WO2002069732A1 - Procede de preparation d'hydrolysat de proteine a partir de farine de soja - Google Patents

Procede de preparation d'hydrolysat de proteine a partir de farine de soja Download PDF

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Publication number
WO2002069732A1
WO2002069732A1 PCT/IN2001/000031 IN0100031W WO02069732A1 WO 2002069732 A1 WO2002069732 A1 WO 2002069732A1 IN 0100031 W IN0100031 W IN 0100031W WO 02069732 A1 WO02069732 A1 WO 02069732A1
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WO
WIPO (PCT)
Prior art keywords
protein hydrolysate
hydrolysis
protein
soy flour
hydrolysate
Prior art date
Application number
PCT/IN2001/000031
Other languages
English (en)
Inventor
Appu Rao Gopala Rao Appu Rao
Karadka Govindaraju
Ramaswamy Harendranath
Johny Joseph
Vishweshwariah Prakash
Cheruppanpullil Radha
Mysore Cheluvaraya Shamanthaka Sastry
Sridevi Annapurna Singh
Original Assignee
Council Of Scientific And Industrial Research
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Council Of Scientific And Industrial Research filed Critical Council Of Scientific And Industrial Research
Priority to CN01823124.1A priority Critical patent/CN1234288C/zh
Priority to PCT/IN2001/000031 priority patent/WO2002069732A1/fr
Publication of WO2002069732A1 publication Critical patent/WO2002069732A1/fr
Priority to ZA200306957A priority patent/ZA200306957B/en

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
    • C12Y304/22—Cysteine endopeptidases (3.4.22)
    • C12Y304/22002—Papain (3.4.22.2)
    • 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
    • A23J3/16—Vegetable proteins from soybean
    • 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
    • A23J3/346—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of vegetable proteins

