Classifications

• • C—CHEMISTRY; METALLURGY
  • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
  • C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
  • C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
  • C11D3/16—Organic compounds
  • C11D3/38—Products with no well-defined composition, e.g. natural products
  • C11D3/386—Preparations containing enzymes, e.g. protease or amylase
  • C11D3/38627—Preparations containing enzymes, e.g. protease or amylase containing lipase
• • 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
  • C11B1/00—Production of fats or fatty oils from raw materials
  • C11B1/02—Pretreatment
  • C11B1/025—Pretreatment by enzymes or microorganisms, living or dead

Definitions

• PROCESS FOR ENZYMATIC DEGREASING OF BONES BACKGROUND OF THE INVENTION 1.
• This invention relates to a method for the degreasing of 5 bones and, in particular, concerns a method whereby the bones are subjected to the action of lipolytic enzymes (lipases).
• Bones from which gelatin is to be extracted must first be degreased. When bones are naturally degreased due to their exposure to the atmosphere and in particular the action of sun and rain, there is usually no need for further treatment prior to the procedure for the extraction of gelatin. Bones which come directly from the slaughter houses contain a high lipoid content which must be considerably reduced before an efficient gelatin extraction procedure can be commenced.
• the present invention is based on the discovery that bones, particularly those having a lipoid content in the range from about to 35 percent by weight, based upon bones containing 10 percent water, when treated with an aqueous solution of lipolytic enzymes (lipases), in the presence of a soluble calcium salt, effectively degrease the bones when they are contacted with such a solution at a temperature in the range from about to 40 C.
• lipolytic enzymes lipases
• the lipases useful in this invention may be of miscellaneous origin such as animal, vegetable or microbial.
• the animal lipases are generally of pancreatic origin and are unstable and the pancreatic lipases contain proteolytic enzymes which may attach the bones protein.
• Vegetable lipases present problems mainly due to the difficulty with which they are obtained.
• lipases of microbiological origin are best suitable for use in the method of this invention. These lipases can be efficiently produced under normal industrial conditions and they have the ability to hydrolyze the triglycerides of long chain fatty acids which occur in bone lipoids and to transform these materials into diglycerides, monoglycerides, glycerol and fatty acids.
• the quantity and type of lipase to be used will depend upon the origin of the bones, their lipoid content and the particular properties of the lipase.
• the quantity of calcium salt to be used will depend upon the quantity of bones to be treated since there is in each case an optimum proportion with respect to the quantities of lipoid, lipase and soluble calcium salt. However, an excess of calcium is not harmful to the reaction and can be used in the process if necessary.
• the fungic lipases are preferred and, in particular, the lipase of Rhizopus arrhizus var. Delemar (Ra.D.) because it is very stable, it has an optimum pH and an optimum temperature and it has a mode of action which makes it particularly efficient for the degreasing of bones.
• Other lipases which are efficiently used in this process are the lipases of Rhizopus nigricans, Aspergillus niger and Candida cylindracea.
• the enzymatic solution is separated from the bones which are washed with water, preferably hot, to eliminate the sticking products of the reaction which would tend to inhibit the action of the lipase during subsequent treatments.
• the bones which have been subjected to the degreasing treatment described hereinabove are ready for further treatment, particularly the extraction of gelatin.
• the bones to be degreased should be treated at a pH in the range from between about 5 to about 8, the temperature should be maintained in the range from between about 20 to about 40 C. and the treatment, which may vary in this process anywhere from 3 to 20 hours, will depend upon the lipoid content in the bones and the activity of the particular lipase used in the process. For example, when lipase R.a.D. is used, the treatment is continued for a period of from between 6 to 12 hours for an enzyme titrating about 10,000 ;/./g. specific activity. For a definition of this term see the reference above cited.
