EP0513709B2 - Enzymatic method for reducing the amount of phosphorous-containing components in vegetable and animal oils - Google Patents
Enzymatic method for reducing the amount of phosphorous-containing components in vegetable and animal oils Download PDFInfo
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- EP0513709B2 EP0513709B2 EP92107888A EP92107888A EP0513709B2 EP 0513709 B2 EP0513709 B2 EP 0513709B2 EP 92107888 A EP92107888 A EP 92107888A EP 92107888 A EP92107888 A EP 92107888A EP 0513709 B2 EP0513709 B2 EP 0513709B2
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- oil
- ppm
- phospholipase
- vegetable
- phosphorus
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- 239000008158 vegetable oil Substances 0.000 title claims description 9
- 239000010775 animal oil Substances 0.000 title claims description 8
- 235000013311 vegetables Nutrition 0.000 title description 4
- 238000006911 enzymatic reaction Methods 0.000 title description 3
- BHEPBYXIRTUNPN-UHFFFAOYSA-N hydridophosphorus(.) (triplet) Chemical compound [PH] BHEPBYXIRTUNPN-UHFFFAOYSA-N 0.000 title 1
- 239000003921 oil Substances 0.000 claims description 45
- 235000019198 oils Nutrition 0.000 claims description 44
- 102000004190 Enzymes Human genes 0.000 claims description 34
- 108090000790 Enzymes Proteins 0.000 claims description 34
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 claims description 31
- 239000011574 phosphorus Substances 0.000 claims description 31
- 229910052698 phosphorus Inorganic materials 0.000 claims description 31
- 102000015439 Phospholipases Human genes 0.000 claims description 26
- 108010064785 Phospholipases Proteins 0.000 claims description 26
- ZIIUUSVHCHPIQD-UHFFFAOYSA-N 2,4,6-trimethyl-N-[3-(trifluoromethyl)phenyl]benzenesulfonamide Chemical compound CC1=CC(C)=CC(C)=C1S(=O)(=O)NC1=CC=CC(C(F)(F)F)=C1 ZIIUUSVHCHPIQD-UHFFFAOYSA-N 0.000 claims description 24
- 238000000034 method Methods 0.000 claims description 17
- 239000000243 solution Substances 0.000 claims description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 14
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 12
- 241001465754 Metazoa Species 0.000 claims description 9
- 239000000203 mixture Substances 0.000 claims description 8
- 238000003756 stirring Methods 0.000 claims description 8
- 235000015112 vegetable and seed oil Nutrition 0.000 claims description 8
- 239000002253 acid Substances 0.000 claims description 7
- 239000007864 aqueous solution Substances 0.000 claims description 4
- 238000006243 chemical reaction Methods 0.000 claims description 4
- 230000007515 enzymatic degradation Effects 0.000 claims description 4
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 claims description 3
- 230000015572 biosynthetic process Effects 0.000 claims description 3
- 239000011575 calcium Substances 0.000 claims description 3
- 229910052791 calcium Inorganic materials 0.000 claims description 3
- 239000000839 emulsion Substances 0.000 claims description 3
- 239000004615 ingredient Substances 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 230000035484 reaction time Effects 0.000 claims description 3
- 239000003995 emulsifying agent Substances 0.000 claims description 2
- 238000000926 separation method Methods 0.000 claims description 2
- BVKZGUZCCUSVTD-UHFFFAOYSA-N carbonic acid Chemical compound OC(O)=O BVKZGUZCCUSVTD-UHFFFAOYSA-N 0.000 claims 2
- 229920000715 Mucilage Polymers 0.000 claims 1
- 239000000853 adhesive Substances 0.000 claims 1
- 229910052783 alkali metal Inorganic materials 0.000 claims 1
- 150000001340 alkali metals Chemical class 0.000 claims 1
- 239000011369 resultant mixture Substances 0.000 claims 1
- YWYZEGXAUVWDED-UHFFFAOYSA-N triammonium citrate Chemical compound [NH4+].[NH4+].[NH4+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O YWYZEGXAUVWDED-UHFFFAOYSA-N 0.000 claims 1
- 229940088598 enzyme Drugs 0.000 description 33
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 10
- 230000000694 effects Effects 0.000 description 10
- 150000003904 phospholipids Chemical class 0.000 description 8
- 239000012071 phase Substances 0.000 description 6
- 239000007857 degradation product Substances 0.000 description 5
- 229910052742 iron Inorganic materials 0.000 description 5
- 239000000787 lecithin Substances 0.000 description 5
- 235000010445 lecithin Nutrition 0.000 description 5
- 239000003549 soybean oil Substances 0.000 description 5
- 235000012424 soybean oil Nutrition 0.000 description 5
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 description 4
- 241000196324 Embryophyta Species 0.000 description 4
- 108091005804 Peptidases Proteins 0.000 description 4
- 102000035195 Peptidases Human genes 0.000 description 4
- 102000000571 Phospholipases A Human genes 0.000 description 4
- 108010002176 Phospholipases A Proteins 0.000 description 4
- 239000008157 edible vegetable oil Substances 0.000 description 4
- 229940067606 lecithin Drugs 0.000 description 4
- ASWBNKHCZGQVJV-UHFFFAOYSA-N (3-hexadecanoyloxy-2-hydroxypropyl) 2-(trimethylazaniumyl)ethyl phosphate Chemical compound CCCCCCCCCCCCCCCC(=O)OCC(O)COP([O-])(=O)OCC[N+](C)(C)C ASWBNKHCZGQVJV-UHFFFAOYSA-N 0.000 description 3
- 108010065511 Amylases Proteins 0.000 description 3
- 102000013142 Amylases Human genes 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 235000019418 amylase Nutrition 0.000 description 3
- 235000014113 dietary fatty acids Nutrition 0.000 description 3
- 239000000194 fatty acid Substances 0.000 description 3
- 229930195729 fatty acid Natural products 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000007670 refining Methods 0.000 description 3
- 239000004094 surface-active agent Substances 0.000 description 3
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 2
- 102000004882 Lipase Human genes 0.000 description 2
- 108090001060 Lipase Proteins 0.000 description 2
- 239000004367 Lipase Substances 0.000 description 2
- 108010019160 Pancreatin Proteins 0.000 description 2
- 235000019484 Rapeseed oil Nutrition 0.000 description 2
- 102000014384 Type C Phospholipases Human genes 0.000 description 2
- 108010079194 Type C Phospholipases Proteins 0.000 description 2
- 239000000654 additive Substances 0.000 description 2
- 229940025131 amylases Drugs 0.000 description 2
- 239000008346 aqueous phase Substances 0.000 description 2
- 239000001110 calcium chloride Substances 0.000 description 2
- 229910001628 calcium chloride Inorganic materials 0.000 description 2
- 150000001732 carboxylic acid derivatives Chemical class 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 239000004927 clay Substances 0.000 description 2
- 239000008162 cooking oil Substances 0.000 description 2
- 239000010779 crude oil Substances 0.000 description 2
- 235000019421 lipase Nutrition 0.000 description 2
- 210000004185 liver Anatomy 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 210000000496 pancreas Anatomy 0.000 description 2
- 229940055695 pancreatin Drugs 0.000 description 2
- 235000019833 protease Nutrition 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 239000007858 starting material Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000004382 Amylase Substances 0.000 description 1
- 101100352919 Caenorhabditis elegans ppm-2 gene Proteins 0.000 description 1
- 102000005575 Cellulases Human genes 0.000 description 1
- 108010084185 Cellulases Proteins 0.000 description 1
- 241000271532 Crotalus Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 102000011720 Lysophospholipase Human genes 0.000 description 1
- 108020002496 Lysophospholipase Proteins 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- 241000228150 Penicillium chrysogenum Species 0.000 description 1
- 102000011420 Phospholipase D Human genes 0.000 description 1
- 108090000553 Phospholipase D Proteins 0.000 description 1
- 102000045595 Phosphoprotein Phosphatases Human genes 0.000 description 1
- 108700019535 Phosphoprotein Phosphatases Proteins 0.000 description 1
- 102000004160 Phosphoric Monoester Hydrolases Human genes 0.000 description 1
- 108090000608 Phosphoric Monoester Hydrolases Proteins 0.000 description 1
- 108010059820 Polygalacturonase Proteins 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 240000005384 Rhizopus oryzae Species 0.000 description 1
- 235000013752 Rhizopus oryzae Nutrition 0.000 description 1
- 241000270295 Serpentes Species 0.000 description 1
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 1