Definitions

  • the present invention relates to a process for the preparation of protein hydrolysate from soy flour using fungal protease. Particularly, the present invention relates to a process for the preparation of protein hydrolysate from defatted soy flour using fungal protease obtained from Aspergillus sp.
  • soybean A small portion of total soybean produced also finds use for different edible grade flours, protein isolate and texturized products and popularity of these products are greatly picking up globally. Soybean is an excellent source of protein, which contains about 40% protein. New manufacturing techniques for high quality soybean foods have been developed by lowering or destroying of the anti-nutritional factors like trypsin inhibitors.
  • United States Patent No. 5180597 claimed a process for hydrolyzed vegetable protein with enhanced flavor, which contains no detectable level of monochlorodihydroxypropanol, is described.
  • wheat gluten is hydrolyzed using Prozyme 6 (a fungal protease) at a temperature of 40-50°C, pH 6.5-7.0, enzyme concentration of 0.1-2.0% of substrate for a time period of 4h.
  • the hydrolyzed protein is treated with gaseous HC1 for deamidation before the addition of acid for inactivating the enzyme.
  • the drawback in such hydrolysis is that it is likely to lead to racemisation of amino acids and the addition of acid increases the salt content in the product.
  • Alcalase is the enzyme used.
  • the drawback of the process is that it is a multi step process and due to partial defatting of soy flour, left over oil comes in contact with protein phase which could lead to off-flavors.
  • Enzyme inactivation is done by addition of acid, which is likely to lead to increased salt content in the product.
  • the soy seeds are comminuted and heated at 90-140°C.
  • Protease fungal and bacterial
  • the fiber is separated and slurry has two phases- oil and aqueous phase.
  • Aqueous phase is brought to pH 4.5 to precipitate the protein, which is then concentrated.
  • the starting material is not defatted and hence the residual oil could come in contact with the aqueous phase, which could lead to off-flavors.
  • the main object of the present invention is to provide a process for the preparation of protein hydrolysate from soy flour using microbial protease.
  • Another object of the present invention is to provide a process for the preparation of protein hydrolysate from defatted soy flour using Aspergillus sp. .
  • Still another object of the present invention is to provide a process for preparing protein hydrolysate by double enzyme hydrolysis using proteolytic enzymes.
  • Yet another object of the present invention is to decrease the bitterness in the hydrolysate to the extent that the threshold perception of bitterness is greater than 2g %.
  • One more object of the present invention is to provide a process for the preparation of protein hydrolysates with a low mineral content
  • One another object of the present invention is to provide a process for the preparation of protein hydrolysate in higher yield and with a specified degree of hydrolysis from the raw material taken.
  • the present invention provides a process for the preparation of protein hydrolysate from soy flour, which comprises preparing aqueous slurry of defatted soy flour having 6-12% w/v of solid content, hydrolyzing the said slurry using fungal protease at pH 7-8 and temperature 43 ⁇ 5°C to get 20-40% DH, further hydrolyzing using papain at temperature 53 ⁇ 5°C under stirring till completion of hydrolysis to 30 - 45% DH, inactivating residual enzyme in a known manner, separating the solids and drying the clarified supernatant thus obtained to get protein hydrolysate.
  • the present invention provides a process for the preparation of protein hydrolysate from soy flour using fungal protease, said process comprising: preparing aqueous slurry of defatted soy flour having 6-12% w/v of solid content; hydrolyzing the said slurry using fungal protease at pH 7-8 and temperature 43+ 5°C to get 20-40% degree of hydrolysis (DH); further hydrolyzing using papain at temperature 53 ⁇ 5°C under stirring till 30 - 45% DH is obtained; inactivating residual enzyme in a known manner; separating the solids and drying the clarified supernatant thus obtained to get protein hydrolysate.
  • DH degree of hydrolysis
  • the solid content in the slurry ranges from 8 -
  • the fungal protease is obtained from Aspergillus sp.
  • t e Aspergillus is selected from the group comprising of A. flaws, A. japanicus, A. niger and A. awamori.
  • protein hydrolysate is obtained by double enzyme hydrolysis.
  • protein hydrolysate is obtained by hydrolyzing the slurry with proteolytic enzyme.
  • the fungal protease used ranges from
  • protease hydrolysis is carried out at a pH of 7.2 to 7.6.
  • the amount of papain used ranges from 0.4 to 0.5% w/w of the soy flour.
  • the hydrolysate produced has decreased bitterness.
  • the threshold perception of bitterness is greater than 2g %.
  • the protein hydrolysate produced has low mineral content.
  • high yield of protein hydrolysate with 35 to 45% degree of hydrolysis is obtained from the raw material taken.
  • soy protein hydrolysate obtained has creamy color and a yield of 60-67.0% (on protein basis).
  • soy protein hydrolysate has 9.4% moisture, 10.5-11.0% nitrogen and 35-45% degree of hydrolysis.
  • the soy protein hydrolysate obtained has 20-23 trypsin inhibitor units/mg activity, 95 to 98% Nitrogen Solubility Index, 0.6 to 1.0% of salt content and 2 to 2.2 % bitterness recognition threshold.
  • lipoxygenase and urease activities of the protein hydrolysate were not detectable.
  • the amino acid composition of the protein hydrolysate is similar to the amino acid makeup of starting material.
  • the protein hydrolysate is creamy in color.
  • the present invention also provides a protein hydrolysate having 60 to 67% protein hydrolysate on protein basis.
  • the protein hydrolysate has 9.4% moisture, 10.5 to 11% nitrogen, and 35 to 45 % degree of hydrolysis.
  • the protein hydrolysate has 20 to 23 trypsin inhibitor units/mg activity, 95 to 98% Nitrogen Solubility Index, 0.6 to 1.0% salt content and 2 to 2.2% bitterness recognition threshold.
  • the protein hydrolysate has undetectable lipoxtgenase and urease activities.
  • the protein hydrolysate has similar acid makeup as that of the starting material
  • Soybean flour is derived from clean round beans. The cleaned beans are passed through a cracking process; the bean fragments are graded on sieves and aspirate system. The cleaned, cracked meat is passed through a conditioner cooker and flaked. This is subjected to solvent extraction process. The extracted flakes were desolventized and ground to 100 mesh.
  • the specification for soy flour consists of (a) Moisture % by mass, max. 9% (b) Protein on dry basis % by mass, min. 48%, (c) total ash on dry basis, % by nmass max. 7.2%, (d) acid insoluble ash on dry basis % by mass max. 0.4 %, (e) fat on dry basis % by mass,
  • DH Degree of Hydrolysis
  • Trinitrobenzenesulphonic acid (TNBS) procedure is an accurate, reproducible and generally applicable procedure for determining the degree of hydrolysis of food protein hydrolysates.
  • the protein hydrolysate is dissolved/dispersed in hot 1% sodium dodecyl sulphate to a concentration of 0.25 - 2.5 x 10 "3 amino equivalents/L.
  • a sample solution (0.25 ml) is mixed with 2 ml of 0.2125 M sodium phosphate buffer (pH 8.2) and 2 ml of
  • soy flour is dispersed in 10 L of water and the pH of the dispersion was adjusted .to 7.6. It was kept stirring for 15 minutes with mechanical stirrer and then temperature raised to 45°C. At this stage, 5 g of fungal protease is added and stirring continued for 2 hours. At the end of 2 hours, the slurry temperature was raised to 55°C and the second enzyme papain (5g) was added and kept stirring for 1.5 hours. After hydrolysis, the hydrolysate was boiled for 15 minutes for enzyme inactivation and centrifuged in the basket centrifuge. The clear solution was spray dried. The degree of hydrolysis was found to be 38 % and the yield was 70% on protein basis.
  • the particle size of the soy flour, ratio of enzyme to substrate, temperature, pH and time interval controls the end of enzymatic hydrolysis resulting into minimizing bitterness of the hydrolysate.
  • the soy protein hydrolysate obtained has creamy color and an yield of 60-67.0% (on protein basis).
  • the product has 9.4% moisture, 10.5-11.0% nitrogen and 35-45% degree of hydrolysis (TNBS procedure).
  • the soy protein hydrolysate obtained has 20-23 trypsin inhibitor units/mg activity (TIU), 95-98% Nitrogen Solubility Index, 0.6-1.0% of salt content (measured as CI " ions) and 2- 2.2% bitterness recognition threshold.
  • the lipoxygenase and urease activities were not detectable.
  • the amino acid composition of the soy protein hydrolysate obtained was similar to the amino acid make up of starting raw material thereby retaining the nutritional value.
  • the protein hydrolysate is less bitter compared to protein hydrolysate obtained from casein and is less hygroscopic in nature.
  • the main advantages of this invention are -
  • the process provides protein hydrolysate with specified degree of DH and 10.5 to 11.0% nitrogen content in a powder form.
  • the product attains a property of becoming a good additive without imparting any undesirable odd flavour for the finished product.
  • the process yields a quality hydrolysate which has a solubility independent of pH making it a suitable additive either in acid pH or alkaline pH.
  • the Nitrogen recovery from the soy flour is 95-98%) which is higher compared to any present method of commercial production.
  • the yield of protein hydrolyste is 65-70%> of the soy bean protein.
  • the time of hydrolysis is 2 l A to 4 h. which is short having advantage both in input cost and energy.
  • the combination of the two enzymes is such that salt concentration will be minimal at 0.6 to 1%.
  • the enzyme employed is a food grade commercially available acceptable plant enzyme/fungal enzyme with broad specificity.
  • the protein hydrolysate so obtained contained acceptable levels of trypsin inhibitor activity, free from lipoxygenase activity and urease activity and NSI is 98%.

