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WO2002088342A1 - Preparation enzymatique a valeur boulangere ajoutee et son procede de preparation - Google Patents

Preparation enzymatique a valeur boulangere ajoutee et son procede de preparation Download PDF

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Publication number
WO2002088342A1
WO2002088342A1 PCT/IN2001/000094 IN0100094W WO02088342A1 WO 2002088342 A1 WO2002088342 A1 WO 2002088342A1 IN 0100094 W IN0100094 W IN 0100094W WO 02088342 A1 WO02088342 A1 WO 02088342A1
Authority
WO
WIPO (PCT)
Prior art keywords
enzyme preparation
rhizopus
aspergillus
inoculum
matrix
Prior art date
Application number
PCT/IN2001/000094
Other languages
English (en)
Inventor
Lakshmi Ranganathan
Shiv Kumar Jayaram
Jyothi Anantha Kamath
Original Assignee
Biocon India Limited
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 Biocon India Limited filed Critical Biocon India Limited
Priority to PCT/IN2001/000094 priority Critical patent/WO2002088342A1/fr
Publication of WO2002088342A1 publication Critical patent/WO2002088342A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A21—BAKING; EDIBLE DOUGHS
    • A21D—TREATMENT, e.g. PRESERVATION, OF FLOUR OR DOUGH, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS; PRESERVATION THEREOF
    • A21D2/00—Treatment of flour or dough by adding materials thereto before or during baking
    • A21D2/08—Treatment of flour or dough by adding materials thereto before or during baking by adding organic substances
    • A21D2/24—Organic nitrogen compounds
    • A21D2/26—Proteins
    • A21D2/267—Microbial proteins
    • A—HUMAN NECESSITIES
    • A21—BAKING; EDIBLE DOUGHS
    • A21D—TREATMENT, e.g. PRESERVATION, OF FLOUR OR DOUGH, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS; PRESERVATION THEREOF
    • A21D8/00—Methods for preparing or baking dough
    • A21D8/02—Methods for preparing dough; Treating dough prior to baking
    • A21D8/04—Methods for preparing dough; Treating dough prior to baking treating dough with microorganisms or enzymes
    • A21D8/042—Methods for preparing dough; Treating dough prior to baking treating dough with microorganisms or enzymes with enzymes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/98—Preparation of granular or free-flowing enzyme compositions
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P21/00—Preparation of peptides or proteins