- 235000019486 Sunflower oil Nutrition 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 239000008351 acetate buffer Substances 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 238000013019 agitation Methods 0.000 description 1
- 150000001447 alkali salts Chemical class 0.000 description 1
- 150000003863 ammonium salts Chemical class 0.000 description 1
- 230000003625 amylolytic effect Effects 0.000 description 1
- 239000003659 bee venom Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- FNAQSUUGMSOBHW-UHFFFAOYSA-H calcium citrate Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O.[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O FNAQSUUGMSOBHW-UHFFFAOYSA-H 0.000 description 1
- 239000001354 calcium citrate Substances 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 238000005352 clarification Methods 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 238000010924 continuous production Methods 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 238000004945 emulsification Methods 0.000 description 1
- 230000001804 emulsifying effect Effects 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 108010093305 exopolygalacturonase Proteins 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000003925 fat Substances 0.000 description 1
- 235000019197 fats Nutrition 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 210000004907 gland Anatomy 0.000 description 1
- 125000005456 glyceride group Chemical group 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- 159000000003 magnesium salts Chemical class 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 230000002351 pectolytic effect Effects 0.000 description 1
- 150000002978 peroxides Chemical class 0.000 description 1
- 230000005501 phase interface Effects 0.000 description 1
- 150000008103 phosphatidic acids Chemical class 0.000 description 1
- 235000020777 polyunsaturated fatty acids Nutrition 0.000 description 1
- 229940024999 proteolytic enzymes for treatment of wounds and ulcers Drugs 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 238000007127 saponification reaction Methods 0.000 description 1
- 239000002795 scorpion venom Substances 0.000 description 1
- 239000003998 snake venom Substances 0.000 description 1
- 239000000344 soap Substances 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- -1 sodium citrate) Chemical class 0.000 description 1
- 239000002600 sunflower oil Substances 0.000 description 1
- 235000013337 tricalcium citrate Nutrition 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
- 235000019871 vegetable fat Nutrition 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B3/00—Refining fats or fatty oils
- C11B3/003—Refining fats or fatty oils by enzymes or microorganisms, living or dead
Definitions
- the invention relates to a method for reducing the content of phosphorus-containing components in edible oils by enzymatic degradation and separation of the degradation products.
- edible oils includes vegetable and animal, preferably pre-degummed oils.
- the NHP are created by the action of plant enzymes. These are used in the "Alcon process” inactivated by steam treatment of the soy flakes, so that the formation of the NHP is prevented and at the wet degumming of the crude oil the phosphatide content can be almost completely removed.
- NHP NHP-degummed oil
- aqueous surfactant solutions are extracted, however, one does not usually get below 30 ppm phosphorus. It is more successful Treatment with acids or alkalis, however, requires many steps.
- Example 1 is a degummed oil at a temperature of 60 ° C for 48 hours treated with an enzyme solution with 3U phospholipase A activity.
- the phosphorus content is reduced to 1300 ppm phospholipid. Only after treatment at 105 ° C and 30 Torr with the addition of clay, the phosphorus content can be reduced to 40 ppm phospholipid.
- Example 3 degummed oil with a phosphorus content of 1638 ppm phospholipid 30 minutes, 1 hour, 2 hours or 4 hours at 50 ° C with a high enzyme concentration and with the addition of 100%, 500%, 1000% or 2000% water, based on the amount of oil treated.
- the method for treating oil known from JP-A-2-153997 has the Disadvantage that it requires a very long reaction time (48 hours) and the phosphorus content is not sufficiently reduced becomes.
- the variant in which a further reduction in the phosphorus content possible, in which clay is added to the reaction mixture, also has the disadvantage that the process must be carried out in two stages that the second stage takes place at a relatively high temperature and that a vacuum is applied must become. An additional filtration stage is also required.
- the heating of the oil to a temperature above 80 ° C is disadvantageous. Oxidation reactions take place in the oil and it discolors what to use as an cooking oil is extremely disadvantageous.
- the nutritional value of oils partially with a high content of polyunsaturated fatty acids when heated lost.
- Example 3 The process described in Example 3 has the disadvantage that large enzyme and amounts of water are required, which are determined by the Oil must be separated again. Such conditions are technical and not economically sensible, since huge amounts of waste are generated and there is a risk that stable oil-in-water emulsions form. In addition, with a Except the phosphorus level is not sufficiently lowered, even at a very high one Phospholipase A activity, except for one example where 500% water, based on the oil is used.
- JP-A 2-49593 describes a similar enzyme treatment of oils, which, however, does not the degumming of the oil, but aimed at the extraction of lysolecithin.
- the process according to EP-A 328 789 is also about the conversion of soybean oil lecithin to lysolecithin by phospholipase A in the production of mayonnaise-like Products.
- the aim of the invention is to further reduce the phosphorus and iron content Ingredients in pre-degummed animal or vegetable oils to a phosphorus content below 5 ppm due to enzymatic degradation.
- the drastic reduction in the phosphorus content is surprising in that the phospholipases A 1 , A 2 and B have a pH optimum in the neutral to weakly alkaline range.
- the process goal cannot be achieved with phospholipase C or D.
- the invention therefore relates to a process for reducing the content of phosphorus-containing constituents in animal or vegetable oils by enzymatic degradation by means of a phospholipase.
- a pre-degummed animal or vegetable oil with a phosphorus content of 50 to 250 ppm is stirred with an organic carboxylic acid and the pH of the mixture formed is adjusted to 4 to 6; an enzyme solution containing phospholipase A 1 , A 2 or B is added to the mixture in a mixing vessel with turbulent stirring and formation of fine droplets, an emulsion containing 0.5 to 5% by weight of water, based on the oil, is formed, which is passed through at least one subsequent reaction vessel with turbulent agitation for a reaction time of 0.1 to 10 hours at temperatures in the range from 20 to 80 ° C., and the treated oil, after separating off the aqueous solution, has a phosphorus content below 5 ppm having.
- the starting material is therefore preferably pre-degummed oils, especially edible oils, which are generally characterized by a phosphorus content between 50 and 250 ppm. Oils with fluctuating quality can be processed on the same system. Pre-degummed oils, especially sunflower oil; Rapeseed oil and especially soybean oil are used. It is not necessary to dry the oil beforehand.
- the phospholipase is advantageously used in an aqueous solution which is as fine as possible in the oil is emulsified.
- the enzymatic reaction is expected to occur at the interface between the oil phase and the Water phase take place. It is mixed intensively, e.g. through turbulent stirring and additionally promoted by the addition of surfactants.
- the degradation products of the NHP have a higher hydrophilicity and therefore go into the water phase. They are therefore, like the metal ions, simultaneously with the Water phase removed from the oil.
- Phospholipase A 1 , A 2 and B are known enzymes; (See Pardun, loc.cit. Seiton 135-141). Phospholipase A 1 cleaves the fatty acid ester group on the G 1 atom of a phospholipid molecule. It is found, for example, in rat liver and in the pancreas. An enzyme with phospholipase A 1 activity could be isolated from mold cultures of Rhizopus arrhizus.
