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WO1991018977A1 - Procede de preparation d'une composition de pentosanase active dans les pates de boulangerie - Google Patents

Procede de preparation d'une composition de pentosanase active dans les pates de boulangerie Download PDF

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Publication number
WO1991018977A1
WO1991018977A1 PCT/EP1991/000938 EP9100938W WO9118977A1 WO 1991018977 A1 WO1991018977 A1 WO 1991018977A1 EP 9100938 W EP9100938 W EP 9100938W WO 9118977 A1 WO9118977 A1 WO 9118977A1
Authority
WO
WIPO (PCT)
Prior art keywords
fermentation medium
net
preparation
baking
xylan
Prior art date
Application number
PCT/EP1991/000938
Other languages
German (de)
English (en)
Inventor
Joachim Schindler
Hans Friedrich Pfeiffer
Horst-Werner Wollenweber
Heinrich Waldhoff
Wolfgang Adams
Original Assignee
Henkel Kommanditgesellschaft Auf Aktien
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 Henkel Kommanditgesellschaft Auf Aktien filed Critical Henkel Kommanditgesellschaft Auf Aktien
Publication of WO1991018977A1 publication Critical patent/WO1991018977A1/fr

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
    • C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
    • C12Y302/01032—Xylan endo-1,3-beta-xylosidase (3.2.1.32), i.e. endo-1-3-beta-xylanase
    • A—HUMAN NECESSITIES
    • A21—BAKING; EDIBLE DOUGHS
    • A21D—TREATMENT OF FLOUR OR DOUGH FOR BAKING, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS
    • 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
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
    • C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
    • C12N9/2477—Hemicellulases not provided in a preceding group
    • C12N9/248—Xylanases
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
    • C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
    • C12Y302/01008—Endo-1,4-beta-xylanase (3.2.1.8)

