WO2014189356A1 - Procédés pour extraire des protéines de tourteaux de palmistes ou pour augmenter la teneur en protéines de tourteaux de palmistes - Google Patents

Procédés pour extraire des protéines de tourteaux de palmistes ou pour augmenter la teneur en protéines de tourteaux de palmistes Download PDF

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Publication number
WO2014189356A1
WO2014189356A1 PCT/MY2014/000074 MY2014000074W WO2014189356A1 WO 2014189356 A1 WO2014189356 A1 WO 2014189356A1 MY 2014000074 W MY2014000074 W MY 2014000074W WO 2014189356 A1 WO2014189356 A1 WO 2014189356A1
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Prior art keywords
pkc
process according
protein
enzymes
hydrolysate
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PCT/MY2014/000074
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English (en)
Inventor
Julia BINTI IBRAHIM
Shwu Fun KUA
Norhafizi BIN HASHIM
Khairul Izwan BIN NAN
Christopher Kuok Weng OOI
Hirzun MOHD YUSOF
Harikrishna Kulaveerasingam
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Sime Darby Malaysia Berhad
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Publication of WO2014189356A1 publication Critical patent/WO2014189356A1/fr

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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
    • A23J3/00—Working-up of proteins for foodstuffs
    • A23J3/14—Vegetable proteins
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
    • A23J3/00—Working-up of proteins for foodstuffs
    • A23J3/30—Working-up of proteins for foodstuffs by hydrolysis
    • A23J3/32—Working-up of proteins for foodstuffs by hydrolysis using chemical agents
    • A23J3/34—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes

Definitions

  • the present disclosure relates generally to processes for the treatment of palm kernel cake. More particularly, the disclosure relates to processes for the enzyme catalysed hydrolysis of palm kernel cake and to products obtained thereby.
  • the fruit of the oil palm Elaeis guineensis, yields two distinct oils: palm oil, derived from the outer parts of the fruit, and palm kernel oil, derived from the seed of the fruit, or kernel.
  • the palm kernel contains a hypothetical oil content of approximately 50-55%.
  • high pressure expellers are used to extract the oil from the kernel, although solvent-based extraction processes can also be used.
  • the extraction process affords the palm kernel oil and, as the waste by-product, the "de-oiled" solid mass, or pulp, referred to as "palm kernel cake” (PKC), or sometimes, palm kernel expeller” (PKE).
  • the PKC While the extraction process affords the majority of the oil present in the kernel a significant proportion of the oil is retained in the PKC (about 10% w/w).
  • the PKC also contains some protein and significant amounts of polysaccharides, including hemicelluloses, celluloses and pectins which make up the cell walls, as well as a high phosphorus to calcium ratio. PKC is therefore utilised as a high energy, high fibre feed supplement for ruminants.
  • a relatively low nutrition value and only average protein content, together with a poor amino acid profile means that PKC, on its own, is ever only a medium quality feed, ranked a little higher than copra cake but lower than fish meal and ground nut cake in its protein value.
  • PKC can be considered as a low value product.
  • oilseed proteins are potentially important sources of human and animal dietary protein and approximately 1.4 million tons of PKC-by product are produced in Malaysia alone each year. Therefore, a need remains for processes which may allow for recovery of commercially useful components, such as proteins, from the PKC by-product, and processes which increase the protein content of PKC to improve its nutritional value as an animal feedstuff.
  • the present disclosure describes processes which utilise an enzyme-catalysed hydrolysis of PKC.
  • the hydrolytic process employs one or more enzymes which advantageously breakdown, or degrade, the polysaccharide and, optionally protein components which comprise plant cell walls, releasing retained oil, smaller mono-, di- and oligosaccharides, as well as protein - commercially useful components of PKC.
  • the hydrolysis process may therefore be used to recover one or more of these components, in particular protein.
  • the processes may also afford a PKC product with a relatively increased protein content. Such extracted protein and PKC product, may find use in products for human or animal consumption.
  • the present disclosure provides a process for extracting protein from palm kernel cake (PKC) comprising:
  • the present disclosure provides a process for producing high protein content palm kernel cake (PKC) comprising:
  • the PKC subjected to hydrolysis is expeller pressed, derived from the mechanical screw press oil extraction method. In some embodiments, the PKC is derived from solvent extraction methods.
