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WO1997018295A1 - Rendement accru des levures par utilisation de solutes/osmoprotecteurs compatibles - Google Patents

Rendement accru des levures par utilisation de solutes/osmoprotecteurs compatibles Download PDF

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Publication number
WO1997018295A1
WO1997018295A1 PCT/AU1996/000719 AU9600719W WO9718295A1 WO 1997018295 A1 WO1997018295 A1 WO 1997018295A1 AU 9600719 W AU9600719 W AU 9600719W WO 9718295 A1 WO9718295 A1 WO 9718295A1
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WO
WIPO (PCT)
Prior art keywords
yeast
glycerol
concentration
dough
sugar
Prior art date
Application number
PCT/AU1996/000719
Other languages
English (en)
Inventor
Paul Victor Attfield
David Keith Myers
Stephen Pehm
Vijendran Mario Joseph
Original Assignee
Burns Philp Technology Pty. 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 Burns Philp Technology Pty. Limited filed Critical Burns Philp Technology Pty. Limited
Priority to AU74868/96A priority Critical patent/AU7486896A/en
Publication of WO1997018295A1 publication Critical patent/WO1997018295A1/fr

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Classifications

    • 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
    • C12N1/16—Yeasts; Culture media therefor
    • A—HUMAN NECESSITIES
    • A21—BAKING; EDIBLE DOUGHS
    • A21D—TREATMENT OF FLOUR OR DOUGH FOR BAKING, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS
    • A21D6/00—Other treatment of flour or dough before baking, e.g. cooling, irradiating or heating
    • A21D6/001—Cooling
    • 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
    • A21D8/047—Methods for preparing dough; Treating dough prior to baking treating dough with microorganisms or enzymes with yeasts