Definitions

  • the invention relates to a fermentation process for producing a pentosanase preparation with increased baking activity.
  • Pentosanases are enzymes that are able to break down pentosans.
  • the pentosans belong to the henricelluloses.
  • the most important component is arabinoxylosan, which chemically consists of a chain-shaped xylan (polymer of D-xylose) with arabinose in the side chains.
  • Such poiysaccharides can also be found as fiber in cereal flours.
  • US Pat. No. 3,512,992 therefore proposes to use pentosanases and, in particular, xylanases in the bakery when preparing dough. It is known that the dough viscosity can be reduced by using pentosanase. Furthermore, white bread becomes particularly soft and old baking can be delayed. In the case of rye pastries, the elasticity of the crumbs can be improved (C. Garns "The use of microbial enzymes in the bakery in cereals, flour and bread", 30th year, volume 5, May 1976, page 113 ff.). An overview-like description of xylanases and their properties can be found in J. Woodward in "Topics in Enzymology and Fermentation, Biotechnology, Vol. 8 (1984), page 9 ff.
  • inducers such as ⁇ -methyl-xyloside
  • xylan-degrading microorganisms form more than just one type of xylanase, which differ, inter alia, in that they are back-active or back-inactive. It has been shown in food technology practice that the use of pentosanases or Volume enlargement of a biscuit to be achieved correlated directly with a measurable xylanase activity of the baking active xylanase. There is therefore a need for a process which specifically allows enzyme preparations with high baking activity to be produced.
  • the invention therefore relates to a process for producing a baking-active pentosanase preparation, in which microorganisms which are capable of forming pentosanases are grown in a fermentation medium, the cell mass is separated off at the end of the culture period and the enzyme preparation is removed the fermenter broth wins, characterized in that, in addition to or instead of xylan and / or xylan-containing mixtures, the substance ⁇ -methylxyloside is added in quantities of 0.01 to 5% by weight, based on the fermentation medium, to induce baking activity in the fermentation medium.
  • the process according to the invention is suitable for the production of baking-active pentosanase preparations from a large number of microorganism strains of the most diverse genera, for example Aspergillus Rhizopus, Trichodema Bacillus and the like.
  • strains of the genus Aspergillus are preferred, especially Aspergillus niger and Aspergillus awamori and among these Aspergillus awamori DSM 5937.
  • the effect on which the invention is based is that the substrate ⁇ -methylxyloside, as substrate analogs, stimulates the microorganisms in question for the synthesis of pentosanases (xylanases) - in particular also for the formation of the back-active enzyme.
  • the essence of the invention is therefore to add ⁇ -methylxyloside to the fermentation medium in the preparation of the enzyme preparations. Suitable addition amounts are between 0.01 and 5% by weight, in particular between 0.01 and 1% by weight.
  • ⁇ -methylxyloside can be used together with xylan or instead of xylan. It is preferred to use the substance instead of xylan. It is also possible to use ⁇ -methylxyloside together with a component containing xylane, for example together with rye bran, or preferably to add it instead of such a component.
  • Ss-Methylxyloside not only achieves higher xylanase yields, but also significantly reduces the fermentation time until maximum xylanase activity is reached.
  • the fermentation media to be used according to the invention have a composition which is customary in the production of enzymes by microorganisms of the genus Aspergillus, in particular Aspergillus niger or Aspergillus awamori.
  • the fermentation media contain a C source, an N source and trace elements.
  • Products containing polysaccharide are used as the C source, for example corn starch or degraded corn starch or another starch source.
  • Soy flour, but also corn steep liquor can be used as the N source.
  • ammonium salts such as ammonium nitrate
  • the substance used as the C source is present in the fermentation medium in an amount of 0.2 to 5% by weight. The same applies to protein-like substances.
  • the Feraenta tion medium nor the usual trace elements, such as salts of potassium, sodium, magnesium, manganese, iron and the like.
  • the amount of trace elements is less than 1% by weight and is about 0.01 to 0.2% by weight per type of cation, based on the particular cation.
  • the trace elements are preferably present as soluble salts, for example as phosphates, nitrates or sulfates.
  • the combinations are chosen so that precipitation is avoided.
  • the total pH of the fermentation medium is slightly acidic to neutral and can take on values between pH 4.5 and pH 7.5, in particular around pH 6.0.
  • the fermentative production of the pentosanases can be carried out in customary fermentation plants, working under aerobic conditions at room temperature or slightly above.
  • the culture lasts from 24 hours to a few days, and the xylanase formation can be monitored analytically.
  • the fungal mycelium is first separated from the fermenter broth by filtration.
  • the liquid phase obtained in this way can be processed further in various processes. It is thus possible to precipitate the enzyme from this liquid phase, for example with ammonium sulfate, and to further purify or process the precipitated enzyme directly.
  • the pentosanase concentrates produced according to the invention show high pentosanase (xylanase) activity with simultaneously improved baking activity.
  • the xylanase activity can be determined using the dinitrosalicylic acid method as follows:
  • the enzyme-containing solution is incubated in acetate-buffered solution in the presence of approx. 1% xylan at 40 ° C. for 15 minutes. Then an alkaline dinitrosalicylic acid DNA solution (7g DNA, 12g NaOH, 200g potassium sodium tartrate, 5.5g phenol, 5g Na2S2 ⁇ s per 1 l H2O) is added and the color development at 540 nm compared to a xylose standard is measured. The enzyme unit 1 U is present if in 1 min. fragments are released from the xylan under the reaction conditions, the reduction equivalents of which correspond to 1 ⁇ mol of xylose.
  • the increase in volume of rolls is measured from a defined amount of a standard dough. To do this, measure the length, width and height of 30 such buns, add the dimensions in cm and subtract from the number obtained the dimensions of likewise 30 buns which were produced from the same amount of the same dough without the enzyme addition. Examples
  • Vaccine culture To produce vaccine cultures, well-transported agar cultures of strain 5937 (XY1-A007) were washed off with 0.1 PO / j buffer (pH 6.5), the spore suspension was filtered over sterile glass wool, with 10% glycerol as a protective reagent added and stored in portions at -25 ° C.
  • the fungal mycelium was separated by filtration and the cell-free culture filtrate was purified by dialysis and concentrated by ultrafiltration. A freeze-dried powder was obtained from the concentrate, which was mixed with soy flour to a product with 15000 U / g.
  • a dough was produced from 1000 g of wheat flour, type 550, 20 g of salt, 60 g of yeast, 580 g of water and 30 g of baking agent with or without the addition of enzyme. A total of 5 min. kneaded. After a rest period of 5 min. exactly 1500 g are weighed out, this section is kneaded manually and in the following bale oven for a further 10 min. rested.
  • the bale is divided into 50 g dough pieces and knitted round.
  • the round dough pieces are after 11/2 min. Rest time screwed into a »long-knitting machine.
  • Example 1 was repeated, but xylan or rye flour bran was used instead of ⁇ -methylxylitol. The results are shown in the table.