• Bone pieces which are able to pass through a sieve having square meshes of 16 mm. or 19 mm. on a side are preferred.
• the previously crushed bones (called green bones"), be subjected to a pretreatment with hot water for a period of time ranging from about 15 to minutes. This results in the washing out of part of the lipoids which adhere to the bones while some residual matter remains.
• the pretreatment may be repeated if desired.
• Each kilogram of bones to be degreased is treated with an aqueous solution of from 0.2 to 10 g. of lipase, the volume of the lipase generally ranging from between 0.5 to 3.0 liters per kilogram of bones.
• a quantity of soluble calcium salt, preferably calcium chloride, is added to the lipase solution in an amount ranging from 0.2 to 10 g. of calcium salt per kilogram bones.
• the calcium aids in the elimination of fatty acids resulting from the action of the lipase by combining with these acids and eliminating them from the reaction medium which thus enhances the action of the enzyme; it forms a part of the enzyme molecule and acts as a specific promoter of the lipase; and it protects the enzyme and improves the effectiveness and economy of the process.
• the bones are gently stirred, either continuously or intermittently, in the lipase solution, thus effecting maximum contact between the lipase solution and the bones.
• the enzymatic solution is separated from the bones which are then washed with hot water to eliminate the reaction products which otherwise would tend to inhibit the action of the lipase in further treatment.
• the washing which may be carried out at a pH of from between 7 to 8.5. and preferably between pH 8 and pH 8.5, may be repeated as many times as necessary.
• EXAMPLE 1 Slaughterhouse bones having a lipoid content of 23.2 percent (based on bones containing percent water) were crushed and passed through a 19 mm. mesh sieve. The bones were washed in boiling water for 30 minutes, after which the liquid containing lipoids and meat parts was eliminated and the operation repeated.
• One kilogram of the bones was introduced into 1 liter of an aqueous solution of lipase R.a.D. containing 0.5 g./l. of lipase (activity 9,000 a/g.) together with 3 g. of CaCl
• the treatment was continued for 6 hours at 37 C. while the solution was continuously stirred.
• the pH was continuously maintained at 7.2102 by adding an aqueous solution of 0.5 M NaOH.
• the solution was drawn off and the bones were stirred in water at 75 C. for minutes, after which another lipasic treatment as above was commenced, using 0.5 g. of lipase, (activity 9,000 p/g.) together with 9 g. of CaCl per kilogram of bones.
• the third and last treatment with lipase was followed by a last water washing at 75 C.
• EXAMPLE 2 Slaughterhouse bones having a lipoid content of 19 percent (based on bones containing 10 percent water) were treated.
• the bones were constituted as follows: half of the pieces being about 16 mm. in size and half being about 19 mm. They were washed in boiling water for 40 minutes, after which the solution containing lipoids and meat remains was eliminated.
• the solution was drawn off and the bones stirred with water at 70 C. for 20 minutes, the water having been brought to and then maintained at pH 8.5 by the addition of a dilute aqueous solution of NaOH.
• a second treatment was commenced with a new aqueous solution of lipase R.a.D. containing 2.5 g. of lipase (activity 7,500 p/g. per kilogram of bones, i.e., about 18,000 pi/kg. of bones) and 9 g./l. of CaCl the quantities of solutions being the same comparatively to the weight of bones as in the first treatment.
• the second treatment was continued for 12 hours at 37 C., the pH maintained, as in the first treatment, at 72:02 by a solution of 0.5 M NaOH.
• the treatment was continued for 8 hours at 37 C., the pH being maintained at 7.2i0.2 by the addition of 0.5 M KOH.
• the lipasic solution was drawn off and the bones stirred for 30 minutes with water brought to C., the pH of which was maintained at 8.4 by the addition of a dilute solution of KOH.