- Phospholipase A 2 formerly also called lecithinase A, cleaves the fatty acid ester group on the C 2 atom of a phospholipid molecule. It occurs, mostly in association with other phospholipases, in almost all animal and plant cells. It is abundant in the snake venom of the rattlesnake and the cobra, as well as in the bee and scorpion venom. Technically, it can be obtained from pancreatic glands after activity-inhibiting accompanying proteins have been broken down with trypsin.
- Phospholipase B is widespread in nature. It acts on the lysolecithin caused by the action of phospholipase A 1 by splitting off the second fatty acid ester residue. In some cases it is also regarded as a mixture of the phospholipases A 1 and A 2 . It occurs in rat liver and is also produced by some molds, such as Penicillium notatum.
- Phospholipase A 2 and B are available as commercial products.
- the use of purified enzymes is generally not necessary for technical applications.
- a phospholipase preparation which is obtained from ground pancreatic pulp and which contains above all phospholipase A 2 is suitable for the process of the invention.
- the enzyme is used in amounts of 0.001 to 1% by weight, based on oil.
- a good distribution of the enzyme in the oil is ensured if it is dissolved in an amount of water of 0.5 to 5% by weight, based on oil, and emulsified in this form in the oil into droplets of less than 10 ⁇ m in diameter (weight average) becomes.
- Turbulent stirring at radial speeds above 100 cm / s has proven itself. Instead, the oil can be circulated in the reactor using an external centrifugal pump. The enzymatic reaction can also be promoted by means of ultrasound.
- the enzyme effect is achieved by adding an acid, preferably an organic carboxylic acid, increased, which is added before the enzyme treatment, Citric acid is preferred. It can be in the form of the free acid or in combination as a buffer system with their salt, such as an alkali salt (e.g. sodium citrate), an alkaline earth salt (e.g. calcium citrate) or a Ammonium salt can be used. Suitable amounts are 0.01-1% by weight, based on on oil, optimally 0.1% by weight.
- the pH is adjusted to a value of 3 to 7, preferably 4 to 6, with the acid. The The optimum is around pH 5: Surprisingly, this pH is also optimal when the Phospholipase is used in the form of an enzyme complex from the pancreas.
- the pancreatic enzyme complex otherwise has a pH optimum of 8 and is hardly effective at pH 5. Apparently it turns out to be Phase interface where the enzyme action occurs, a higher pH than within the aqueous phase a.
- emulsifying additives are helpful.
- Water-soluble emulsifiers in particular those with an HLB value above 9, such as sodium dodecyl sulfate, are suitable. It is effective in an amount of, for example, 0.001% by weight based on oil if it is added to the enzyme solution prior to emulsification in the oil.
- Protein additives can be advantageous due to a certain surfactant effect.
- the temperature in the enzyme treatment is not critical. Temperatures are between 20 and 80 ° C suitable. A temperature of 50 ° C is optimal, but it can also be briefly heated to 70 ° C. The duration of treatment depends on the temperature and can be shorter with increasing temperature being held. Times of 0.1 to 10, preferably 1 to 5 hours are usually sufficient.
- a step program in which the first step at a temperature of 40 to 60 ° C and the second stage is carried out at a higher temperature in the range of 50 to 80 ° C. For example, the mixture is first stirred at 50 ° C. for 3 hours, then at 75 ° C. for one hour.
- the enzyme solution together with the degradation products contained therein the NHP is separated from the oil phase, preferably by centrifugation. Because the enzymes get through a high stability and the amount of degradation products absorbed is low same enzyme solution can be reused several times.
- the method is preferably carried out continuously.
- the oil is emulsified in a first mixing vessel with the enzyme solution, in one or several subsequent reaction vessels, if necessary with increasing temperature, under turbulent Movement allowed to react, and then the aqueous enzyme solution separated in a centrifuge.
- a part of it can be running replaced with fresh enzyme solution and the rest returned to the process.
- the oil obtained has a phosphorus content below 5 ppm and is therefore for physical refining suitable for cooking oil. Thanks to the low iron content that has been achieved, it has good preconditions for Refining to achieve high resistance to oxidation.
- 1 l wet degummed soybean oil with a residual phosphorus content of 110 ppm is heated to 75 ° C in a round bottom flask. With vigorous stirring with a 5 cm diameter paddle stirrer at 700 rpm, 10 ml of water containing 1 g of citric acid are added and stirring is continued for 1 hour. The mixture is then cooled to 40 ° C. and a solution of 0.1 g of phospholipase A 2 of the quality given in Example 1 and 50 mg of calcium chloride in 20 ml of 0.1 molar acetate buffer pH 5.5 are added. After a further 5 hours of intensive stirring, the water phase is centrifuged off. The oil obtained contains 2 ppm P and is suitable for physical refining.
- Example 2 The procedure according to Example 1 is repeated with the difference that 1 g of a pancreatic preparation (pancreatin, 800 phospholipase units / g) is used instead of phospholipase A 2 .
- the preparation contains phospholipase A 2 , proteinase, amylase, lipase.
- the phosphorus content drops below 1 ppm. Due to the influence of lipase, there is only a slight increase in the acid number from 0.91 to 1.49.
- 9 l wet-degummed rapeseed oil with a phosphorus content of 72 ppm is mixed with a solution of 8.6 g citric acid in 250 ml water and heated to 60 ° C.
- the mixture is homogenized by circulating it once per minute using an external centrifugal pump.
- the pH of the aqueous phase is adjusted to 5.0 with 30 g of 10% sodium hydroxide solution.
- 9 g of phospholipase A 2 with an activity of 400 U / g are added together with a little calcium chloride and the mixture is circulated for 3 hours at 60 ° C. in the manner indicated. After centrifugation, a phosphorus content of 3 ppm is found.
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- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Microbiology (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biochemistry (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Edible Oils And Fats (AREA)
- Fats And Perfumes (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Enzymes And Modification Thereof (AREA)
Description
Die Erfindung betrifft ein Verfahren zur Verminderung des Gehaltes an phosphorhaltigen Bestandteilen in Speiseölen durch enzymatischen Abbau und Abtrennung der Abbauprodukte. Der Begriff Speiseöle umfaßt pflanzliche und tierische, vorzugsweise vorentschleimte Öle.The invention relates to a method for reducing the content of phosphorus-containing components in edible oils by enzymatic degradation and separation of the degradation products. The term edible oils includes vegetable and animal, preferably pre-degummed oils.
Rohes Sojaöl und andere pflanzliche Rohöle werden einer Vorentschleimung unterworfen, bei der Phosphatide, wie Lecithin, und andere hydrophile Nebenbestandteile entfernt werden. Geschieht dies durch Extrahieren mit Wasser, spricht man auch von Naßentschleimung. Bei dieser Behandlung verbleibt ein Teil der Phosphatide im Öl, der unter dem Begriff "Nicht-hydratisierbare Phosphatide" (NHP) zusammengefaßt wird. Für die Herstellung von Speiseölen ist es unerläßlich, diesen Anteil zu entfernen; nach herrschender Meinung soll der Phosphorgehalt 5 ppm nicht überschreiten. (Vgl. Hermann Pardun, "Die Pflanzenlecithine", Verlag für chemische Industrie H. Ziolkowsky KG, Augsburg. 1988, Seiten 181-194)Crude soybean oil and other vegetable crude oils are subjected to a pre-degum treatment, in which Phosphatides such as lecithin and other hydrophilic minor components are removed. This happens through Extracting with water is also called wet degumming. Part of this treatment remains the phosphatide in oil, which is summarized under the term "non-hydratable phosphatide" (NHP) becomes. For the production of edible oils, it is essential to remove this portion; after ruling Opinion, the phosphorus content should not exceed 5 ppm. (See Hermann Pardun, "The Plant Lecithins", Publishing house for chemical industry H. Ziolkowsky KG, Augsburg. 1988, pages 181-194)
Die NHP entstehen durch die Wirkung pflanzeneigener Enzyme. Diese werden beim "Alcon-Verfahren" durch Dampfbehandlung der Sojaflocken inaktiviert, so daß die Bildung der NHP unterbunden wird und bei der Naßentschleimung des rohen Öles der Phosphatidanteil nahezu vollständig entfernt werden kann.The NHP are created by the action of plant enzymes. These are used in the "Alcon process" inactivated by steam treatment of the soy flakes, so that the formation of the NHP is prevented and at the wet degumming of the crude oil the phosphatide content can be almost completely removed.