Definitions

  • the present invention relates to an enzyme preparation for improved baking quality and a process for preparing the same
  • the process of baking has been in existence for at least 6000 years and has been exploited ever since to improve the process.
  • the major components of baked products are polysaccharides ,proteins, fats & emulsified, enzymes, yeast and water. Of these enzymes are involved in the oxidative aging of flour, in dough development and in the formation of bread flavor & texture. They are also responsible for a number of adverse effects on the quality of the baked product. On the whole the overall effect of the indigenous enzymes of the grain, the microorganisms & of various added exogenous enzymes influence the quality of the baked product (Chemistry and physics of baking. Ed. J.M.V. Blanshard, P. J. Frazier & T. Galliard, Published by The Royal Society of Chemistry 1986).
  • Staling of baked products is a major negative influence & is defined as an increase of crumb firmness & a corresponding loss of moisture & freshness.
  • staling there is a gradual transition of the polysaccharide starch from an amorphous to a partially crystalline state. This increase in crystallinity called retrogradation is due to an inter or intra molecular association of starch molecules via hydrogen bonding .
  • the staling process is influenced by time , temperature, moisture & presence of additives in the baked product (Enzymes used in the milling industry. Ter Haseborg .E Alimenta, 1988,27, 1,2- 10).
  • Additives act by way of complexittg with the components of starch - amylose & amylopectin , thereby reducing their tendency to retrograde. Increasingly , enzymes too are being added to prevent staling. These antistaling enzymes are generally alpha & beta amylases, glucoamylases, glucose oxidase, peroxidases and lipoxygenases.
  • the antistaling enzyme should, besides aiding fresh keeping of bread should not adversely affect the other desirable parameters of a baked product like loaf volume, color, softness, resilience & crumb texture (Synergistic effect of enzymes for breadbaking . Si,J.Qi Cereal Foods World 9, 1997, 42,10,802-807).
  • Gutierrez-Correa Marcel; Tengerdy,Robert P , Biotechnol. Lett. 1998,20,1,45 - 47) and cellulases (Denada et. al., Appl Environ Microbiol, 1996,62 12,4465-70). The latter has been obtained using Trichoderma & Aspergillus strains (Mixed culture solid substrate fermentation of Trichoderma reesei with Aspergillus niger on sugarcane bagasse. Gutierrez-Correa, Marcel; Portal Leticia, Moreno Patricia, Tengerdy, Robert P, Bioresour.
  • the present invention relates to a novel process to produce an enzyme preparation with antistaling properties for improved freshkeeping of baked products.
  • the present invention provides for a A method for the manufacture of an enzyme preparation for improved baked products, which comprises of:
  • the Rhizopus sp. is Rhizopus oryzae.
  • the Aspergillus sp. is Aspergillus niger.
  • the inoculum o ⁇ Rhizopus sp. and Aspergillus sp. is mixed in the ratio ranging from 3:97 to 97:3.
  • the inoculum o ⁇ Rhizopus sp. and Aspergillus sp. is mixed in the ratio of 12.5 : 87.5.
  • the solid state nutritive matrix is selected from wheat bran, rice bran, soya grits, rice grits, millet flour, soya flour, sugar beet, bagasse or a mixture of these.
  • the mixed inoculum is optionally grown prior to mixing with sterilized solid state matrix.
  • the extraction is carried out using water or an aqueous buffer.
  • the aqueous extract is concentrated by ultra-filtration or reverse osmosis.
  • step (e) at least 10 % of inoculum is mixed with the solid state matrix.
  • step (f) the inoculate solid state matrix is incubated at 30 °C.
  • the quantity of water added Jo the extract is in the ration of 1 :6.
  • the spray dried enzyme preparation is granulated.
  • the spray dried enzyme preparation contains a preservative.
  • the preservative is selected from benzoates or sorbates.
  • the spray dried enzyme preparation contains a stabilizer.
  • the stabilizer is selected from inorganic salts, polyols, sugars or their combinations.
  • the spray drying step in 1 (i) is replaced by freeze drying or solvent precipitation.
  • the inoculum is optionally not mixed but added together to the sterilized nutritive matrix.
  • the enzyme preparation is used for baking for improved freshkeeping, whitening, softening, crumb texture or volume increase.
  • the present invention also comprises an enzyme preparation for improving baked products obtained from Rhizopus sp. and Aspergillus sp.
  • the Rhizopus sp. is Rhizopus oryzae.
  • the Aspergillus sp. is Aspergillus niger.
  • the enzyme preparation further comprises a preservative, preferably selected from benzoates or sorbates.
  • the enzyme preparation further comprises a stabilizer, preferably selected from inorganic salts, polyols, sugars or their combinations .
  • the present invention has the following advantages over the other reported methods: (i) The enzyme preparation obtained is from a non-genetically modified
  • spore suspension o ⁇ Rhizopus oryzae made by adding 3 mL of sterile water to a culture slant, is added to a 250 mL Erlenmeyer flask containing 35 mL of medium with following composition:
  • the flasks are incubated at 30 deg C on a rotary shaker (200-rpm) for 48 hours.
  • the flasks are incubated at 30 deg C on a rotary shaker (200-rpm) for 48 hours.
  • the inoculum developed from Example 1 is mixed the inoculum of Example 2 in the ratio 12.5:87.5. This mixed inoculum is used in the subsequent stages.
  • Moist wheat bran is autoclaved for 90minutes in a rotating cooker.
  • the mixed inoculum from example 3 is mixed with the sterilized wheat bran from example 4. A 10 % inoculum is used. The inoculum is mixed at 30 deg. C. After a proper mixing, the inoculated wheat bran is transferred and layered into the pre- sterilized trays.
  • Example 6 Incubation: Intubation of the layered trays from example 5 is done at 30 deg.C. for 5 days. At the end of 5 days, the fermented solid substrate is harvested.
  • Extraction is done using water. Quantity of water added to the extract is in a ratio of 1:6. Soaking of the harvested solid substrate is done for 6 hours.
  • Microfiltartion is done to the extract obtained from example 7 to obtain a microorganism free product. This is done using a 0.2 micron MF membrane.
  • the resultant liquid obtained from example 9 is concentrated to the desired activity by Ultrafiltration done at 15 deg.C. Ultrafiltration is done using UF membranes.
  • Example 11 Enzyme stabilization J ⁇ e Ultrafiltered enzyme obtained from example 10 is stabilized by the addition ofNaCl and KCl.
  • the Ultrafiltered enzyme obtained from example 10 is stabilized by the addition of gylcerol and sorbitol.
  • the UFC (ultrafiltered concentrate) as obtained from example 1 is spray dried to get the required enzyme preparation.
  • the UFC (ultrafiltered concentrate) as obtained from Example 11 is spray dried as in Example 13, but with the addition of 0.1 % potassium sorbate as a preservative before spray drying.
  • the UFC (ultrafiltered concentrate) as obtained from Example 11 is freeze dried.
  • Example 16 Baking trials The enzyme preparation obtained from example 15 was used in a baking trial. This freeze dried enzyme was dosed at 113 ppm in a bread baking trial & was evaluated for fresh keeping against a trial which did not have the enzyme.
  • the bread made with the freeze dried enzyme preparation appeared softer than that of the control even after 14 days.