  • the enzymes are dissolved or suspended in water to provide an enzymatic solution which is mixed with the PKC.
  • one or more enzymes capable of hydrolysing at least a portion of hemicellulose polysaccharides present in the PKC are used.
  • one or more enzymes capable of hydrolysing or degrading at least a portion of mannan present in the PKC are used.
  • the process utlizes one or more enzymes capable of hydrolysing or degrading other cell wall constituents such as celluloses, pectins and associated proteins.
  • the enzymes used include one or more of mannanases, cellulases, pectinases and optionally proteinases.
  • the process employs at least mannanase and one or more enzymes selected from cellulases, pectinases and optionally proteinases.
  • the enzymes used include mannanase and pectinase.
  • the PKC hydrolysate obtained from the hydrolysis process is further subjected to additional steps prior to precipitation of the protein. These steps may include one or more of separation or clarification to remove any non-aqueous components, i.e. palm kernel oil, and filtration, such as crossflow (tangential) ultrafiltration thereby increasing the ratio of protein to non-protein components to form a protein-containing concentrate.
  • a protein-containing concentrate contains proteins having a molecular weight in the range of about 2.5 kda to about 116 kda.
  • the protein is precipitated using an antichaotropic salt, such as sodium sulphate, sodium chloride or ammonium sulphate.
  • an antichaotropic organic solvent such as ethanol.
  • the PKC produced has a protein content of at least about 20%. In further embodiments, the PKC has a protein content of at least about 25% or about 30%.
  • PKC having a protein content of at least about 20%.
  • the PKC has a protein content of at least about 25% or about 30%.
  • Figure 1 schematically depicts one example of a process for the recovery of protein from PKC.
  • Figure 2 is a ID-SDS gel image depicting bands of protein in the range of about 30-115 kda in both crude hydrolysate and retentate obtained by ultrafiltration.
  • Figure 3 graphically depicts the amount of protein precipitated (mg/g PKC) using each of sodium sulphate, sodium chloride and ammonium sulphate.
  • Figure 4 graphically depicts the amount of protein precipitated (mg/g PKC) using ammonium sulphate at concentrations of 50, 60 and 65 w/v%.
  • Figure 5 graphically depicts the amount of protein precipitated (mg/g PKC) using TCA- acetone and ammonium sulphate.
  • Figure 6 graphically depicts the amount of protein precipitated (mg/g P C) using varying concentrations, 10-100%(v/v), of absolute ethanol
  • Figure 7 graphically depicts the amount of protein precipitated (mg g PKC) using varying concentrations of absolute ethanol at 4, 8 and 25°C.
  • Figure 8 is a 1D-SDS gel image depicting bands of protein recovered by precipitation with absolute ethanol.
  • Figure 9 schematically depicts one non-limiting example of a process for the preparation of high protein content PKC.
  • invention includes all aspects, embodiments and examples described herein.
  • Palm kernel cake is the material retained after extraction of palm kernel oil from the palm kernel.
  • Two major processes are typically used; a high pressure screw press method (e.g. expeller press) or a solvent-based (e.g. hexane) extraction method.
  • the PKC which is produced by each method differs in the amount of palm kernel oil retained in the cake : 5- 12% for expeller pressed PKC versus 0.5-3% for solvent extracted PKC.
  • the amount of crude protein content for each type is comparable: about 14-16%.
  • the PKC subjected to hydrolysis is that obtained after the first pressing of palm kernels through high pressure expeller press (first pressed PKC).
  • the PKC subjected to hydrolysis is that obtained after the second pressing of palm kernels through high pressure expeller press (second pressed PKC).
  • the PKC is obtained from solvent-based (e.g. hexane) extraction of palm kernel oil. It will be understood that the PKC used in the processes described herein may be PKC obtained from one or more such processes.
  • Reference to PKC includes the material obtained directly from the oil extraction process.
  • the material may be subjected to one or more further physical and/or chemical treatments prior to being subjected to enzymatic hydrolysis.