Aus dem vorentschleimten Öl kann mittels wäßriger Tensidlösungen ein wesentlicher Teil der NHP extrahiert werden, jedoch kommt man in der Regel nicht unter 30 ppm Phosphor. Erfolgreicher ist die Behandlung mit Säuren oder Alkalien, erfordert jedoch viele Arbeitsschritte.A substantial part of the NHP can be removed from the pre-degummed oil using aqueous surfactant solutions are extracted, however, one does not usually get below 30 ppm phosphorus. It is more successful Treatment with acids or alkalis, however, requires many steps.
Bekannt ist die Behandlung pflanzlicher und tierischer Öle mit Enzymen, wodurch enzymatisch spaltbare Bestandteile zu wasserlöslichen, leicht extrahierbaren Stoffen abgebaut werden sollen. So werden nach DE-A 16 17 001 Fette für die Seifenherstellung mit proteolytischen Enzymen desodoriert. Zur Klärung von Pflanzenölen werden gemäß GB 1 440 462 amylolytische und pektolytische Enzyme eingesetzt. Nach EP-A 70 269 werden tierische oder pflanzliche Fette oder Öle im rohen, halbaufbereiteten oder raffinierten Zustand mit einem oder mehreren Enzymen behandelt, um alle Bestandteile, die keine Glyceride sind, zu spalten und abzutrennen. Als geeignete Enzyme werden Phosphatasen, Pektinasen, Zellulasen, Amylasen und Proteasen erwähnt. Als Beispiel einer Phosphatase wird Phospholipase C genannt. Die Anwendung von Enzymen zur Vollentlecithinierung oder Totalentschleimung, wie man die NHP-Entfernung aus vorentschleimten Ölen auch nennt, ist nicht bekannt.It is known to treat vegetable and animal oils with enzymes, which makes them enzymatic fissile components are to be broken down into water-soluble, easily extractable substances. So be according to DE-A 16 17 001 deodorized fats for soap production with proteolytic enzymes. For clarification Vegetable oils are used according to GB 1 440 462 amylolytic and pectolytic enzymes. After EP-A 70 269 describes animal or vegetable fats or oils in raw, semi-prepared or refined Condition treated with one or more enzymes to remove all ingredients that are not glycerides split and separate. Suitable enzymes are phosphatases, pectinases, cellulases, amylases and proteases mentioned. Phospholipase C is mentioned as an example of a phosphatase. The application of Enzymes for full deglecination or total degumming, how to degum the NHP removal Oil is also not known.
Die Natur der NHP ist nicht genau bekannt. Nach Pardun (loc.cit) handelt es sich um Lysophosphatide und Phosphatidsäuren bzw. daraus gebildete Calcium- und Magnesium-Salze, die durch den Abbau von Phosphatiden unter der Einwirkung pflanzeneigener Phospholipasen entstehen.The nature of the NHP is not exactly known. According to Pardun (loc.cit) these are lysophosphatides and phosphatidic acids or calcium and magnesium salts formed therefrom, which are produced by the degradation of Phosphatides arise under the influence of the plant's own phospholipases.
Aus der JP-A 2-153997 ist es bekannt, rohes oder vorentschleimtes Öl mit einem Enzym, das Phospholipase-A-Aktivität aufweist, zu behandeln. Eine Regulierung des pH-Wertes ist nicht vorgesehen.From JP-A 2-153997 it is known to use crude or degummed oil with an enzyme that Has phospholipase A activity to treat. Regulation of the pH is not provided.
Hinsichtlich des Phorphorgehaltes des Ausgangsmaterials als auch des Phosphorgehaltes des Endproduktes finden sich in der Beschreibung keine Angaben. Gemäß Beispiel 1 wird ein entschleimtes Öl bei einer Temperatur von 60°C 48 Stunden mit einer Enzymlösung mit 3U Phospholipase-A-Aktivität behandelt. Der Phorphorgehalt wird dabei auf 1300 ppm Phospholipid verringert. Erst durch eine Behandlung bei 105°C und 30 Torr unter Zugabe von Ton kann der Phosphorgehalt auf 40 ppm Phospholipid verringert werden. Gemäß Beispiel 3 wird entschleimtes Öl mit einem Phosphorgehalt von 1638 ppm Phospholipid 30 Minuten, 1 Stunde, 2 Stunden oder 4 Stunden bei 50°C mit einer hohen Enzymkonzentration und unter Zugabe von 100%, 500%, 1000% oder 2000% Wasser, bezogen auf die Ölmenge, behandelt. Dabei wird der Phosphorgehalt nach 4 Stunden je nach den Bedingungen auf 266 ppm, 81 ppm, 153 ppm Phospholipid, 206 ppm Phospholipid oder 179 ppm Phospholipid verringert.With regard to the phosphorus content of the starting material as well as the phosphorus content There is no information in the description of the end product. According to Example 1 is a degummed oil at a temperature of 60 ° C for 48 hours treated with an enzyme solution with 3U phospholipase A activity. The phosphorus content is reduced to 1300 ppm phospholipid. Only after treatment at 105 ° C and 30 Torr with the addition of clay, the phosphorus content can be reduced to 40 ppm phospholipid. According to Example 3, degummed oil with a phosphorus content of 1638 ppm phospholipid 30 minutes, 1 hour, 2 hours or 4 hours at 50 ° C with a high enzyme concentration and with the addition of 100%, 500%, 1000% or 2000% water, based on the amount of oil treated. The phosphorus content after 4 hours depending on the conditions to 266 ppm, 81 ppm, 153 ppm phospholipid, 206 ppm phospholipid or 179 ppm phospholipid decreased.
Das aus der JP-A-2-153997 bekannte Verfahren zur Behandlung von Öl besitzt den Nachteil, daß es eine sehr lange Reaktionszeit (48 Stunden) erfordert, und der Phosphorgehalt nicht ausreichend verringert wird. Die Variante, bei der eine weitere Verringerung des Phosphorgehaltes möglich wird, bei der Ton zu dem Reaktionsgemisch zugegeben wird, besitzt zusätzlich den Nachteil, daß das Verfahren in zwei Stufen durchgeführt werden muß, daß die zweite Stufe bei relativ hoher Temperatur abläuft und daß ein Vakuum angelegt werden muß. Außerdem ist eine zusätzliche Filtrationsstufe erforderlich. Das Erhitzen des Öls auf eine Temperatur über 80°C ist nachteilig. In dem Öl laufen Oxidationsreaktionen ab, und es verfärbt sich, was für die Verwendung als Speiseöl äußerst nachteilig ist. Außerdem geht der ernährungsphysiologische Wert von Ölen mit hohem Gehalt an mehrfach ungesättigten Fettsäuren beim Erhitzen teilweise verloren.The method for treating oil known from JP-A-2-153997 has the Disadvantage that it requires a very long reaction time (48 hours) and the phosphorus content is not sufficiently reduced becomes. The variant in which a further reduction in the phosphorus content possible, in which clay is added to the reaction mixture, also has the disadvantage that the process must be carried out in two stages that the second stage takes place at a relatively high temperature and that a vacuum is applied must become. An additional filtration stage is also required. The heating of the oil to a temperature above 80 ° C is disadvantageous. Oxidation reactions take place in the oil and it discolors what to use as an cooking oil is extremely disadvantageous. In addition, the nutritional value of oils partially with a high content of polyunsaturated fatty acids when heated lost.