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  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Food Science & Technology (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Bakery Products And Manufacturing Methods Therefor (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

Selon un procédé de préparation d'une composition de pentosanase, on cultive un microorganisme capable de former des pentosanases dans un milieu de fermentation. Afin d'augmenter l'activité dans les pâtes des enzymes ainsi obtenus, on ajoute au milieu de fermentation, en plus ou à la place du xylane et/ou des mélanges contenant du xylane, des quantités comprises entre 0,01 et 5 % en poids, par rapport au milieu de fermentation, de la substance beta-méthylxyloside, afin d'induire l'activité dans les pâtes de boulangerie.
PCT/EP1991/000938 1990-05-28 1991-05-21 Procede de preparation d'une composition de pentosanase active dans les pates de boulangerie WO1991018977A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE19904017150 DE4017150A1 (de) 1990-05-28 1990-05-28 Verfahren zur herstellung eines backaktiven pentosanase-praeparates
DEP4017150.7 1990-05-28

Publications (1)

Publication Number Publication Date
WO1991018977A1 true WO1991018977A1 (fr) 1991-12-12

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Country Status (3)

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DE (1) DE4017150A1 (fr)
IE (1) IE911797A1 (fr)
WO (1) WO1991018977A1 (fr)

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002003805A1 (fr) 2000-07-06 2002-01-17 Novozymes A/S Procede de preparation d'une pate ou d'un produit de cuisson a partir d'une pate avec apport d'enzymes lipolytiques
WO2004099400A2 (fr) 2003-05-09 2004-11-18 Novozymes A/S Enzymes lipolytiques variants
EP2428572A2 (fr) 2007-03-09 2012-03-14 Danisco US, Inc., Genencor Division Variants de l'alpha-amylase d'une espèce de Bacillus alcaliphile, compositions comprenant des variants de l'alpha-amylase, et procédés d'utilisation
US8318157B2 (en) 2007-03-14 2012-11-27 Danisco Us Inc. Trichoderma reesei α-amylase enhances saccharification of corn starch
US9040279B2 (en) 2008-06-06 2015-05-26 Danisco Us Inc. Saccharification enzyme composition and method of saccharification thereof
US9040278B2 (en) 2008-06-06 2015-05-26 Danisco Us Inc. Production of glucose from starch using alpha-amylases from Bacillus subtilis
EP3133154A2 (fr) 2009-05-19 2017-02-22 DuPont Nutrition Biosciences ApS Polypeptides d'amylase

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE69228378T2 (de) * 1991-04-02 1999-08-26 Novo Nordisk A/S Xylanase, entsprechende rekombinante dna-sequenz, xylanase enthaltendes agens, und seine verwendung
CN102978258A (zh) 2008-01-02 2013-03-20 丹尼斯科美国公司 应用嗜糖假单胞菌g4-淀粉酶和其变体获得乙醇的无葡糖淀粉酶的方法
CN102016044B (zh) 2008-04-30 2014-11-26 丹尼斯科美国公司 新型嵌合α-淀粉酶变体
MX2010013114A (es) 2008-06-06 2011-04-05 Danisco Inc Alfa-amilasas variantes de bacillus subtilis y metodos de uso de las mismas.

