WO2002088342A1 - An enzyme preparation for improved baking quality and a process for preparing the same - Google Patents
An enzyme preparation for improved baking quality and a process for preparing the same Download PDFInfo
- 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
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.
Landscapes
- 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
The present invention provides an improved method for the preparation of an enzyme mixture for improved freshkeeping of baked products. The enzyme preparation is produced by the co culture of two fungal species of Rhizopus & Aspergillus by solid substrate fermentation under optimal fermentation parameters. The product is extracted and further processed to get an enzyme preparation as solid powder. The present invention also includes an enzyme preparation obtained from obtained from Rhizopus sp. and Aspergillus sp..
Description
AN ENZYME PREPARATION FOR IMPROVED BAKING QUALITY AND A PROCESS FOR PREPARING THE SAME
The present invention relates to an enzyme preparation for improved baking quality and a process for preparing the same
BACKGROUND
The process of baking has been in existence for at least 6000 years and has been exploited ever since to improve the process. TThe 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. During 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.
For a successful antistaling enzyme it is essential that they can act on raw starch rather than gelatinised starch & have an intermediate heat stability i.e. more stable than the conventional heat labile fungal amylases but less stable than the conventional heat stable bacterial amylases. These enzymes would thus provide an antistaling effect without adversely affecting the quality of the baked product , whereby they hydrolyse starch during the dough development process, but are completely heat inactivated before the baking process is completed. They thus prevent an excess of the breakdown products of starch - the dextrins which lead to an increased stickiness & gumminess of the bread (Enzymes for the baking industry . Van Oor M.G, Hamer, R.J Aliment. ,Equipos Tecnol,1993, 12,5,115 -18). 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).
Co culture of different strains of bacteria - bacteria (Deanda et. al., Appl Environ Microbiol, 1996,62,12,4465-70), yeast -bacteria (Kim et. al., Biotechnol Lett, 1996, 18,9, 1031 -34), yeast -yeast , yeast / bacteria - fungi (Padmaja et. al. J. Sci Food Agric, 1993, 63,4,473-81; Marakis S.G Biotechnol Lett, 1992,14,11,1075-80) & fungi -fungi (Benzuela Elegado, Francisco, Fujio,Yusaku J. Gen. Appl.
Microbiol.1993,39,6, 541-6; Gutierrez-Correa, Marcel; Tengerdy,Robert P , Biotechnol. Lett. 1997,19,7,665-667; Cellulase production by mixed fungi in solid substrate fermentation of bagasse (Duenas R et. Al. World T. Microbiol. Biotechnol ,1995 ,11,3,333-337) have been reported in literature. . These associations have been used to produce many specialty products like acetic acid, alcohol butanol & enzymes like xylanase (Xylanase production by fungal mixed culture solid substrate fermentation on sugarcane bagasse. 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. Technol , 1998, Volume date 1999, 68,2,173-178; Production of cellulase on sugarcane bagasse by fungal mixed culture solid substrate fermentation. Gutierrez-Correa, Marcel; Tengerdy ,Robert P , Biotechnol. Lett. 1997,19,7,665-667; Formation of cellulases by co - culturing of Trichoderma reesei and Aspergillus niger on cellulosic waste. Madamwar,D ,Patel,S, World J. Mierobiol.Biotechnol 1992, 8,2,183-6).
DESCRIPTION OF THE INVENTION
The present invention relates to a novel process to produce an enzyme preparation with antistaling properties for improved freshkeeping of baked products.
Accordingly, the present invention provides for a A method for the manufacture of an enzyme preparation for improved baked products, which comprises of:
a. preparing an inoculum of Rhizopus sp.,
b. preparing an inoculum oϊ Aspergillus sp., c. mixing the grown inoculum of Rhizopus sp. and Aspergillus sp., ά. sterilizing a solid state nutritive matrix, e. mixing the said sterilized solid state nutritive matrix with the mixture of inoculum consisting of Rhizopus sp. and Aspergillus sp., f. incubating the said inoculated solid state nutritive matrix for 4 - 7 days at 25 - 35°C, g. extracting the fermented matrix followed by filtration, h. concentrating the aqueous extract, i. spray drying of the concentrated aqueous extract to get the enzyme preparation.