Das in Beispiel 3 beschriebene Verfahren besitzt den Nachteil, daß große Enzym- und Wassermengen erforderlich sind, die nach Beedingung des Verfahrens von dem Öl wieder abgetrennt werden müssen. Solche Bedingungen sind technisch und wirtschaftlich nicht sinnvoll, da riesige Abfallmengen entstehen und die Gefahr besteht, daß sich stabile Öl-in-Wasser-Emulsionen bilden. Außerdem wird mit einer Ausnahme der Phosphorgehalt nicht ausreichend erniedrigt, auch bei einer sehr hohen Phospholipase-A-Aktivität, mit Ausnahme eines Beispiels, bei dem 500% Wasser, bezogen auf das Öl, eingesetzt wird.The process described in Example 3 has the disadvantage that large enzyme and amounts of water are required, which are determined by the Oil must be separated again. Such conditions are technical and not economically sensible, since huge amounts of waste are generated and there is a risk that stable oil-in-water emulsions form. In addition, with a Except the phosphorus level is not sufficiently lowered, even at a very high one Phospholipase A activity, except for one example where 500% water, based on the oil is used.
In der JP-A 2-49593 wird eine ähnliche Enzymbehandlung von Ölen beschrieben, die indessen nicht auf die Entschleimung des Öls, sondern auf die Gewinnung von Lysolecithin abzielt. Dabei ist die Einstellung besonderer pH-Werte überflüssig. Auch beim Verfahren gemäß EP-A 328 789 geht es um die Umwandlung des Lecithins des Sojaöls zu Lysolecithin durch Phospholipase A bei der Herstellung von mayonnaiseartigen Produkten.JP-A 2-49593 describes a similar enzyme treatment of oils, which, however, does not the degumming of the oil, but aimed at the extraction of lysolecithin. Here is the setting special pH values are superfluous. The process according to EP-A 328 789 is also about the conversion of soybean oil lecithin to lysolecithin by phospholipase A in the production of mayonnaise-like Products.
Ziel der Erfindung ist die weitere Verminderung des des Gehaltes an phosphor- und eisenhaltigen Bestandteilen in vorentschleimten tierischen oder pflanzlichen Ölen auf einen Phosphorgehalt unter 5 ppm durch enzymatischen Abbau.The aim of the invention is to further reduce the phosphorus and iron content Ingredients in pre-degummed animal or vegetable oils to a phosphorus content below 5 ppm due to enzymatic degradation.
Es wurde gefunden, daß die Behandlung von tierischen oder pflanzlichen Ölen, die einen Phosphorgehalt von 50 bis 250 ppm haben, mit einer Phospholipase A1, A2 oder B in einem kontinuierlichen Verfahren zu Phosphorgehalten unter 5 ppm führt, wenn das Enzym in Form einer zu feinen Tröpfchen emulgierten wäßrigen Lösung bei einem pH-Wert von 4 bis 6 behandelt wird. Es wurden Phorphorgehalte unter 5 ppm und Eisengehalte unter 1 ppm erreicht. Der niedrige Eisengehalt ist für die Stabilität des Öles von Vorteil.It has been found that the treatment of animal or vegetable oils which have a phosphorus content of 50 to 250 ppm with a phospholipase A 1 , A 2 or B in a continuous process leads to phosphorus contents below 5 ppm if the enzyme is in the form of a treated into fine droplets of emulsified aqueous solution at a pH of 4 to 6. Phosphorus levels below 5 ppm and iron levels below 1 ppm were achieved. The low iron content is beneficial for the stability of the oil.
Die drastische Verminderung des Phosphorgehalts ist insofern überraschend, als die Phospholipasen A1, A2 und B ein pH-Optimum im neutralen bis schwach alkalischen Bereich haben. Mit Phospholipase C oder D läßt sich das Verfahrensziel nicht erreichen.The drastic reduction in the phosphorus content is surprising in that the phospholipases A 1 , A 2 and B have a pH optimum in the neutral to weakly alkaline range. The process goal cannot be achieved with phospholipase C or D.
Gegenstand der Erfindung ist daher ein Verfahren zur Verminderung des Gehaltes an phosphorhaltigen Bestandteilen in tierischen oder pflanzlichen Ölen durch enzymatischen Abbau mittels einer Phospholipase. Dabei wird ein vorentschleimtes tierisches oder pflanzliches Öl mit einem Phosphor-gehalt von 50 bis 250 ppm mit einer organischen Carbonsäure verrührt und der pH-Wert der gebildeten Mischung auf 4 bis 6 eingestellt; der Mischung wird in einem Mischgefäß eine Enzymlösung, die Phospholipase A1, A2 oder B enthält, unter turbulentem Rühren und unter Bildung feiner Tröpfchen zugegeben, wobei eine Emulsion mit 0,5 bis 5 Gew.-% Wasser, bezogen auf das Öl, gebildet wird, welche durch mindestens ein nachfolgendes Reaktionsgefäß unter turbulenter Bewegung während einer Reaktionszeit von 0,1 bis 10 Stunden bei Temperaturen im Bereich von 20 bis 80°C geleitet wird, und wobei das behandelte Öl nach Abtrennen der wäßrigen Lösung einen Phosphorgehalt unter 5 ppm aufweist.The invention therefore relates to a process for reducing the content of phosphorus-containing constituents in animal or vegetable oils by enzymatic degradation by means of a phospholipase. A pre-degummed animal or vegetable oil with a phosphorus content of 50 to 250 ppm is stirred with an organic carboxylic acid and the pH of the mixture formed is adjusted to 4 to 6; an enzyme solution containing phospholipase A 1 , A 2 or B is added to the mixture in a mixing vessel with turbulent stirring and formation of fine droplets, an emulsion containing 0.5 to 5% by weight of water, based on the oil, is formed, which is passed through at least one subsequent reaction vessel with turbulent agitation for a reaction time of 0.1 to 10 hours at temperatures in the range from 20 to 80 ° C., and the treated oil, after separating off the aqueous solution, has a phosphorus content below 5 ppm having.
Da die Phospholipasen A1, A2 oder B Lecithin angreifen würden, ist es nicht sinnvoll, hoch lecithinhaltige Öle, wie rohes Sojaöl, in dem erfindungsgemäßen Verfahren einzusetzen. Ausgangsstoff sind daher vorzugsweise vorentschleimte Öle, insbesondere Speiseöls, die sich in der Regel durch einen Phosphorgehalt zwischen 50 und 250 ppm auszeichnen. Öle schwankender Oualität können auf den gleichen Anlage verarbeitet werden. Vorzugsweise werden vorentschleimte Öle, vor allem Sonnenblumenöl; Rapsöl und insbesondere Sojaöl eingesetzt. Ein vorherige Trocknung des Öls ist entbehrlich.Since the phospholipases A 1 , A 2 or B would attack lecithin, it does not make sense to use oils with a high lecithin content, such as crude soybean oil, in the process according to the invention. The starting material is therefore preferably pre-degummed oils, especially edible oils, which are generally characterized by a phosphorus content between 50 and 250 ppm. Oils with fluctuating quality can be processed on the same system. Pre-degummed oils, especially sunflower oil; Rapeseed oil and especially soybean oil are used. It is not necessary to dry the oil beforehand.
