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    BMM-S12-10 RD of Yeast

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    Riyan
    The document describes a test to determine respiratory deficiency in yeast cells. Yeast cells that lack certain respiratory enzymes will appear as small white colonies when overlaid with triphenyl tetrazolium chloride (TTC) agar, while respiratory sufficient cells will appear pink to red. To perform the test, TTC agar is prepared according to standard methods and poured over yeast colonies on agar plates. The plates are incubated for one hour at 30°C. Respiratory deficient colonies remain white while sufficient colonies turn pink to red. The percentage of respiratory deficiency is calculated by counting colonies.

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    BMM-S12-10 RD of Yeast

    Uploaded by

    Riyan
    18 views2 pages
    The document describes a test to determine respiratory deficiency in yeast cells. Yeast cells that lack certain respiratory enzymes will appear as small white colonies when overlaid with triphenyl tetrazolium chloride (TTC) agar, while respiratory sufficient cells will appear pink to red. To perform the test, TTC agar is prepared according to standard methods and poured over yeast colonies on agar plates. The plates are incubated for one hour at 30°C. Respiratory deficient colonies remain white while sufficient colonies turn pink to red. The percentage of respiratory deficiency is calculated by counting colonies.

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    The document describes a test to determine respiratory deficiency in yeast cells. Yeast cells that lack certain respiratory enzymes will appear as small white colonies when overlaid with triphenyl tetrazolium chloride (TTC) agar, while respiratory sufficient cells will appear pink to red. To perform the test, TTC agar is prepared according to standard methods and poured over yeast colonies on agar plates. The plates are incubated for one hour at 30°C. Respiratory deficient colonies remain white while sufficient colonies turn pink to red. The percentage of respiratory deficiency is calculated by counting colonies.

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    © All Rights Reserved
    Download as DOC, PDF, TXT or read online from Scribd
    Download as doc, pdf, or txt
    18 views2 pages

    BMM-S12-10 RD of Yeast

    Uploaded by

    Riyan
    The document describes a test to determine respiratory deficiency in yeast cells. Yeast cells that lack certain respiratory enzymes will appear as small white colonies when overlaid with triphenyl tetrazolium chloride (TTC) agar, while respiratory sufficient cells will appear pink to red. To perform the test, TTC agar is prepared according to standard methods and poured over yeast colonies on agar plates. The plates are incubated for one hour at 30°C. Respiratory deficient colonies remain white while sufficient colonies turn pink to red. The percentage of respiratory deficiency is calculated by counting colonies.

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    © All Rights Reserved
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    of 2

    The South African Breweries Section : 12.

    10
    Page : 1 of 2

    BREWING MANUAL Revision : Draft Rev


    Date : Aug 2002
    Vol. VIII – Microbiological Methods
    Section 12: Techniques:
    12.10 – Respiratory deficiency of Yeast

    1 INTRODUCTION 5 APPARATUS

    a) Respiratory deficient yeast cells are those  Water-bath.

    cells that lack certain respiratory enzymes.  Hot-plate/ magnetic stirrer.

    b) The often appear as small colonies on a  Weighing balance.


    solid Agar medium, and they are associated  Measuring cylinder.
    with poor growth and unfavourable flavour  Stirrer bar.
    production during fermentation.  Spatula.
    c) They also produce lower yeast viabilities.  Glass beaker.
    d) The test for determining respiratory
    deficient cells relies on the fact that if 6 PROCEDURE
    respiratory enzymes are present in yeast
    cells, then yeast is respiratory sufficient, a) Prepare TTC agar as per MMM
    therefore the colourless tetrazolium salt 9.14.Attemperate to 50°C in the water-bath.
    would be reduced to the insoluble form, b) Attemperate to 50°C in the water-bath.
    triphenylformazan and form a red pigment. c) Pour onto MYGP agar plates such that yeast
    colonies are completely overlaid.
    2 HEALTH d) Incubate in a 30°C incubator for 1 hour.
    AND SAFETY
    Health & Safety: Section 3 7. INTERPRETATION
    3 REFERENC
    ES Normal respiratory sufficient colonies turn
    pink to red.
    4 MEDIA Respirtary deficient colonies remain white.
    AND REAGENTS

    a) Triphenyl tetrazolium chloride agar


    (TTC) plates (MMM 9.14).
    b) MYGP agar plates (MMM 9.7) with yeast
    colonies.
    c) Distilled water.
    The South African Breweries Section : 12.10
    Page : 2 of 2

    BREWING MANUAL Revision : Draft Rev


    Date : Aug 2002
    Vol. VIII – Microbiological Methods
    Section 12: Techniques:
    12.10 – Respiratory deficiency of Yeast

    8. Calculation vii. Was the calculation done correctly?

    Total number of what colonies*100


    Total number of colonies (pink/red and
    white)
    = % respiratory deficiency.

    NOTE: TTC cannot be stored.


    It must be used immediately;
    therefore it is advisable to make it up
    in small quantities, depending on the
    number of plates to be overlaid.

    9 HOUSEKEEPING

    All used glassware must be removed


    immediately from the micro laboratory.

    10 CHECKLIST OF CRITICAL AREAS

    i. Were the
    ingredients weighed correctly?
    ii. Were they soaked for 10 minutes
    before heating?
    iii. Were they held
    at the boil for one minute?
    iv. Was the
    attemperation temperature correct?
    v. Were the overlaid yeast colonies
    completely covered?
    vi. Was the
    incubation temperature and time
    correct?

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