  • Such treatments may include one or more of milling, crushing, grinding, chopping or other process of communition, hydrothermal treatment, such as steam or hot liquid water treatment, and acid (e.g. sulphuric acid) or base (e.g. sodium hydroxide) treatment.
  • Pre-treating the PKC chemically or physically helps to alter the hemicellulose structure of the PKC. This causes the PKC to be more accessible to the enzymes.
  • the PKC subjected to enzymatic treatment may be in the form of meal, flakes, granules, pellets or any other suitable form.
  • the PKC has a size in the range of about mesh size 1mm to about mesh size 10mm, for example from about l-5mm.
  • Hemicelluloses are a heterogeneous group of polysaccharides, which together with celluloses, pectins, and also proteins, are present in plant cell walls.
  • the group of polysaccharides known as hemicelluloses include polysaccharides having -(l ⁇ 4)-linked backbones of glucose, mannose or xylose with an equatorial configuration, and include xyloglucans, xylans, mannans (including galactomannans) and glucomannans (including galactoglucomannans).
  • Xylans feature a backbone of P-(l ⁇ 4)-linked xylose residues
  • mannans feature a backbone predominantly of P-(l ⁇ 4)-linked mannose units
  • glucomannans contain a backbone of glucose and mannose
  • xyloglucans contain a backbone of glucose with xylose-containing sidechains (Scheller, H.V. and Ulvskov, P., Annu. Rev. Plant Biol.2010, 61 , 263-289).
  • the processes described herein subject P C to enzymatic hydrolytic treatment by contacting or mixing the PKC with one or more suitable enzymes under aqueous conditions, thereby degrading cell wall polysaccharides, releasing oil, smaller saccharide units and protein from the PKC.
  • the enzymes may be presented in the form of an aqueous mixture in which the enzymes are dissolved or dispersed in water, herein also referred to as an "enzyme solution” or “enzyme cocktail".
  • the water used in the enzymatic hydrolysis step is suitably purified, for example by reverse osmosis (RO).
  • RO reverse osmosis
  • one or more enzymes capable of hydrolytically cleaving one or more chemical bonds within a hemicellulose polysaccharide contained in the PKC are employed. Such enzymes may be referred to generally as hemicellulases. Hemicellulases are a diverse group of enzymes which can be categorised according to the chemical bonds they cleave: glycoside hydrolases (GH), which hydrolyse glycosidic bonds within the polysaccharide, and carbohydrate esterases (CE), which hydrolyse ester linkages within the polysaccharide.
  • GH glycoside hydrolases
  • CE carbohydrate esterases
  • GHs can be further categorised according to whether the enzyme cleaves a bond within the polysaccharide backbone (endo-glycoside) or cleaves off a mono- or di- saccharide side chain (exo-glycoside).
  • hemicellulases contemplated herein include endo-p-l,4-xylanase, exo- -l,4-xylosidase, endo- -l,4-mannanase, ⁇ - ⁇ -1,4- mannosidase, cc-arabinofuranosidase, endo-a-L-l,5-arabinase, a-glucuronidase, a- galactosidase, ⁇ -glucosidase, endo-galactanasee, acetyl xylan esterase and acetyl mannan esterase and mixtures thereof.
  • one or more other enzymes which hydrolyse other components, such as lignans, celluloses or proteins, within the PKC may also be used.
  • Some non-limiting examples thereof include lignan hydrolysing enzymes (lignases) cellulose hydrolysing enzymes (cellulases, e.g.
  • endo-l,4-p-gIucanase carboxymethyl cellulase, endo-l,4 ⁇ -D- glucanase, p-l,4-glucanase, -l,4-endoglucan hydrolase, exocellulosase and ⁇ - glucosidase), pectin hydrolysing enzymes (pectinases, e.g. polygalactouranoase), and protein hydrolysing enzymes to cleave cell wall-linked proteins (proteases). Any such one or more enzymes, crude or purified, or any mixtures thereof, are encompassed by the present disclosure.
  • Enzymes for use in the processes described herein are obtainable from microbial (e.g. fungal and bacterial) sources (e.g. Aspergillus and Tricoderma) and are commercially available.
  • Such enzymes may include a mixture of enzymes and have one or more modes of action and specificity and are commonly simply referred to as, for example, "hemicellulase”, “xylanase”, “cellulase”, “mannanase” etc.