Die Phospholipase wird zweckmäßig in wäßriger Lösung eingesetzt, die in dem Öl so fein wie möglich emulgiert wird. Die enzymatische Reaktion dürfte an der Grenzfläche zwischen der Ölphase und der Wasserphase stattfinden. Sie wird durch intensive Mischung, z.B. durch turbulentes Rühren und zusätzlich durch den Zusatz von Tensiden gefördert. Die Abbauprodukte der NHP haben eine höhere Hydrophilie und gehen daher in die Wasserphase über. Sie werden daher ebenso wie die Metallionen gleichzeitig mit der Wasserphase aus dem Öl entfernt.The phospholipase is advantageously used in an aqueous solution which is as fine as possible in the oil is emulsified. The enzymatic reaction is expected to occur at the interface between the oil phase and the Water phase take place. It is mixed intensively, e.g. through turbulent stirring and additionally promoted by the addition of surfactants. The degradation products of the NHP have a higher hydrophilicity and therefore go into the water phase. They are therefore, like the metal ions, simultaneously with the Water phase removed from the oil.
Phospholipase A1, A2 und B sind bekannte Enzyme; (Vgl. Pardun, loc.cit. Seiton 135-141). Phospholipase A1 spaltet die Fettsäureestergruppe am G1-Atom eines Phospholipidmoleküls. Sie findet sich z.B. in der Rattenleber und im Schweinepankreas. Aus Schimmelpilzkulturen von Rhizopus arrhizus konnte ein Enzym mit Phospholipase-A1-Aktivität isoliert werden.Phospholipase A 1 , A 2 and B are known enzymes; (See Pardun, loc.cit. Seiton 135-141). Phospholipase A 1 cleaves the fatty acid ester group on the G 1 atom of a phospholipid molecule. It is found, for example, in rat liver and in the pancreas. An enzyme with phospholipase A 1 activity could be isolated from mold cultures of Rhizopus arrhizus.
Phospholipase A2, die früher auch als Lecithinase A bezeichnet wurde, spaltet die Fettsäureestergruppe am C2-Atom eines Phospholipidmoleküls. Sie tritt, meist vergesellschaftet mit anderen Phospholipasen, in fast allen Tier- und Pflanzenzellen auf. Reichlich findet sie sich im Schlangengift der Klapperschlange und der Kobra, sowie im Bienen- und Skorpiongift. Technisch kann sie aus Pankreasdrüsen gewonnen werden, nachdem aktivitätsinhibierende Begleitproteine mit Trypsin abgebaut worden sind.Phospholipase A 2 , formerly also called lecithinase A, cleaves the fatty acid ester group on the C 2 atom of a phospholipid molecule. It occurs, mostly in association with other phospholipases, in almost all animal and plant cells. It is abundant in the snake venom of the rattlesnake and the cobra, as well as in the bee and scorpion venom. Technically, it can be obtained from pancreatic glands after activity-inhibiting accompanying proteins have been broken down with trypsin.
Phospholipase B kommt in der Natur weitverbreitet vor. Sie wirkt auf das durch Phospholipase A1-Einwirkung entstandene Lysolecithin durch Abspaltung des zweiten Fettsäureesterrestes ein. Zum Teil wird sie auch als Gemisch der Phospholipasen A1 und A2 angesehen. Sie kommt in der Rattenleber vor und wird auch von manchen Schimmelpilzen, wie Penicillium notatum, erzeugt.Phospholipase B is widespread in nature. It acts on the lysolecithin caused by the action of phospholipase A 1 by splitting off the second fatty acid ester residue. In some cases it is also regarded as a mixture of the phospholipases A 1 and A 2 . It occurs in rat liver and is also produced by some molds, such as Penicillium notatum.
Phospholipase A2 und B sind als Handelsprodukte erhältlich. Für die technische Anwendung ist der Einsatz gereinigter Enzyme in der Regel nicht erforderlich. Für das Verfahren der Erfindung eignet sich ein Phospholipase-Präparat, das aus gemahlenen Pankreasdrüsenbrei gewonnen wird und vor allem Phospholipase A2 enthält. Das Enzym wird - je nach Aktivität - in Mengen von 0,001 bis 1 Gew.-%, bezogen auf Öl, eingesetzt. Eine gute Verteilung des Enzyms im Öl wird gewährleistet, wenn es in einer Wassermenge von 0,5 bis 5 Gew.-%, bezogen auf Öl, gelöst und in dieser Form in dem Öl zu Tröpfchen von weniger als 10 µm Durchmesser (Gewichtsmittelwert) emulgiert wird. Turbulentes Rühren mit Radialgeschwindigkeiten über 100 cm/s hat sich bewährt. Stattdessen kann das Öl mit Hilfe einer extemen Kreiselpumpe im Reaktor umgewälzt werden. Auch mittels Ultraschalleinwirkung läßt sich die enzymatische Reaktion fördern.Phospholipase A 2 and B are available as commercial products. The use of purified enzymes is generally not necessary for technical applications. A phospholipase preparation which is obtained from ground pancreatic pulp and which contains above all phospholipase A 2 is suitable for the process of the invention. Depending on the activity, the enzyme is used in amounts of 0.001 to 1% by weight, based on oil. A good distribution of the enzyme in the oil is ensured if it is dissolved in an amount of water of 0.5 to 5% by weight, based on oil, and emulsified in this form in the oil into droplets of less than 10 μm in diameter (weight average) becomes. Turbulent stirring at radial speeds above 100 cm / s has proven itself. Instead, the oil can be circulated in the reactor using an external centrifugal pump. The enzymatic reaction can also be promoted by means of ultrasound.
Die Enzymwirkung wird durch den Zusatz einer Säure, vorzugsweise einer organischen Carbonsäure, gesteigert, die vor der Enzymbehandlung zugegeben wird, Citronensäure ist bevorzugt. Sie kann in Form der freien Säure oder als Puffersystem in Kombination mit ihrem, Salz, wie einem Alkalisalz (z.B. Natriumcitrat), einem Erdalkalisalz (z.B. Calciumcitrat) oder einem Ammoniumsalz, eingesetzt werden. Geeignete Mengen sind 0,01 - 1 Gew.-%, bez. auf Öl, optimal 0,1 Gew.-%. Mit der Säure wird der pH-Wert auf einen Wert von 3 bis 7, vorzugsweise von 4 bis 6, eingestellt. Das Optimum liegt etwa bei pH 5: Überraschenderweise ist dieser pH-Wert auch dann optimal, wenn die Phospholipase in Form eines Enzymkomplexes aus Pankreas eingesetzt wird. Der Pankreas-Enzymkomplex hat sonst ein pH-Optimum von 8 und ist bei pH 5 kaum noch wirksam ist. Anscheinend stellt sich an der Phasengrenzfläche, wo die Enzymwirkung eintritt, ein höherer pH-Wert als innerhalb der wäßrigen Phase ein. The enzyme effect is achieved by adding an acid, preferably an organic carboxylic acid, increased, which is added before the enzyme treatment, Citric acid is preferred. It can be in the form of the free acid or in combination as a buffer system with their salt, such as an alkali salt (e.g. sodium citrate), an alkaline earth salt (e.g. calcium citrate) or a Ammonium salt can be used. Suitable amounts are 0.01-1% by weight, based on on oil, optimally 0.1% by weight. The pH is adjusted to a value of 3 to 7, preferably 4 to 6, with the acid. The The optimum is around pH 5: Surprisingly, this pH is also optimal when the Phospholipase is used in the form of an enzyme complex from the pancreas. The pancreatic enzyme complex otherwise has a pH optimum of 8 and is hardly effective at pH 5. Apparently it turns out to be Phase interface where the enzyme action occurs, a higher pH than within the aqueous phase a.