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Organic Chemistry (AREA)
  • Nutrition Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Genetics & Genomics (AREA)
  • General Chemical & Material Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Beans For Foods Or Fodder (AREA)

Abstract

L'invention concerne un procédé de préparation d'hydrolysat de protéine à partir de farine de soja. Ce procédé consiste: à préparer une pâte aqueuse de soja déshuilé présentant de 6 à 12 % M/V de matière solide; à hydrolyser ladite pâte au moyen d'une protéase fongique, à un pH de 7-8 et à une température de 43 ? 5 °C, pour obtenir un degré d'hydrolyse (DH) compris entre 20 et 40 %; à continuer l'hydrolyse au moyen d'une papaïne, à une température de 53 ? 5 °C, en brassage, jusqu'à ce que l'hydrolyse atteigne un DH compris entre 30 et 45 %; à inactiver l'enzyme résiduelle selon un procédé connu; à séparer les solides et à sécher le supernageant clarifié ainsi obtenu, de sorte à obtenir l'hydrolysat de protéine.
PCT/IN2001/000031 2001-03-05 2001-03-05 Procede de preparation d'hydrolysat de proteine a partir de farine de soja WO2002069732A1 (fr)

Priority Applications (3)

Application Number Priority Date Filing Date Title
CN01823124.1A CN1234288C (zh) 2001-03-05 2001-03-05 从大豆粉中制备蛋白质水解产物的方法
PCT/IN2001/000031 WO2002069732A1 (fr) 2001-03-05 2001-03-05 Procede de preparation d'hydrolysat de proteine a partir de farine de soja
ZA200306957A ZA200306957B (en) 2001-03-05 2003-09-05 Process for preparation of protein hydrolysate from soy flour.