• the alkaline rinsing solution was eliminated and a second rinsing commenced with water at 75 C., with vigorous stirring.
• a second lipasic treatment was commenced with 2,000 liters of a lipasic solution containing 1 g./l. of lipase of R.a.D. (activity 12,000 p/g, i.e., 12,000 units per kilogram of bones) and 6 g./l. of CaCI the treatment was continued for 10 hours at 37 C., while maintaining the pH at 7.2i0.2 by the addition of an aqueous solution of 0.5 M KOH.
• the solution was drawn OB and the bones were rinsed by stirring in water at 75 C. for 40 minutes, the pH being brought to and maintained at 8.5 by the addition of an aqueous solution of dilute NaOH.
• a second treatment was then commenced identical to the first, and continuing for 15 hours, after which the bones were rinsed under the same conditions as above.
• One thousand kilograms of the treated bones were placed into 10,000 liters of an aqueous solution of lipase of Rhizopus nigricans containing 1 g./l. of enzyme (activity 5,200 p./g., i.e., 5,200 .1. per kilogram of bones) and 3 g./l. of CaCl
• the treatment was continued for 8 hours at 28 C., the pH being maintained at 6.9102, this pH being an average between the optimum pH of the enzyme and the pl( of the fatty acids.
• a second treatment was commenced with 10,000 liters of a solution of lipase containing 1 g./l. of lipase of Rhizopus nigricans, (activity 5,200 u/g, i.e., 10,400 1. per kilogram of bones) and 3 g./l. of CaCl
• the treatment was continued for 10 hours at 28 C. under the same conditions as in the first treatment, followed by a rinsing, as previously done.
• EXAMPLE 6 Crushed slaughterhouse bones of 19 mm. size and having a lipoid content of 23.4 percent based on bones containing 10 percent water were subjected to the action of boiling water for 45 minutes. After treatment the liquid was eliminated and an identical treatment commenced.
• the solution of lipase was drawn 011' after which 50 liters of water brought to C. was added the pH of which was maintained at 8.5 by the addition 0 dilute NaOH, accompanied by stirring for 15 minutes; the solution was then eliminated, and a second rinsing under the same conditions commenced.
• a second lipasic treatment was commenced with 759 liters of an aqueous solution containing 2.1 g./l. of lipase (i.e., 7,700p. per kilogram of bones), and 2 g./l. of CaCl
• the treatment was continued with discontinuous stirring for 15 hours at 30 C., the pH being maintained at 7.2i0.2 by the addition of a solution of a 0.5 M NaOH. This treatment was followed by a rinsing.
• a method for the enzymatic degreasing of bones which comprises subjecting said bones to the action of a lipolytic enzyme solution in the presence of a soluble calcium salt.
• said lipolytic enzyme solution is at a pH in the range from about 5 to about 8 and at a temperature in the range from about 20 to about 45 C.
• a method according to claim 2 wherein said lipolytic enzyme is of microbiological origin.
• said lipolytic en zyme is a member of the group consisting of the lipases of Rhizopus arrhizus Delemar, Rhizopus nigricans, Aspergillus niger, Candida cylindracea, Mucor mucedo, Rhizpus oryzae, Candida lipolytica and Aspergillus Iucheunsis.
• said soluble calcium salt is calcium chloride present at a concentration of from 0.2 g. to 10 g. per kilogram of bones.

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• Chemical & Material Sciences (AREA)
• Life Sciences & Earth Sciences (AREA)
• Engineering & Computer Science (AREA)
• Chemical Kinetics & Catalysis (AREA)
• Oil, Petroleum & Natural Gas (AREA)
• Wood Science & Technology (AREA)
• Organic Chemistry (AREA)
• Biochemistry (AREA)
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• Preparation Of Compounds By Using Micro-Organisms (AREA)
• Meat, Egg Or Seafood Products (AREA)

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Cited By (4)

Citations (1)

• 1967
  • 1967-10-20 FR FR125196A patent/FR1547744A/fr not_active Expired
• 1968
  • 1968-10-10 DK DK487368AA patent/DK127436B/da unknown
  • 1968-10-18 NL NL686814952A patent/NL148095B/xx unknown
  • 1968-10-18 GB GB49616/68A patent/GB1242105A/en not_active Expired
  • 1968-10-21 US US776304A patent/US3634191A/en not_active Expired - Lifetime

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