1. YEAST BIOCHEMISTRY BIOLOGY BIOTECHNOLOGY
MBAA-Rocky Mountain District Meeting
YEAST
BIOCHEMISTRY
BIOLOGY
BIOTECHNOLOGY By
Professor, Dr. Jim Mattoon
Director, Center for Biotechnology and Bioinformatics
1090 Garlock Lane, Colorado Springs, Colorado 80918
(719) ;
September 15, 2007

2. Alcoholic fermentation by
YEAST BIOCHEMISTRY
Eduard Buchner 1897
Alcoholic fermentation by
cell-free extracts of yeast.
“zymase”

3. ZYMASE NOT A SINGLE ENZYME A MULTIENZYME SYSTEM
EACH ENZYME CATALYZES A SINGLE CHEMICAL CHANGE IN A CHAIN OF REACTIONS NOW CALLED GLYCOLYSIS

4. GLYCOLYSIS INVOLVES MANY ENYMES
AIDED BY COENZYMES DERIVED FROM VITAMINS
SIMILAR ENZYMES AND COENZYMES FOUND IN MUSCLE EXTRACTS
MUCH OF BIOCHEMISTRY IS UNIVERSAL IN LIVING ORGANISMS.

5. COMPARE YEAST & MUSCLE GLYCOLYSIS
START: SUGAR
END: ETHANOL + CO2
MUSCLE
START: SUGAR
END: LACTIC ACID

6. FATHERS OF MICROBIOLOGY
LOUIS PASTEUR – FRANCE
ALCOHOLIC FERMENTATION CAUSED BY LIVING YEAST CELLS
ROBERT KOCH – GERMANY
MANY DISEASES CAUSED BY LIVING CELLS OF BACTERIA

7. YEAST BIOLOGY SINGLE CELL REPRODUCES BY BUDDING
GROWS WITH OR WITHOUT AIR
HAS A TRUE NUCLEUS WITH MULTIPLE LINEAR CHROMOSOMES
HAS A TOUGH CELL WALL
MUCH LARGER THAN BACTERIA

8. VITAMIN DISCOVERY WITH YEAST
RIBOFLAVIN
FOLIC ACID
BIOTIN
COENZYME FOR RESPIRATION
CURE FOR LARGE CELL ANEMIA
BIOASSAY - ALLOWED PURIFICATION OF FOLIC ACID

9. YEAST BIOLOGY ANATOMY & CELL BIOLOGY REPRODUCTION NUTRITION LIFE CYCLE
GENETICS
TRANSFORMATION

11. FUNCTIONS OF CELL COMPONENTS
NUCLEUS: CHROMOSOMES [Strings of genes that control inheritance-DNA].
VACUOLE: Recycling center
CYTOPLASM:
Enzymes: Catalytic proteins controlling metabolism, including fermentation
Ribosomes: Protein factories

12. YEAST GENETICS I
Founded by Øjvind Winge working at the laboratories of the Carlsberg Brewery in Copenhagen in the late 1930s and early 1940s.
He was interested in genes that controlled fermentation, particularly genes that controlled maltose fermentation.
During mashing of malt, the main sugar produced is maltose.

13. LIFE CYCLE - PLOIDY

15. YEAST GENETICS II
He first had to work out the yeast life-cycle & micro dissection methods.
Sexual reproduction
Double chromosomes [Diploids 2n--->4n]
Pairs separate, then separate again. 4n---> n+n+n+n. 4 haploid spores
Spores germinate & budding follows
Genes segregate in pairs.
For example mating type;2 α : 2 αlpha.
Different forms of same gene also 2:2.

16. YEAST GENETICS III
During the next 50 years over 1000 genes were studied and it was established the they were scattered over 16 chromosomes.
In the early 90s, André Goffeau organized about 30 labs who sequenced the entire yeast genome, over 6,200 genes. The sequences were published in 1996.
Many of these genes have now been cloned.

17. YEAST BIOTECHNOLOGY

18. CREATING BREWING YEAST THAT FERMENTS STARCH DIRECTLY
A wild yeast, Saccharomyces diastaticus, secretes glucoamylase, an enzyme that digests starch to form glucose.
We have cloned the STA gene that encodes glucoamylase and used the STA DNA to transform both lab strains and a brewing yeast strain so that they can ferment starch directly.

19. pSTA PLASMID

20. TRANSFORMING BREWING YEAST
TREAT CELLS WITH LiAc
ADD STA PLASMID DNA
PLATE ON HIGH COPPER MEDIUM
SELECT Cu-RESISTANT COLONIES
TEST FOR STARCH DIGESTION

21. TRANSFORMATION AND EXPRESSION OF STA GENE
PLASMID DNA IS REPLICATED AS A MINI-CHROMOSOME.
RNA POLYMERASE MAKES AN RNA COPY OF STA GENE.
RNA COPY IS TRANSLATED INTO GLUCOAMYLASE PROTEIN.
SECRETED ENZYME DIGESTS STARCH.
CELLS FERMENT STARCH!