Definitions

  • yeasts Many industrial applications of yeast involve exposure to high osmotic pressures, exerted by various ionic and non-ionic chemicals or solutes (eg. salts and sugars).
  • baker's yeast may be exposed to high osmotic pressures in production (eg., during fermentation and drying or freezing) and application (eg., plain and sugar doughs, frozen doughs).
  • industrial yeasts in general may be subjected to high osmotic pressures in fermentations involving, for example, production of alcohol (potable and non-potable), soy sauce, miso etc.
  • High external osmotic pressures result in removal of water from cells and increased concentration of internal solutes.
  • glycerol as compatible solutes in osmoregulation of yeasts and other organisms in hyperosmotic environments (for example, see 3- 8).
  • truly salt- and osmo-tolerant yeast eg., Debaryomyces hansenii, Zygosaccharomyces rouxii, Torulaspora delbruckii
  • Saccharomyces cerevisiae baker's yeast which are not considered truly osmotolerant strains, show increased glycerol production but also leak much of this from the cell (5-8).
  • the present inventors have found that the performance of yeast for high osmotic processes can be improved by the addition of compatible solutes such as glycerol to yeast biomass. As will be understood by those experienced in this area there are a number of factors in assessing the performance of yeast. These include, for example, fermentative activity, leavening activity, time taken to proof doughs and retention of activity during storage. Improvement in any one these criteria would be viewed as an increase in performance.
  • the present invention consists in a method of increasing the performance of yeast the method comprising exposing the yeast to at least one compatible solute/osmoprotectant for a period of time sufficient to result in an intracellular concentration of the at least one compatible solute/osmoprotectant of at least 70 ⁇ Moles/gram dry veast.
  • the yeast is exposed to the at least one compatible solute/osmoprotectant for a period of time sufficient to result in an intracellular concentration of the at least one compatible solute/osmoprotectant of at least lOO ⁇ Moles/gram dry yeast.
  • the compatible solutes/osmoprotectants is preferably selected from the group consisting of glycerol, trehalose, sucrose, maltose, glucose, fructose, mannose, ammonium salts, amino compounds, and combinations thereof.
  • the most preferred compatible solute/osmoprotectant is glycerol. Where the solute is glycerol it is preferred that the yeast is exposed to the glycerol for a sufficient period of time to result in an intracellular concentration of glycerol of at least lOO ⁇ Moles/gram dry yeast equivalents.
  • the yeast is exposed to glycerol at a concentration of at least 0.1M and preferably at a concentration of at least 0.2M.
  • concentration of glycerol is at least 0.4M.
  • the yeast is exposed to the compatible solute for at least 24 hours.
  • the yeast is in the form of yeast having greater than 18% dry yeast material.
  • the present invention consists in a yeast product produced by the method of the present invention, preferably a cream yeast or compressed yeast product.
  • the present invention consists in a frozen dough product including a yeast prepared by the method of the present invention.
  • the present invention consists in a cream yeast product characterised in that the yeast has an internal glycerol concentration of at least lOO ⁇ Moles/gram dry yeast equivalents.
  • the present invention consists in compressed yeast product characterised in that the yeast has an internal glycerol concentration of at least lOO ⁇ Moles/gram dry yeast equivalents.
  • Figures 1 and 2 show uptake of glycerol by yeast stored at 4°C (•, yeast with no added glycerol: ⁇ . ⁇ , ⁇ yeast plus 0.2M glycerol added). Significant benefit from glycerol addition is not observed until at least 24 hr contact time, therefore optimal application would be by stirring into cream at chilled temperatures and mixing gently for preferably >24hr.
  • Figure 3 shows CO z gas -producing activity and stability (retention of activity with storage time) of yeast cream in 16% sugar doughs (O, yeast plus 0.2M glycerol; •, yeast with no added glycerol). Benefit is observable in sugar dough application of cream yeast where the level of glycerol proposed gives marked improvement in stability of product.
  • Figure 4 gives the activity and stability of compressed yeast made from glycerol-treated or non-glycerol-treated cream (O, yeast plus 0.2M glycerol; •, yeast with no added glycerol). Benefit is observable in sugar dough application of compressed yeast made from glycerol-treated cream where the level of glycerol proposed gives marked improvement in stability of product.
  • Figure 5 shows plain dough activity and stability of yeast cream in plain doughs (O, yeast plus 0.2M glycerol; •, yeast with no added glycerol). Benefit is observable in plain dough application of cream yeast where the level of glycerol proposed gives improvement in activity and some improvement in stability of product.
  • Figure 6 shows plain dough activity and stability of compressed yeast made from glycerol -treated or non-glycerol-treated cream yeast
  • FIG. 7 shows proof times for 10% sugar frozen doughs that have been stored at -21°C for up to 8 weeks.
  • A Yeast at 4°C for one day
  • B Yeast at 4°C for three days
  • C Yeast at 4°C for seven days
  • D Yeast at 4°C for twenty one days.
  • ⁇ doughs made with yeast containing no added glycerol
  • ⁇ doughs made with yeast to which glycerol was added at 0.2M
  • A doughs made with yeast to which glycerol was added at 0.3M).
  • Figure 8 gives proof times for 10% sugar frozen doughs that have been stored at -21°C for up to 8 weeks.
  • A Yeast at 4°C for one day
  • B Yeast at 4°C for three days
  • C Yeast at 4°C for seven days
  • D Yeast at 4°C for twenty one days.
  • ⁇ doughs made with yeast containing no added glycerol
  • ⁇ doughs made with yeast to which glycerol was added at 0.4M.
  • Glycerol (food grade) was obtained from Henkel Australia. Bakers' flour was purchased from Defiance mills (Sydney, Australia); sugar from CSR (Sydney, Australia); Shortening was DPS2; Bread improver was Bakerine Special for activity testing and Bakedoh for Bake testing - both obtained from Mauri Integrated Ingredients (Sydney, Australia). Salt was Analar. Industrially produced yeast was obtained from Burns Philp & Co. Ltd. factories.
  • Such yeast preparations can be taken and glycerol mixed in, preferably whilst maintaining a cool temperature of the yeast ( ⁇ 10°C, and preferably ⁇ 4°C). Mixing time is as rapid as possible whilst achieving as homogenous mixing as possible.
  • Yeast can then be stored ( ⁇ 10°C, and preferably ⁇ 4°C) as crumble to allow glycerol to equilibrate across membranes prior to making into blocks, or use as crumble in its own right.
  • Compressed yeast was suspended in water then added to the dry ingredients, together with the salt solution. The complete dough was mixed for three minutes, reaching a final dough temperature of 30°C. The finished dough was transferred to a standard Fermentograph tin then put into the SJA apparatus. Tests were carried out for two hours knocking the dough down after the first hour.
  • the finished dough was transferred to a standard Fermentograph tin then put into the SJA apparatus. Tests were carried out for two hours knocking the dough down after the first hour.
  • Glycerol was added to cream yeast as described previously and compressed yeast samples prepared. These were stored refrigerated for up to 21 days and at intervals yeast was used for preparing frozen doughs.
  • the dough composition used is set out in Table 5.
  • Mixing time was determined by Farinograph and water absorption by Extensograph. Ingredients were weighed into a bucket, other components (except water) added so as to not come into contact with each other, mixed for 1 minute and then water added. The dough was mixed until fully developed. Final dough temperatures were 20 +/-2°C. Seven 520g dough pieces were produced and moulded (six inch Mono Moulder), one being tested immediately for proof time (time to rise to 120 mm) and the other six were blast frozen at -40°C until the core temperature was — -5°C. Frozen doughs were then stored at -21°C for 1, 4 and 8 weeks: two doughs were defrosted (at 4°C for 16 hours prior to proof testing) and tested at each time.
  • Figures 7 and 8 show that addition of glycerol to yeast cream followed by a sufficient period for glycerol to enter yeast cells and subsequent preparation of compressed yeast significantly improved stability of 10% sugar dough over eight weeks storage at -21°C.
  • the proof times of the resulting frozen 10% sugar doughs increased only slightly from 1 week to 8 weeks storage at -21°C.
  • Untreated yeast suffered an increase in proof time due to initial freezing and especially 0.4 M glycerol additions significantly reduced the loss on freezing. Benefits were related to the amount of glycerol added in the range 0.2 to 0.4 M (total water basis).
  • the 0.2 M and 0.3 M treatments gave similar results for doughs made after 1, 7 and 14 days storage of the yeast at 4°C, but after 21 days storage the 0.3 M addition gave significantly better results than the 0.2 M addition.
  • 0.4 M glycerol addition resulted in significantly better frozen 10% sugar doughs than the 0.2 M or 0.3 M treatments.
  • the performance of the treated material relative to the control became greater as storage time of the yeast (as either cream or compressed at 4°C) prior to dough production progressed: the longer untreated yeast was kept at 4°C prior to frozen 10% sugar dough production, the worse the keeping of the activity (at -21°C) became - this occurred to a lesser extent for the 0.2 and 0.3 M treated yeast and did not occur significantly for the 0.4 M glycerol treated yeast.
  • yeast biomass is given a "head start' in subsequent osmotically stressing applications.
  • an osmoprotectant/ compatible solute such as glycerol
  • glycerol-treated yeast achieve a crucial level of osmotic equilibration earlier than non-glycerol-treated yeast cells. Accordingly, in the case of glycerol, a natural osmoprotectant made by many yeast strains, it is important to allow added glycerol to soak into (or be taken up by) yeast cells so that they are pre-loaded with a significant concentration of protectant prior to industrial application.
  • yeast Because the basal content of osmoprotectant in yeast product is not normally high enough to cope with immediate osmostress when mixed into a sugar dough, yeast has to make glycerol and so sits in a lag phase, during which little gas production occurs, until it achieves a critical concentration that provides osmotic stability. Only then does gas production become significant. Hence addition of glycerol with enough time to allow this to equilibrate across the yeast membrane, should give the yeast a "head start" by reducing the amount of intrinsic glycerol manufacture to reach the critical concentration.
  • strains of yeast may be engineered or selected (by recombinant technology or breeding/ fusion) for improved glycerol production and retention at, or in excess of, say, >0.4 mMol glycerol/ gram dry yeast within 30 min in a high sugar dough system, preferably with as made invertase levels being below 10 units of activity.
  • One such approach might be to boost the level of glycerol-3-phosphate dehydrogenase in yeasts as this is induced by salt and sugar stress and forms part of the high osmotic glycerol response (10,11).
  • Another approach is to boost the levels of substrates intracellularly available for glycerol synthetic enzymes.
  • Yet another way to increase glycerol production in yeast is to induce this process by treatment with salts or other osmolytes, which is a well known phenomenon and is already used in the industry (12,13).
  • any manipulation would also benefit from having cells modified to retain glycerol in non-osmotic stress conditions, e.g. manipulation of glycerol channel/porter proteins.
  • GPDl which encodes glycerol-3-phosphate dehydrogenase, is essential for growth under osmotic stress in Saccharomyces cerevisiae, and its expression is regulated by the high-osmolarity glycerol response pathway. Molecular and Cellular Biology, 14, 4135-4144.