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3512992A (en) * 1968-04-02 1970-05-19 Delmar Chem Baking additive and method for producing baked goods
JPS5257393A (en) * 1975-11-01 1977-05-11 Mitsui Seitou Kk Production of actinomycetes xylanase by betaa heteroxyloside

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3512992A (en) * 1968-04-02 1970-05-19 Delmar Chem Baking additive and method for producing baked goods
JPS5257393A (en) * 1975-11-01 1977-05-11 Mitsui Seitou Kk Production of actinomycetes xylanase by betaa heteroxyloside

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
Chemical Abstracts, Band 103, 28. Oktober 1985 (Columbus, Ohio, US) J. Defaye et al.:"Induction of D-xylan-degrading enzymes in Trichoderma lignorum by nonmetabolizable inducers. A synthesis of 4-thioxylobiose", siehe Seite 756 *
Chemical Abstracts, Band 104, 23. Juni 1986 (Columbus, Ohio, US) A.M. Deschamps et al.: "Comparitive studies on factors regulating xylanase synthesis in Bacillus and Corynebacterium spp.", siehe Seite 330 *
Chemical Abstracts, Band 110, 5. Juni 1989 (Columbus, Ohio, US) S.R. Biswas et al.:"Induction of xylanase in Aspergillusochraceus", siehe Seite 290 *
Chemical Abstractsa, Band 110, 22. Mai 1989 (Columbus, Ohio, US) M.J. Bailey et al.: "Production of xylanolytic enzymes by stains of Aspergillus", siehe Seite 605 *
Getreide Mehl und Brot, Band 30, Nr. 5, Mai 1976 T.C. Gams: "Der Einsatz von mikrobiellen Enzymen in der B{ckerei", Seiten 113-116 *
Japanese Patents Gazette, Band 77, Nr. 25, Sektion CH: Chemical, 1. August 1977 (Londen, GB) siehe B9, Zusammenfassung Nr. 44421y/25 & JP-A-52057393 (MITSUI SUGAR K.K.) 11. Mai 1977 *

Cited By (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002003805A1 (fr) 2000-07-06 2002-01-17 Novozymes A/S Procede de preparation d'une pate ou d'un produit de cuisson a partir d'une pate avec apport d'enzymes lipolytiques
WO2004099400A2 (fr) 2003-05-09 2004-11-18 Novozymes A/S Enzymes lipolytiques variants
EP2270140A2 (fr) 2003-05-09 2011-01-05 Novozymes A/S Enzymes lipolytiques variants
EP2270139A2 (fr) 2003-05-09 2011-01-05 Novozymes A/S Enzymes lipolytiques variants
EP2290057A2 (fr) 2003-05-09 2011-03-02 Novozymes A/S Enzymes lipolytiques variants
EP2428572A2 (fr) 2007-03-09 2012-03-14 Danisco US, Inc., Genencor Division Variants de l'alpha-amylase d'une espèce de Bacillus alcaliphile, compositions comprenant des variants de l'alpha-amylase, et procédés d'utilisation
US8318157B2 (en) 2007-03-14 2012-11-27 Danisco Us Inc. Trichoderma reesei α-amylase enhances saccharification of corn starch
US8916369B2 (en) 2007-03-14 2014-12-23 Danisco Us Inc. Trichoderma reesei α-amylase is a maltogenic enzyme
US9040279B2 (en) 2008-06-06 2015-05-26 Danisco Us Inc. Saccharification enzyme composition and method of saccharification thereof
US9040278B2 (en) 2008-06-06 2015-05-26 Danisco Us Inc. Production of glucose from starch using alpha-amylases from Bacillus subtilis
EP3133154A2 (fr) 2009-05-19 2017-02-22 DuPont Nutrition Biosciences ApS Polypeptides d'amylase
EP3346004A1 (fr) 2009-05-19 2018-07-11 DuPont Nutrition Biosciences ApS Polypeptides d'amylase
EP3412771A1 (fr) 2009-05-19 2018-12-12 DuPont Nutrition Biosciences ApS Polypeptides d'amylase
EP3473711A1 (fr) 2009-05-19 2019-04-24 DuPont Nutrition Biosciences ApS Polypeptides d'amylase
EP3591046A1 (fr) 2009-05-19 2020-01-08 DuPont Nutrition Biosciences ApS Polypeptide amylase
EP3620518A1 (fr) 2009-05-19 2020-03-11 DuPont Nutrition Biosciences ApS Polypeptide d'amylase
EP3783103A1 (fr) 2009-05-19 2021-02-24 DuPont Nutrition Biosciences ApS Polypeptides d'amylase

Also Published As

Publication number Publication date
DE4017150A1 (de) 1991-12-05
IE911797A1 (en) 1991-12-04

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