The Rhizopus sp. is Rhizopus oryzae. The Aspergillus sp. is Aspergillus niger. The inoculum oϊ Rhizopus sp. and Aspergillus sp. is mixed in the ratio ranging from 3:97 to 97:3. The inoculum oϊ 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. In step (e) at least 10 % of inoculum is mixed with the solid state matrix. In 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
(GMO) origin, (ii) The process is environment friendly, less cumbersome and economical for large-scale industrial applications, (iii) A broader range of enzyme activities that aid freshkeeping is obtained , which is not obtained with a single culture alone, (iv) This combination of desired activities for freshkeeping is generated in situ thereby having an advantage over blended enzyme preparations, (v) The enzyme preparation is capable of acting on ra starch , rather than gelatinized starch.
The invention will now be described with reference to the following examples:
EXAMPLE 1
Preparation of inoculum of Rhizopus sp:
About 100 μL spore suspension oϊ 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.
EXAMPLE 2
Preparation of inoculum of Aspergillus sp:
About 100 μL spore suspension oϊ Aspergillus niger, 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.
EXAMPLE 3
Mixing of the inoculum
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.
Example 4
Sterilization of wheat bran:
Commercial wheat bran is used as the solid substrate. Moist wheat bran is autoclaved for 90minutes in a rotating cooker.
Example 5 Inoculation:
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.
Example 7 Extraction:
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.
Example 8 Extraction:
The extraction is carried out in the same way as example 7 but with the addition of 0.2% sodium benzoate with the water.
Example 9 Microfiltration:
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.
(Xxample 10 Ultrafiltration:
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:
Jπe Ultrafiltered enzyme obtained from example 10 is stabilized by the addition ofNaCl and KCl.
Example 12 Enzyme stabilization:
The Ultrafiltered enzyme obtained from example 10 is stabilized by the addition of gylcerol and sorbitol.
Example 13 Spray drying:
The UFC (ultrafiltered concentrate) as obtained from example 1 is spray dried to get the required enzyme preparation.
Example 14 Spray drying:
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.
Example 15 Freeze 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 results of the baking trials of the test sample (containing the enzyme preparation), were compared with the control (where the enzyme preparation was absent) and the quality of the bread evaluated. The following observations were made:
Note: + denotes poor
++ denotes average +-H- denotes good - denotes absent
n
Claims
1. A method for the manufacture of an enzyme preparation for improved baked products, which comprises of:
j . preparing an inoculum of Rhizopus sp. , k. preparing an inoculum oϊ Aspergillus sp.,
1. mixing the grown inoculum oϊ Rhizopus sp. and Aspergillus sp. , m. sterilizing a solid state nutritive matrix, n. mixing the said sterilized solid state nutritive matrix with the mixture of inoculum consisting oϊ Rhizopus sp. and Aspergillus sp., o. incubating the said inoculated solid state nutritive matrix for 4 - 7 days at 25 - 35 p. extracting the fermented matrix followed by filtration, q. concentrating the aqueous extract, r. spray drying of the concentrated aqueous extract to get the enzyme preparation.
2. The method as claimed in claim 1 wherein the Rhizopus sp. is Rhizopus oryzae.
3. The method as claimed in claim 1 wherein the Aspergillus sp. is Aspergillus niger.
4. The method as claimed in claim 1 wherein the inoculum oϊ Rhizopus sp. and Aspergillus sp. is mixed in the ratio ranging from 3:97 to 97:3.
5. The method as claimed in claim 4 wherein the inoculum oϊ Rhizopus sp. and Aspergillus sp. is mixed in the ratio of 12.5 : 87.5.
6. The method as claimed in claim 1 wherein 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.
7. The method as claimed in claim 1 wherein the mixed inoculum is optionally grown prior to mixing with sterilized solid state matrix.
8. The method as claimed in claim 1 wherein the extraction is carried out using water or an aqueous buffer.
9. The method as claimed in claim 1 wherein the aqueous extract is concentrated by ultra-filtration or reverse osmosis.
10. The method as claimed in claim 1 wherein in step (e) at least 10 % of inoculum is mixed with the solid state matrix.
11. The method as claimed in claim 1 wherein in step (ϊ), the inoculate solid state matrix is incubated at 30 °C.
12. The method as claimed in claim 8 wherein quantity of water added to the extract is in the ration of 1 :6.
13. The method as claimed in claim i wherein the spray dried enzyme preparation is granulated.
14. The method as claimed in claim 1 or 13 wherein the spray dried enzyme preparation contains a preservative.
15. The method as claimed in claim 14 wherein the preservative is selected from benzoates or sorbates.
16 The method as claimed in any one of the preceding claims wherein the spray dried enzyme preparation contains a stabilizer.