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Abstract

La présente invention concerne un procédé d'accroissement du rendement des levures consistant à exposer la levure à au moins un soluté ou osmoprotecteur pendant une durée suffisante à l'obtention d'une concentration intracellulaire d'au moins un soluté ou osmoprotecteur dans la levure sèche d'au moins 70 νmoles/g. On préfère une durée d'exposition suffisante pour l'obtention d'une concentration intracellulaire de ce soluté ou de cet osmoprotecteur dans la levure sèche d'au moins 100 νmoles/g. Les solutés ou osmoprotecteurs compatibles sont de préférence le glycérol.
PCT/AU1996/000719 1995-11-14 1996-11-13 Rendement accru des levures par utilisation de solutes/osmoprotecteurs compatibles WO1997018295A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU74868/96A AU7486896A (en) 1995-11-14 1996-11-13 Improved yeast performance using compatible solutes/osmoprotectants

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
AUPN6548A AUPN654895A0 (en) 1995-11-14 1995-11-14 Improved yeast performance
AUPN6548 1995-11-14

Publications (1)

Publication Number Publication Date
WO1997018295A1 true WO1997018295A1 (fr) 1997-05-22

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6770360B2 (en) 1998-06-12 2004-08-03 Avery Dennison Corporation Multilayered thermoplastic film and sign cutting method using the same

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0435606A2 (fr) * 1989-12-26 1991-07-03 Ichinobe Baking Co. Ltd Pâte à pain

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0435606A2 (fr) * 1989-12-26 1991-07-03 Ichinobe Baking Co. Ltd Pâte à pain

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
DERWENT ABSTRACT, Accession No. 89-316351, Class D16; & CS,A,87 08359, (BREIEROVA E.), 12 September 1989. *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6770360B2 (en) 1998-06-12 2004-08-03 Avery Dennison Corporation Multilayered thermoplastic film and sign cutting method using the same

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Publication number Publication date
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