  • some commercial "hemicellulase” typically comprises a mixture of hemicellulase enzymes, and, optionally, other enzymes, thus having multiple modes of action and specificity.
  • the skilled addressee will be able to determine suitable enzymes and combinations thereof having regard to their mode of action and specificity.
  • the hydrolysis is advantageously achieved using one or more hemicellulose degrading enzymes, e.g. hemicellulases, including mannanases.
  • hemicellulose degrading enzymes e.g. hemicellulases, including mannanases.
  • the enzymes comprise a mixture of two, three or four or more of mannanase, cellulase, pectinase, xylanase and protease.
  • Certain embodiments utilise at least one hemicellulase, such as mannanase, enzyme.
  • an enzyme mixture or cocktail is used which, in addition to mannanase, further comprises one, two, three or more of cellulase, pectinase, xylanase and protease.
  • the enzyme mixture comprises or consists of mannanase and pectinase.
  • the PKC is contacted with the enzyme or enzymes for a time and under conditions sufficient to effect at least partial degradation of the plant cell wall through hydrolysis of polysaccharide cell wall components of the PKC.
  • the skilled person will be able to determine appropriate conditions, including length of incubation period, temperature of incubation, pressure, pH conditions, quantity and combination of enzymes used and relative ratios of components.
  • the PKC and enzymes in aqueous solution may be agitated by stirring the mixture or, rotating, rocking or shaking the incubation vessel.
  • the hydrolysis can be performed under acidic, neutral or mildly basic conditions and may depend on and be adjusted as necessary by the addition of an appropriate acid or base according to the enzyme source used.
  • the hydrolysis is performed at a pH in the range of 3-8 such as pH 3-5, pH 5-7 or pH 7-8. In further examples, the hydrolysis may be performed at a pH of about 3, or 4, or, 5 or, 6, or 7, or 8. In still further embodiments, the hydrolysis is performed at a pH of about 5.
  • the hydrolysis proceeds over a period of at least about 12 hours, more advantageously, at least about 24 hours, or from about 36 or 48 hours to about 72 hours, or even up to about 168 hours.
  • longer or shorter incubation periods may be utilised depending on other factors such as the types of enzymes and amount thereof used, quantity of PKC being treated, incubation temperature etc.
  • the incubation may be carried out at any suitable temperature which allows for enzyme activity under the relevant aqueous conditions.
  • hydrolysis may be performed at a temperature in the range of about 20-80°C, such as in the range of about 25-75°C, for example about 40-60°C, or 45-55°C.
  • the incubation is performed at a temperature of about 50°C or about 60°C.
  • the ratio of PKC to aqueous enzymatic solution may depend on the nature of the enzymes and the concentration of enzymes in solution and may advantageously be anywhere from about 1 :1 to 1 :10 (w/v).
  • the ratio of PKC to aqueous enzyme solution is such that the hydrolysis mixture is sufficiently concentrated to allow hydrolysis to proceed to the desired extent under the prevailing conditions.
  • the ratio of PKC to aqueous enzymatic solution is from about 1.1 to about 1 :5, such as about 1 :2, 1 :3 or 1 :4. In further embodiments, the ratio is about 1 :3.
  • the total enzyme amount will depend on a number of factors including the nature of the enzyme(s), the amount of PKC to be treated etc, but typically may be independently in the range of about 0.1-10% (w/w) of dry PKC weight for each enzyme type, for example an amount in the range of about 0.5% (w/w) to about 3.0% (w/w), such as in the range of about 0.5% (w/w) to about 1.5% (w/w).
  • each enzyme is independently added in an amount (based on dry PKC weight) of about 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 2.0, 2.5, 3.0 or 5.0%.
  • the PKC is mixed with a cocktail of enzymes, including at least mannanase and pectinase, for example a mixture comprising mannanase, cellulase, xylanase, pectinase and optionally protease, each independently in an amount of 0.5-3.0% (w/w) on a dry PKC basis, in water, added to the PKC in a ratio of about 1:3 to 1 :5 (PKCrwater)
  • the mixture may be incubated for a period of from about 24 to 168 hours at a temperature in the range of about 40-60°C.