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  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • General Health & Medical Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Food Science & Technology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Botany (AREA)
  • Mycology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Virology (AREA)
  • Enzymes And Modification Thereof (AREA)

Abstract

La présente invention concerne un procédé de préparation amélioré d'un mélange enzymatique destiné à améliorer les caractéristiques de conservation de produits cuits au four. Cette préparation enzymatique est produite par coculture de deux espèces fongiques de Rhizopus & Aspergillus par fermentation en milieu solide dans des conditions de fermentation optimales. Le produit extrait subit un traitement ultérieur permettant d'obtenir une préparation enzymatique sous forme de poudre compacte. La présente invention concerne également une préparation enzymatique obtenue à partir de Rhizopus sp. et Aspergillus sp..
PCT/IN2001/000094 2001-04-30 2001-04-30 Preparation enzymatique a valeur boulangere ajoutee et son procede de preparation WO2002088342A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
PCT/IN2001/000094 WO2002088342A1 (fr) 2001-04-30 2001-04-30 Preparation enzymatique a valeur boulangere ajoutee et son procede de preparation

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/IN2001/000094 WO2002088342A1 (fr) 2001-04-30 2001-04-30 Preparation enzymatique a valeur boulangere ajoutee et son procede de preparation

Publications (1)

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WO2002088342A1 true WO2002088342A1 (fr) 2002-11-07

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004113490A2 (fr) * 2003-06-24 2004-12-29 HöFer Bioreact GmbH Culture selective et dirigee, en phase solide, de populations microbiennes mixtes stables pour la preparation en continu de melanges enzymes - metabolites determines
JP2012139150A (ja) * 2010-12-28 2012-07-26 Kitakyushu Foundation For The Advancement Of Industry Science & Technology アスペルギルス属菌及びリゾープス属菌の混合培養系を用いたアミラーゼの生産方法
US8663720B2 (en) * 2004-03-12 2014-03-04 Nippon Supplement, Inc. Process for preparing fermented tea and composition

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4308284A (en) * 1979-04-27 1981-12-29 Kikkoman Shoyu Co. Ltd. Process for producing koji for fermented food products
JPS6170980A (ja) * 1984-06-30 1986-04-11 Niigataken 変り麹の製造法
EP0585988A1 (fr) * 1992-07-27 1994-03-09 Gist-Brocades N.V. Produit enzymatique et méthode d'amélioration de la qualité du pain
EP0913092A2 (fr) * 1997-10-31 1999-05-06 Amano Pharmaceutical Co., Ltd. Composition de pâte et sa préparation
WO1999028366A1 (fr) * 1997-12-04 1999-06-10 Board Of Regents Of University Of Nebraska Polyesters degradables
EP1022329A1 (fr) * 1999-01-25 2000-07-26 GIE Agro Industrie Produit multienzymatique à activités glucoamylasique, protéolytique et xylanasique et procédé pour sa production par fermentation à l'état solide de son de blé avec Aspergillus niger