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/IN2001/000031 WO2002069732A1 (fr) 2001-03-05 2001-03-05 Procede de preparation d'hydrolysat de proteine a partir de farine de soja

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WO2002069732A1 true WO2002069732A1 (fr) 2002-09-12

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CN (1) CN1234288C (fr)
WO (1) WO2002069732A1 (fr)
ZA (1) ZA200306957B (fr)

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2011006916A3 (fr) * 2009-07-17 2011-03-17 Novozymes A/S Produit a base de soja modifie par traitement enzymatique
US8900348B2 (en) 2009-07-02 2014-12-02 Johnson Matthey Public Limited Company Adsorption of volatile organic compounds derived from organic matter
US9186649B2 (en) 2005-11-01 2015-11-17 Anglo Platinum Marketing Limited Adsorption of volatile organic compounds derived from organic matter
RU2612153C1 (ru) * 2016-06-07 2017-03-02 Василий Васильевич Пономарев Способ интенсификации массообменных процессов
RU2612155C1 (ru) * 2015-11-26 2017-03-02 Василий Васильевич Пономарев Способ получения белковых гидролизатов
FR3076982A1 (fr) * 2018-01-23 2019-07-26 Laboratoires Gilbert Hydrolysat proteique de fabacees comme source proteique hypoallergenique dans des compositions alimentaires
CN112940842A (zh) * 2021-03-19 2021-06-11 东北农业大学 一种利用两步酶解制备低苦味豆粉和高品质豆油的方法

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103194517B (zh) * 2013-04-11 2016-04-06 中国科学院过程工程研究所 一种鹰嘴豆短肽混合物及其制备方法
CA3006448A1 (fr) * 2015-12-18 2017-06-22 Nestec S.A. Compositions thermosterilisees a haute teneur en proteines contenant une proteine hydrolysee issue d'un processus continu avec au moins une endopeptidase

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4324805A (en) * 1979-07-11 1982-04-13 Novo Industri A/S Method of producing soy protein hydrolysate from fat-containing soy material, and soy protein hydrolysate
EP0087247A2 (fr) * 1982-02-22 1983-08-31 Stauffer Chemical Company Procédé de préparation d'hydrolysats de protéines
EP0325986A2 (fr) * 1988-01-28 1989-08-02 Miles Inc. Hydrolyse enzymatique des protéines
WO1994025580A1 (fr) * 1993-04-26 1994-11-10 Novo Nordisk A/S Procede d'hydrolyse de proteines
WO1998018342A1 (fr) * 1996-10-30 1998-05-07 Novo Nordisk A/S Procede de preparation d'un agent aromatisant alimentaire
US6221423B1 (en) * 1998-04-13 2001-04-24 Protein Technologies Int'l Inc. Short-chained peptide material

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4324805A (en) * 1979-07-11 1982-04-13 Novo Industri A/S Method of producing soy protein hydrolysate from fat-containing soy material, and soy protein hydrolysate
EP0087247A2 (fr) * 1982-02-22 1983-08-31 Stauffer Chemical Company Procédé de préparation d'hydrolysats de protéines
EP0325986A2 (fr) * 1988-01-28 1989-08-02 Miles Inc. Hydrolyse enzymatique des protéines
WO1994025580A1 (fr) * 1993-04-26 1994-11-10 Novo Nordisk A/S Procede d'hydrolyse de proteines
WO1998018342A1 (fr) * 1996-10-30 1998-05-07 Novo Nordisk A/S Procede de preparation d'un agent aromatisant alimentaire
US6221423B1 (en) * 1998-04-13 2001-04-24 Protein Technologies Int'l Inc. Short-chained peptide material

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9186649B2 (en) 2005-11-01 2015-11-17 Anglo Platinum Marketing Limited Adsorption of volatile organic compounds derived from organic matter
US8900348B2 (en) 2009-07-02 2014-12-02 Johnson Matthey Public Limited Company Adsorption of volatile organic compounds derived from organic matter
WO2011006916A3 (fr) * 2009-07-17 2011-03-17 Novozymes A/S Produit a base de soja modifie par traitement enzymatique
AU2010272591B2 (en) * 2009-07-17 2015-02-12 Novozymes A/S Enzyme-modified soybean product
US9089154B2 (en) 2009-07-17 2015-07-28 Novozymes A/S Enzyme-modified soybean product
RU2612155C1 (ru) * 2015-11-26 2017-03-02 Василий Васильевич Пономарев Способ получения белковых гидролизатов
RU2612153C1 (ru) * 2016-06-07 2017-03-02 Василий Васильевич Пономарев Способ интенсификации массообменных процессов
FR3076982A1 (fr) * 2018-01-23 2019-07-26 Laboratoires Gilbert Hydrolysat proteique de fabacees comme source proteique hypoallergenique dans des compositions alimentaires
WO2019145351A1 (fr) * 2018-01-23 2019-08-01 Laboratoires Gilbert Hydrolysat proteique de fabacees comme source proteique hypoallergenique dans des compositions alimentaires
CN112940842A (zh) * 2021-03-19 2021-06-11 东北农业大学 一种利用两步酶解制备低苦味豆粉和高品质豆油的方法

Also Published As

Publication number Publication date
CN1234288C (zh) 2006-01-04
ZA200306957B (en) 2004-07-28
CN1494382A (zh) 2004-05-05

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