  • the cocktail of enzymes used is mannanase and pectinase.
  • PKC hydrolysate refers to the aqueous phase containing dissolved or dispersed products of the enzymatic hydrolysis process, including saccharides and protein, as well as palm kernel oil.
  • hydroylsed PKC also referred to herein as “kernel paste” refers to the paste-like PKC which remains after PKC has been subjected to enzymatic hydrolysis.
  • the PKC hydrolysate is separated from the hydrolysed PKC by any suitable means, including one or more of decantation, such as centrifugal decantation, centrifugation and filtration.
  • Centrifugal decantation may advantageously be carried out at any appropriate bowl speed, for example at a speed in the range of from about 2000rpm to about 5000rpm, such as about 3000rpm to about 4500rpm, at a suitable feed rate, such as from about 4, 6, 8 or lOL/min.
  • the protein may be recovered by precipitation directly from the PKC hydrolysate, in certain embodiments it is advantageous to subject the separated PKC hydrolysate to further process steps before precipitating the protein.
  • “recover” or “recovery” and variations thereof refer to the separation, isolation or purification of protein from the PKC or PKC hydrolysate.
  • a non-limiting exemplary process is depicted in Figure I.
  • the separated PKC hydrolysate may be clarified, or further separated, to separately afford an oil phase (palm kernel oil) and an aqueous protein- (and saccharide)-containing phase. Suitable methods are known in the art and may include the use of a separator or clarifier apparatus, or solvent extraction.
  • the palm oil-containing phase thus collected further increases the amount of oil obtained from the palm kernel.
  • the deactivation may be performed at any point after the hydrolysis step is completed, however, may advantageously be performed following clarification. Deactivation may be effected by heating the aqueous hydrolysate at a temperature of about 90-100°C for a time sufficient to deactivate the enzyme, for example approximately 10-15 minutes.
  • the hydrolysate is further subjected to concentration, by which is meant increasing the proportion of protein component to other components. This may be carried out by any suitable means in the art, such as membrane filtration. In some embodiments, the hydrolysate is optionally subjected to a pre-filtration step prior to further concentration.
  • the concentration of proteins is carried out by ultrafiltration, such as crossflow (tangential flow) filtration.
  • the molecular weight of proteins recovered can be controlled by selection of the appropriate sized membrane(s) to afford a "protein concentrate".
  • a protein concentrate is prepared which contains proteins having a molecular weight in the range of about 2.5kda to about 116 kda.
  • protein concentrates containing proteins of various molecular weights can be prepared, for example, having molecular weights from about 3-30 kda, or about 20-50 kda, or about 40-80 kda or about 60-100 kda or about 30-1 15 kda, such as about 20-110 kda.
  • the aqueous phase may also be subjected to further purification/separation steps. Suitable purification/separation steps may include filtration, selective adsorption on ion exchange resins or molecular sieves and chromatography.
  • the proteins may be recovered by any suitable precipitation agent/process, such as by use of an antichaotropic salt or organic solvent.
  • the protein is precipitated using antichaotropic salts.
  • Antichaotropic salts expose hydrophobic patches on proteins by removing the highly structured water layer which usually covers these patches in solution, allowing these residues to interact with each other, eventually leading to aggregation. Salts can also reduce the solubility of proteins by shielding charged groups which normally keep proteins apart in solution. When the electrostatic charge on protein molecules are shielded, the molecules can easily interact, form aggregates and eventually precipitate. The solubility of different proteins is reduced to different extents by salt addition.
  • Suitable antichaotropic salts include ammonium, sodium or potassium salts, (e.g.sulphates, phosphates or chlorides) such as ammonium sulphate, ammonium chloride, sodium sulphate and sodium chloride, potassium sulphate and potassium chloride.
  • the salt may be added as a crystalline or amorphous solid or as a further aqueous slurry or solution.
  • the salt is added to the hydrolysate,optionally clarified and/or filtered (concentrated), in a protein precipitating effective amount, which will be understood may vary with the volume and concentration of protein present in the hydrolysate.