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4308284A (en) * 1979-04-27 1981-12-29 Kikkoman Shoyu Co. Ltd. Process for producing koji for fermented food products
JPS6170980A (ja) * 1984-06-30 1986-04-11 Niigataken 変り麹の製造法
EP0585988A1 (fr) * 1992-07-27 1994-03-09 Gist-Brocades N.V. Produit enzymatique et méthode d'amélioration de la qualité du pain
EP0913092A2 (fr) * 1997-10-31 1999-05-06 Amano Pharmaceutical Co., Ltd. Composition de pâte et sa préparation
WO1999028366A1 (fr) * 1997-12-04 1999-06-10 Board Of Regents Of University Of Nebraska Polyesters degradables
EP1022329A1 (fr) * 1999-01-25 2000-07-26 GIE Agro Industrie Produit multienzymatique à activités glucoamylasique, protéolytique et xylanasique et procédé pour sa production par fermentation à l'état solide de son de blé avec Aspergillus niger

Non-Patent Citations (9)

* Cited by examiner, † Cited by third party
Title
BIGELIS R: "Handbook of applied mycology, Volume 3: Foods and feeds (eds Arora D K et al.), Chapter 14: FUNGAL ENZYMES IN FOOD PROCESSING (pp 445-498)", 1991, MARCEL DEKKER, INC., NEW YORK, USA; ISBN 0-8247-8491-X., XP001051671 *
BOWLES L K: "Amylolytic enzymes. In: Baked goods freshness. Technology, evaluation, and inhibition of staling (pp 105-129).", 1996, MARCEL DEKKER, INC., NEW YORK, USA, XP002186664 *
DATABASE WPI Section Ch Week 198621, Derwent World Patents Index; Class D16, AN 1986-134353, XP002185477 *
ENZYME BUSINESS NOVO NORDISK A/S DENMARK: "Raising the standard of baking enzymes.", WORLD OF INGREDIENTS, November 1995 (1995-11-01), pages 50, XP001051360 *
GRUNDY J G: "Preservatives. In: Baked goods freshness. Technology, evaluation, and inhibition of staling (pp 189-203).", 1996, MARCEL DEKKER, INC., NEW YORK, USA, XP002186666 *
KIM H-S ET AL.: "Enzymological characteristics and identification of useful fungi isolated from traditional Korean Nuruk.", KOREAN JOURNAL OF APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, vol. 26, no. 5, 1998, pages 456 - 464, XP001051554 *
OOIJKAAS L P ET AL.: "Defined media and inert supports: their potential as solid-state fermentation production systems", TRENDS IN BIOTECHNOLOGY, vol. 18, no. 8, 1 August 2000 (2000-08-01), pages 356 - 360, XP004213293, ISSN: 0167-7799 *
POTUS J: "L'intérêt des enzymes en panification. Role of enzymes in breadmaking", COMPTES RENDUS DE L'ACADEMIE D'AGRICULTURE DE FRANCE, vol. 81, no. 2, 1995, pages 27 - 36, XP000946072, ISSN: 0989-6988 *
VAN EIJK J H ET AL.: "Nonamylolytic enzymes. In: Baked goods freshness. Technology, evaluation, and inhibition of staling (pp 131-150)", 1996, MARCEL DEKKER, INC., NEW YORK, USA, XP002186665 *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004113490A2 (fr) * 2003-06-24 2004-12-29 HöFer Bioreact GmbH Culture selective et dirigee, en phase solide, de populations microbiennes mixtes stables pour la preparation en continu de melanges enzymes - metabolites determines
WO2004113490A3 (fr) * 2003-06-24 2005-07-28 Hoefer Bioreact Gmbh Culture selective et dirigee, en phase solide, de populations microbiennes mixtes stables pour la preparation en continu de melanges enzymes - metabolites determines
US9127299B2 (en) 2003-06-24 2015-09-08 Senzyme Gmbh Directed selective solid-phase culturing of stable microbial mixed populations for the continuous preparation of defined enzyme and metabolite mixtures
US8663720B2 (en) * 2004-03-12 2014-03-04 Nippon Supplement, Inc. Process for preparing fermented tea and composition
JP2012139150A (ja) * 2010-12-28 2012-07-26 Kitakyushu Foundation For The Advancement Of Industry Science & Technology アスペルギルス属菌及びリゾープス属菌の混合培養系を用いたアミラーゼの生産方法

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