17. The method as claimed in claim 16 wherein the stabilizer is selected from inorganic salts, polyols, sugars or their combinations .
18. The method as claimed in claim 1 to 17 wherein the spray drying step in 1 (i) is replaced by freeze drying or solvent precipitation.
19. The method as claimed in claim 1 to 14 wherein the inoculum is optionally not mixed but added together to the sterilized nutritive matrix.
20. The method as claimed in claim 1 to 14 wherein the enzyme preparation is used for baking for improved freshkeeping, whitening, softening, crumb texture or volume increase.
21. An enzyme preparation for improving baked products obtained from Rhizopus sp. and Aspergillus sp.
22. An enzyme preparation as claimed in claim 21 wherein Rhizopus sp. is Rhizopus oryzae.
23. An enzyme preparation as claimed in claim 22 wherein the Aspergillus sp. is Aspergillus niger.
24. An enzyme preparation as claimed in claim 21 wherein the enzyme preparation is granulated.
25. An enzyme preparation as claimed in any one of claims 21- 24 further comprising a preservative.
26. An enzyme preparation as claimed in claim 25 wherein the preservative is selected from benzoates or sorbates.
27. An enzyme preparation as claimed in any one of the preceding claims further comprising a stabilizer.
28. An enzyme preparation as claimed in claim 27 wherein the stabilizer is selected from inorganic salts, polyols, sugars or their combinations .
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| PCT/IN2001/000094 WO2002088342A1 (en) | 2001-04-30 | 2001-04-30 | An enzyme preparation for improved baking quality and a process for preparing the same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| PCT/IN2001/000094 WO2002088342A1 (en) | 2001-04-30 | 2001-04-30 | An enzyme preparation for improved baking quality and a process for preparing the same |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO2002088342A1 true WO2002088342A1 (en) | 2002-11-07 |
Family
ID=11076345
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/IN2001/000094 WO2002088342A1 (en) | 2001-04-30 | 2001-04-30 | An enzyme preparation for improved baking quality and a process for preparing the same |
Country Status (1)
| Country | Link |
|---|---|
| WO (1) | WO2002088342A1 (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004113490A2 (en) * | 2003-06-24 | 2004-12-29 | HöFer Bioreact GmbH | Directed selective solid-phase culturing of stable microbial mixed populations for the continuous preparation of defined enzyme and metabolite mixture and bioreactor |
| JP2012139150A (en) * | 2010-12-28 | 2012-07-26 | Kitakyushu Foundation For The Advancement Of Industry Science & Technology | Production method of amylase by using mixed bacterial culture of aspergillus bacterium and rhizopus bacterium |
| US8663720B2 (en) * | 2004-03-12 | 2014-03-04 | Nippon Supplement, Inc. | Process for preparing fermented tea and composition |
Citations (6)
| 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 (en) * | 1984-06-30 | 1986-04-11 | Niigataken | Production of 'koji' variant |
| EP0585988A1 (en) * | 1992-07-27 | 1994-03-09 | Gist-Brocades N.V. | Enzyme product and method for improving bread quality |
| EP0913092A2 (en) * | 1997-10-31 | 1999-05-06 | Amano Pharmaceutical Co., Ltd. | Dough composition and preparation thereof |
| WO1999028366A1 (en) * | 1997-12-04 | 1999-06-10 | Board Of Regents Of University Of Nebraska | Degradable polyesters |
| EP1022329A1 (en) * | 1999-01-25 | 2000-07-26 | GIE Agro Industrie | Multi-enzyme product comprising glucoamylolytic, proteolytic and xylanolytic activities, and process to produce it by solid state fermentation of wheat bran by Aspergillus niger |
-
2001
- 2001-04-30 WO PCT/IN2001/000094 patent/WO2002088342A1/en active Application Filing
Patent Citations (6)
| 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 (en) * | 1984-06-30 | 1986-04-11 | Niigataken | Production of 'koji' variant |
| EP0585988A1 (en) * | 1992-07-27 | 1994-03-09 | Gist-Brocades N.V. | Enzyme product and method for improving bread quality |
| EP0913092A2 (en) * | 1997-10-31 | 1999-05-06 | Amano Pharmaceutical Co., Ltd. | Dough composition and preparation thereof |