  • Suitable concentrations of salt to add to the hydrolysate may be in the range of from about 10% (w/v) to about 90% (w/v), such as in the range of about 40-50% (w/v) to about 60-70% (w/v), for example about 65% (w/v).
  • the protein may be precipitated using an antichaotropic solvent, such as an appropriate (food grade) antichaotropic organic solvent, for example absolute ethanol.
  • the solvent may be added to the hydrolysate concentrate in any suitable amount such as from about 10% (v/v) to about 100% (v/v) or more. In some further embodiments, the solvent can be added in an amount of from about 50 or 70% (v/v) to about 80 or 90% (v/v).
  • the precipitating agent is added to the protein-containing hydrolysate in the desired precipitating effective amount and the mixture may be agitated (for example at a speed of about 50-250 rpm).for a suitable period, (for example, in the range of 1 -4, 4-8, 8-12, 12-16 or 16-24 hours), at a suitable temperature (for example, in the range of about 4-30°C, e.g. room temperature, such as about 20-25°C). In further embodiments, the mixture is agitated for about 6-14 hours, or overnight, at a temperature in the range of 4-30°C.
  • the precipitated protein is recovered by any suitable means, including filtration, decantation or centrifugation.
  • the mixture containing precipitated protein is centrifuged, for example at about 10000 rpm at a suitable temperature (e.g. about 4, 6, 8, 10, 15, 20, 25 or 30°C).
  • a suitable temperature e.g. about 4, 6, 8, 10, 15, 20, 25 or 30°C.
  • the supernatant is removed and the pellet can be re- solubilised with ethanol and centrifuged again, the process being repeated as many times as necessary to remove any excess salts
  • the recovered precipitated protein may then be freeze dried.
  • the processes described herein may be incorporated into conventional oil extraction processes and may thus be preformed on an industrial scale, such as on about 50, 100 or 200 kg or more of P C.
  • the remaining PKC e.g first or second pressed
  • the remaining PKC may be optionally subjected to one or more pre-treatment processes as described herein and transferred to a hydrolytic reactor, such as a tank or other vessel, for enzymatic hydrolysis.
  • a hydrolytic reactor such as a tank or other vessel, for enzymatic hydrolysis.
  • foreign material such as debris and large particles, may first be separated from the palm kernel.
  • the clarified hydrolysate was pre-filtered through 0.45 ⁇ Filtrafine pleated flow cartridge.
  • the resulting pre-filtered hydrolysate was then subjected to tangential flow filtration through a Millipore 10 kda membrane.
  • the retentate and permeate of the filtration were collected.
  • the collected samples were precipitate using TCA (trichloroacetic acid) methodology and analyzed in ID gel.
  • TCA trifluoroacetic acid
  • Sodium sulphate, sodium chloride and ammonium sulphate were added to 25 ml of crude aqueous hydrolysate at different concentrations of from 20% (w/v) to 50% (w/v) and stirred overnight at 100 rprn respectively. The mixture was centrifuged at 10000 rpm, at
  • Ammonium sulphate was added to 25 ml of crude aqueous hydrolysate at different concentrations of from 50% (w/v) to 90% (w/v) and stirred overnight at 100 rpm. The mixture was centrifuged at 10000 rpm, at 4 ° C for 20 minutes and the supernatant removed and the pellet re-solubilized and rewashed using 70% ethanol. The pellet (precipitated protein) was dried and weighed. The results are presented in Figure 4. The amount of precipitated protein increased with increasing concentration of ammonium sulphate used. In this particular example, the maximum effective concentration of ammonium sulphate used was 65% (w/v) which resulted 3.49 mg/g of PKC. When more than 65% (w/v) of Ammonium Sulphate was used in the sample, crystallization of the salts in the hydrolysate resulted due to the lower temperature while centrifuging the samples.
  • Protein hydrolysate (20ml) was placed into falcon tube and absolute ethanol was added at different concentrations of 10% to 100% (v/v) and the mixture was incubated at room temperature, agitating at 100 rpm, overnight. After that, the sample was centrifuged at 10000 rpm at 4 ° C. The supernatant was thawed and re-suspended and washed with 70% ethanol. The procedure was repeated three times. The pellet (precipitated protein) was dried and weighed. The results are presented in Figure 6. Use of 90% (v/v) of ethanol in the hydrolysate gave the highest amount of protein precipitated: 90.62 mg/g of PKC.