Um die Phospholipasen A1, A2 und B aus fetthältigem Pankreatin oder Pankreasprodukten in Lösung zu bringen, sind emulgierende Zusätze hilfreich. Geeignet sind wasserlösliche Emulgatoren, insbesondere solche mit einem HLB-Wert über 9, wie Na-Dodecylsulfat. Es ist in einer Menge von z.B. 0,001 Gew.-%, bezogen auf Öl, wirksam, wenn es der Enzymlösung vor dem Emulgieren im Öl zugesetzt wird.In order to dissolve the phospholipases A 1 , A 2 and B from fatty pancreatin or pancreatic products, emulsifying additives are helpful. Water-soluble emulsifiers, in particular those with an HLB value above 9, such as sodium dodecyl sulfate, are suitable. It is effective in an amount of, for example, 0.001% by weight based on oil if it is added to the enzyme solution prior to emulsification in the oil.
Der Zusatz weiterer Enzyme, vor allem Proteinasen und Amylasen, wirkt sich oft vorteilhaft aus. Auch Proteinzusätze können durch eine gewisse Tensidwirkung vorteilhaft sein.The addition of other enzymes, especially proteinases and amylases, often has an advantageous effect. Also Protein additives can be advantageous due to a certain surfactant effect.
Die Temperatur bei der Enzymbehandlung ist nicht kritisch. Temperaturen zwischen 20 und 80°C sind geeignet. Optimal ist eine Temperatur von 50°C, jedoch kann kurzzeitig auch bis 70°C erwärmt werden. Die Behandlungsdauer hängt von der Temperatur ab und kann mit zunehmender Temperatur kürzer gehalten werden. Zeiten von 0,1 bis 10, vorzugsweise 1 bis 5 Stunden sind in der Regel ausreichend. Besonders bewährt hat sich ein Stufenprogramm, bei dem die erste Stufe bei einer Temperatur von 40 bis 60°C und die zweite Stufe bei einer höheren Temperatur im Bereich von 50 bis 80°C ausgeführt wird. Beispielsweise wird zuerst 3 Stunden bei 50°C, dann eine Stunde bei 75°C gerührt.The temperature in the enzyme treatment is not critical. Temperatures are between 20 and 80 ° C suitable. A temperature of 50 ° C is optimal, but it can also be briefly heated to 70 ° C. The duration of treatment depends on the temperature and can be shorter with increasing temperature being held. Times of 0.1 to 10, preferably 1 to 5 hours are usually sufficient. A step program, in which the first step at a temperature of 40 to 60 ° C and the second stage is carried out at a higher temperature in the range of 50 to 80 ° C. For example, the mixture is first stirred at 50 ° C. for 3 hours, then at 75 ° C. for one hour.
Nach Abschluß der Behandlung wird die Enzymlösung mitsamt der darin aufgenommenen Abbauprodukte der NHP von der Ölphase abgetrennt, vorzugsweise durch Zentrifugieren. Da sich die Enzyme durch eine hohe Stabilität auszeichnen und die Menge der aufgenommenen Abbauprodukte gering ist, kann die gleiche Enzymlösung mehrmals wiederverwendet werden.After completion of the treatment, the enzyme solution together with the degradation products contained therein the NHP is separated from the oil phase, preferably by centrifugation. Because the enzymes get through a high stability and the amount of degradation products absorbed is low same enzyme solution can be reused several times.
Vorzugsweise wird das Verfahren kontinuierlich durchgeführt. Bei einer zweckmäßigen kontinuierlicher Arbeitsweise wird das Öl in einem ersten Mischgefäß mit der Enzymlösung emulgiert, in einem oder mehreren nachfolgenden Reaktionsgefäßen, gegebenenfalls bei steigender Temperatur, unter turbulenter Bewegung reagieren gelassen, und anschließend in einer Zentrifuge die wäßrige Enzymlösung abgetrennt. Um eine Anreicherung der Abbauprodukte in der Enzymlösung zu vermeiden, kann ein Teil davon laufend durch frische Enzymlösung ersetzt und der Rest in den Prozeß zurückgeführt werden.The method is preferably carried out continuously. With an expedient continuous The oil is emulsified in a first mixing vessel with the enzyme solution, in one or several subsequent reaction vessels, if necessary with increasing temperature, under turbulent Movement allowed to react, and then the aqueous enzyme solution separated in a centrifuge. To avoid an accumulation of the degradation products in the enzyme solution, a part of it can be running replaced with fresh enzyme solution and the rest returned to the process.
Das gewonnene Öl hat einen Phosphor-Gehalt unter 5 ppm und ist damit zur physikalischen Raffination zu Speiseöl geeignet. Dank des erreichten niedrigen Eisengehaltes hat es gute Voraussetzungen, um beim Raffinieren eine hohe Oxydationsbeständigkeit zu erreichen.The oil obtained has a phosphorus content below 5 ppm and is therefore for physical refining suitable for cooking oil. Thanks to the low iron content that has been achieved, it has good preconditions for Refining to achieve high resistance to oxidation.
1 l naßentschleimtes Sojaöl mit einem Rest-Phosphorgehalt von 110 ppm wird in einem Rundkolben auf
75°C erwärmt. Unter starkem Rühren mit einem Flügelrührer von 5 cm Durchmesser bei 700 UpM werden
10 ml Wasser, enthaltend 1 g Citronensäure, zugesetzt und 1 Stunde weitergerührt. Dann wird auf 40°C
abgekühlt und eine Lösung von 0,1 g Phospholipase A2 der in Beispiel 1 angegebenen Qualität sowie 50
mg Calciumchlorid in 20 ml 0,1 molarem Acetatpuffer pH 5,5 zugegeben. Nach weiteren 5 Stunden
intensiver Rührung wird die Wasserphase abzentrifugiert. Das erhaltene Öl enthält 2 ppm P und ist zur
physikalischen Raffination geeignet. Die Veränderung der anderen Kennzahlen geht aus folgender Tabelle
hervor:
Das Verfahren gemäß Beispiel 1 wird mit dem Unterschied wiederholt, daß anstelle von Phospholipase A2 1 g eines Pankreaspräparats (Pankreatin, 800 Phospholipase-Einheiten/g) eingesetzt wird. Das Präparat enthält Phospholipase A2, Proteinase, Amylase, Lipase. Der Phosphorgehalt sinkt unter 1 ppm. Durch den Einfluß der Lipase tritt nur ein geringfügiger Anstieg der Säurezahl von 0,91 auf 1,49 ein.The procedure according to Example 1 is repeated with the difference that 1 g of a pancreatic preparation (pancreatin, 800 phospholipase units / g) is used instead of phospholipase A 2 . The preparation contains phospholipase A 2 , proteinase, amylase, lipase. The phosphorus content drops below 1 ppm. Due to the influence of lipase, there is only a slight increase in the acid number from 0.91 to 1.49.
9 l naßentschleimtes Rapsöl mit einem Phosphorgehalt von 72 ppm wird mit einer Lösung von 8.6 g Citronensäure in 250 ml Wasser vermischt und auf 60°C erhitzt. Die Mischung wird homogenisiert, indem man sie mittels einer externen Kreiselpumpe einmal pro Minute umwälzt. Dann wird der pH-Wert der wäßrigen Phase mit 30 g 10 %iger Natronlauge auf 5,0 eingestellt. Nun werden 9 g Phospholipase A2 mit einer Aktivität von 400 U/g zusammen mit etwas Calciumchlorid zugesetzt und die Mischung 3 Stunden bei 60°C in der angegebenen Weise umgewälzt. Nach dem Abzentrifugieren wird ein Phosphorgehalt von 3 ppm gefunden. 9 l wet-degummed rapeseed oil with a phosphorus content of 72 ppm is mixed with a solution of 8.6 g citric acid in 250 ml water and heated to 60 ° C. The mixture is homogenized by circulating it once per minute using an external centrifugal pump. Then the pH of the aqueous phase is adjusted to 5.0 with 30 g of 10% sodium hydroxide solution. Now 9 g of phospholipase A 2 with an activity of 400 U / g are added together with a little calcium chloride and the mixture is circulated for 3 hours at 60 ° C. in the manner indicated. After centrifugation, a phosphorus content of 3 ppm is found.