| WO1999028366A1 (en) * | 1997-12-04 | 1999-06-10 | Board Of Regents Of University Of Nebraska | Degradable polyesters |
| EP1022329A1 (en) * | 1999-01-25 | 2000-07-26 | GIE Agro Industrie | Multi-enzyme product comprising glucoamylolytic, proteolytic and xylanolytic activities, and process to produce it by solid state fermentation of wheat bran by Aspergillus niger |
Non-Patent Citations (9)
| 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)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004113490A2 (en) * | 2003-06-24 | 2004-12-29 | HöFer Bioreact GmbH | Directed selective solid-phase culturing of stable microbial mixed populations for the continuous preparation of defined enzyme and metabolite mixture and bioreactor |
| WO2004113490A3 (en) * | 2003-06-24 | 2005-07-28 | Hoefer Bioreact Gmbh | Directed selective solid-phase culturing of stable microbial mixed populations for the continuous preparation of defined enzyme and metabolite mixture and bioreactor |
| 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 (en) * | 2010-12-28 | 2012-07-26 | Kitakyushu Foundation For The Advancement Of Industry Science & Technology | Production method of amylase by using mixed bacterial culture of aspergillus bacterium and rhizopus bacterium |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP4966346B2 (en) | Method for producing filamentous fungus culture | |
| US3880742A (en) | {62 -1,4,/{62 1,3 Glucanase | |
| JPH06339375A (en) | Glucoamylase enzyme demarcation | |
| CN1323167A (en) | Enzyme-modified cereal suspensions | |
| JPS6251944A (en) | Yeast expanded bread product and its production | |
| CN113388535B (en) | Staple food leavening agent and preparation method and application thereof | |
| John | Amylases-bioprocess and potential applications: a review | |
| JP2021112190A (en) | Method for concentrating protein in grain powder | |
| KR101394009B1 (en) | Process for production of liquid koji | |
| CN101831416B (en) | Pullulanase and production method thereof | |
| US20200305444A1 (en) | Method for preparing food products comprising rye | |
| JPH10120704A (en) | Composition of water-soluble polysaccharides derived from barley malt, its production and use thereof | |
| WO2002088342A1 (en) | An enzyme preparation for improved baking quality and a process for preparing the same | |
| WO2013170440A1 (en) | Bacterial culture media and methods for their preparation and use | |
| US5035903A (en) | High fiber bakery products | |
| Arifin et al. | Profile of microorganisms and amylose content of white corn flours of two local varieties as a affected by fermentation process | |
| US5187081A (en) | Process for preparing protease from endothia parasitica using glucanases to reduce viscosity | |
| Dhingra et al. | Enzymes in Food Industry and Their Regulatory Oversight | |
| WO2020184596A1 (en) | Amylase composition | |
| WO2020184098A1 (en) | Amylase composition | |
| US6379721B1 (en) | Process for preparation of α-amylase from Tinospora cordifolia Miers useful for starch saccharification | |
| RU2294368C1 (en) | Food supplement for hydrolysis of plant raw materials | |
| Kwatia et al. | Hydrolytic activity of amylase produced in solid-state fermentation by a local isolate of Aspergillus niger | |
| KR101733547B1 (en) | - MBF378 Novel Aspergillus oryzae MBF378 with high activity of -amylase and glucoamylase | |
| EP0528612A2 (en) | Amylase capable of digesting raw starch |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| AK | Designated states |
Kind code of ref document: A1 Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT TZ UA UG US UZ VN YU ZA ZW |
|
| AL | Designated countries for regional patents |
Kind code of ref document: A1 Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE TR BF BJ CF CG CI CM GA GN GW ML MR NE SN TD TG |
|
| DFPE | Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101) | ||
| 121 | Ep: the epo has been informed by wipo that ep was designated in this application | ||
| REG | Reference to national code |
Ref country code: DE Ref legal event code: 8642 |
|
| 122 | Ep: pct application non-entry in european phase | ||
| NENP | Non-entry into the national phase |
Ref country code: JP |