  • Protein hydrolysate (20ml) was placed into falcon tube and absolute ethanol was added at different concentrations of 10% to 100% (v/v) and the mixture was incubated at C, 8 ° C o
  • Protein hydrolysate (20ml) from a different pilot plant sample was placed into a falcon tube. Absolute ethanol at various concentration of from 10% to 100% (v/v) was added to the hydrolysate, incubated at room temperature and agitated at lOOrpm overnight. After that, sample was centrifuged at 10000 rpm at 4 ° C. The supernatant was thawed and re- suspended and washed with 70% ethanol. The procedure was repeated three times. The pellet (precipitated protein) was dried and weighed. The precipitated protein was solubilized back with RR1 buffer and loaded onto the 1D-SDS page gel . The results are presented in Figure 8. As seen in the gel image the recovered protein had a molecular weight in range of 30kda to 115kda - the band at 40kda to 60kda giving the highest protein band compared to others.
  • PKC palm kernel cake
  • GC266/Genencor 0.5% mannanase
  • pectinase Multifect Pectinase FE/ Genencor
  • the mixture was incubated at 50°C for 72 hours.
  • the mixture was separated through decantation at 2m 3 h feeding to separate the crude hydrolysate and hydrolysed PKC (decanter cake) using decanter model Aldec 30 (Alfa Laval).
  • the decanter cake was further tested for total nitrogen content in order to determine the total protein content. Three trials were run to verify the data. The results are presented in Table 1.

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Abstract

La présente invention concerne de manière générale des procédés pour le traitement de tourteaux de palmistes. Plus particulièrement, l'invention concerne des procédés d'hydrolyse catalysée par des enzymes de tourteaux de palmistes et des produits ainsi obtenus.
PCT/MY2014/000074 2013-05-21 2014-04-24 Procédés pour extraire des protéines de tourteaux de palmistes ou pour augmenter la teneur en protéines de tourteaux de palmistes WO2014189356A1 (fr)

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009025543A2 (fr) * 2007-08-20 2009-02-26 Mci Bio-Peptide Sdn. Bhd. Procédé de fabrication de polypeptides de palme
WO2009074685A1 (fr) * 2007-12-12 2009-06-18 Novozymes A/S Dégradation enzymatique de substrats de biomasse comprenant du mannane
WO2011159144A1 (fr) * 2010-06-16 2011-12-22 Malaysian Palm Oil Board Compositions contenant de l'acide shikimique obtenues à partir de matériaux à base de palmier à huile et leur procédé de fabrication

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009025543A2 (fr) * 2007-08-20 2009-02-26 Mci Bio-Peptide Sdn. Bhd. Procédé de fabrication de polypeptides de palme
WO2009074685A1 (fr) * 2007-12-12 2009-06-18 Novozymes A/S Dégradation enzymatique de substrats de biomasse comprenant du mannane
WO2011159144A1 (fr) * 2010-06-16 2011-12-22 Malaysian Palm Oil Board Compositions contenant de l'acide shikimique obtenues à partir de matériaux à base de palmier à huile et leur procédé de fabrication

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
"Production and Utilization of Protein-Enriched Palm Kernel Cake by Yeast Fermented Process for Ruminant Feeds", December 2011 (2011-12-01), XP002730165, Retrieved from the Internet <URL:http://natres.psu.ac.th/office/foreign/res/2011_Dec_ruminant.pdf> [retrieved on 20140924] *
MOHAMMAD ZAREI, AFSHIN EBRAHIMPOUR, AZIZAH ABDUL-HAMID, FAROOQ ANWAR AND NAZAMID SAARI: "Production of Defatted Palm Kernel Cake Protein Hydrolysate as a Valuable Source of Natural Antioxidants", INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, vol. 13, 29 June 2012 (2012-06-29), pages 8097 - 8111, XP002730164, ISSN: 1422-0067, DOI: 10.3390/ijms13078097 *
SCHELLER, H.V.; ULVSKOV, P., ANNU. REV. PLANT BIOL., vol. 61, 2010, pages 263 - 289

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