Claims (4)
- A process for reducing the content of phosphorus-containing ingredients in animal or vegetable oils through enzymatic degradation using a phospholipase, in doing so a previously-deslimed animal or vegetable oil with a phosphorus content of 50 to 250 ppm is stirred with an organic carbonic acid and the pH value of the resultant mixture is set at 4 to 6; an enzyme solution containing phospholipase A1, A2 or B is added to the mixture in a mixing vessel, whilst being subjected to turbulent stirring with the formation of fine droplets, wherein an emulsion is formed with 0.5 to 5 wt. % water, relating to the oil, which is passed through at least one subsequent reaction vessel, undergoing turbulent stirring, during a reaction time of 0.1 to 10 hours at temperatures in the range of 20° to 80° C, and wherein the treated oil has a phosphorus content of less than 5 ppm after separation of the aqueous solution.
- A process according to claim 1, characterised in that an oil is used from which mucilage has been removed beforehand.
- A process according to claim 1 or 2, characterised in that a citric acid is used as an organic carbonic acid in the form of the free acid or as an alkali metal, calcium or ammonium citrate.
- A process according to one or more of claims 1 to 3, characterised in that an emulsifier is additionally used.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE4115938A DE4115938A1 (en) | 1991-05-16 | 1991-05-16 | ENZYMATIC METHOD FOR REDUCING THE CONTENT OF PHOSPHORUS-CONTAINING COMPONENTS IN VEGETABLE AND ANIMAL OILS |
DE4115938 | 1991-05-16 |
Publications (4)
Publication Number | Publication Date |
---|---|
EP0513709A2 EP0513709A2 (en) | 1992-11-19 |
EP0513709A3 EP0513709A3 (en) | 1992-12-30 |
EP0513709B1 EP0513709B1 (en) | 1995-03-29 |
EP0513709B2 true EP0513709B2 (en) | 1999-10-06 |
Family
ID=6431742
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP92107888A Expired - Lifetime EP0513709B2 (en) | 1991-05-16 | 1992-05-11 | Enzymatic method for reducing the amount of phosphorous-containing components in vegetable and animal oils |
Country Status (15)
Country | Link |
---|---|
US (1) | US5264367A (en) |
EP (1) | EP0513709B2 (en) |
CN (1) | CN1034587C (en) |
AR (1) | AR245193A1 (en) |
AT (1) | ATE120482T1 (en) |
BR (1) | BR9201859A (en) |
CA (1) | CA2068933C (en) |
DE (2) | DE4115938A1 (en) |
DK (1) | DK0513709T4 (en) |
ES (1) | ES2072043T5 (en) |
GR (2) | GR3015920T3 (en) |
HU (1) | HU213754B (en) |
PL (1) | PL170548B1 (en) |
RU (1) | RU2033422C1 (en) |
TW (1) | TW203625B (en) |
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Family Cites Families (14)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
FR70269E (en) * | 1956-06-18 | 1959-03-25 | Circuit-breaker device for hydraulic circuits | |
US3522145A (en) * | 1966-07-20 | 1970-07-28 | Colgate Palmolive Co | Deodorization of fats |
GB1440462A (en) * | 1973-06-29 | 1976-06-23 | Stork Amsterdam | Process for the clarification and/or recovery of vegetable oils |
LU83441A1 (en) * | 1981-06-19 | 1983-04-06 | Tirtiaux Sa | PROCESS FOR TREATING OILS AND FATS AND PRODUCTS THUS OBTAINED |
US4420560A (en) * | 1981-11-17 | 1983-12-13 | Fuji Oil Company, Limited | Method for modification of fats and oils |
US4478940A (en) * | 1981-12-24 | 1984-10-23 | Novo Industri A/S | Production of purified vegetable protein |
US4483874A (en) * | 1982-05-06 | 1984-11-20 | Novo Industri A/S | Preparation of milk substitute |
GB8506907D0 (en) * | 1985-03-18 | 1985-04-24 | Safinco Coordination Centre Nv | Removal of non-hydratable phoshatides from vegetable oils |
GB8604133D0 (en) * | 1986-02-19 | 1986-03-26 | Unilever Plc | Spreads |
EP0328789B1 (en) * | 1988-02-18 | 1992-04-08 | Unilever N.V. | Heat-sterilizable water and oil emulsion |
JP2794574B2 (en) * | 1988-08-11 | 1998-09-10 | 昭和産業株式会社 | Method for producing lysolecithin |
JP2709736B2 (en) * | 1988-08-11 | 1998-02-04 | 昭和産業株式会社 | Oil and fat refining method |
JP2701387B2 (en) * | 1988-12-06 | 1998-01-21 | 三菱化学株式会社 | Recording liquid |
JPH0738771B2 (en) * | 1989-01-17 | 1995-05-01 | 花王株式会社 | Liquid edible oil composition |
-
1991
- 1991-05-16 DE DE4115938A patent/DE4115938A1/en not_active Withdrawn
-
1992
- 1992-05-11 DE DE59201753T patent/DE59201753D1/en not_active Expired - Lifetime
- 1992-05-11 ES ES92107888T patent/ES2072043T5/en not_active Expired - Lifetime
- 1992-05-11 DK DK92107888T patent/DK0513709T4/en active
- 1992-05-11 EP EP92107888A patent/EP0513709B2/en not_active Expired - Lifetime
- 1992-05-11 AT AT92107888T patent/ATE120482T1/en active
- 1992-05-12 CN CN92103459A patent/CN1034587C/en not_active Expired - Lifetime
- 1992-05-14 PL PL92294543A patent/PL170548B1/en unknown
- 1992-05-14 RU SU925011752A patent/RU2033422C1/en active
- 1992-05-14 US US07/882,710 patent/US5264367A/en not_active Expired - Lifetime
- 1992-05-15 TW TW081103810A patent/TW203625B/zh not_active IP Right Cessation
- 1992-05-15 BR BR929201859A patent/BR9201859A/en not_active Application Discontinuation
- 1992-05-15 HU HU9201630A patent/HU213754B/en unknown
- 1992-05-15 AR AR92322337A patent/AR245193A1/en active
- 1992-05-19 CA CA002068933A patent/CA2068933C/en not_active Expired - Lifetime
-
1995
- 1995-04-25 GR GR950401041T patent/GR3015920T3/en unknown
-
1999
- 1999-11-10 GR GR990402897T patent/GR3031804T3/en unknown
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Also Published As
Publication number | Publication date |
---|---|
BR9201859A (en) | 1993-01-05 |
ATE120482T1 (en) | 1995-04-15 |
EP0513709B1 (en) | 1995-03-29 |
PL170548B1 (en) | 1996-12-31 |
DK0513709T3 (en) | 1995-07-24 |
GR3015920T3 (en) | 1995-07-31 |
RU2033422C1 (en) | 1995-04-20 |
ES2072043T3 (en) | 1995-07-01 |
CA2068933A1 (en) | 1992-11-17 |
TW203625B (en) | 1993-04-11 |
GR3031804T3 (en) | 2000-02-29 |
DE59201753D1 (en) | 1995-05-04 |
HU213754B (en) | 1997-09-29 |
AR245193A1 (en) | 1993-12-30 |
CA2068933C (en) | 1995-12-19 |
ES2072043T5 (en) | 2000-02-01 |
DK0513709T4 (en) | 1999-12-27 |
CN1034587C (en) | 1997-04-16 |
CN1066679A (en) | 1992-12-02 |
EP0513709A3 (en) | 1992-12-30 |
DE4115938A1 (en) | 1992-11-19 |
PL294543A1 (en) | 1993-01-25 |
HU9201630D0 (en) | 1992-08-28 |
US5264367A (en) | 1993-11-23 |
HUT64578A (en) | 1994-01-28 |
EP0513709A2 (en) | 1992-11-19 |
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