckwheat, potatoes, soy, quinoa, lentils, beans, peas, chickpeas, lupins and cassava.

14. The method of any of claims 1 and 5-13 or the use of any of claims 4-13, wherein the ground plant product is ground wheat selected from the group comprising Triticum aestivum, Triticum durum, Triticum dicoccum, Triticum spelta, Triticum monococcum and a mixture thereof, wherein the wheat preferably is Triticum aestivum.

15. The method of any of claims 1 and 5-14 or the use of any of claims 4-14, wherein the ground plant product is semolina, flour or a mixture thereof.

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