1. MYCOTOXINS
AND
BREWING TECHNOLOGY
BY
APEH, DANIEL O.
MTECH/SSSE/2011/2892
SUPERVISED BY
DR. MAKUN, HUSSAINI A.
DR.(MRS) MUHAMMAD, HADIZA L.
DEPARTMENT OF BIOCHEMISTRY
FEDERAL UNIVERSITY OF TECHNOLOGY, MINNA

2. INTRODUCTION
 Beer refers to beverages resulting from the germination and
fermentation of starches mainly derived from cereal (Gutcho, 1976).
 The third most popular drink overall after water and tea (en.wikipedia.org).
 Africa is the third largest beer consuming continent (WHO, 2004).
 Nigeria is among the first 30 top world consumers (FAO, 2003; WHO, 2004)
 High demand for beer across various continents equals high risk
of consumption of beer associated toxins (Karolína et al., 2012).
 Mycotoxins are major cereal contaminants (Reviewed in Makun, et al.,
2009; Chandrashekar et al., 2000).

 Processed cereals in form of beer may be contaminated with
mycotoxins.

3. MYCOTOXINS
 Over 300 fungi produced toxic compounds that
contaminate a wide variety of agricultural
commodities (Nielson and Smedsgaard, 2003)
 Produced as secondary metabolites (Pitt, 2000).
 At Pre-harvest, Post-harvest or During storage
(Reviewed in Kumar et al., 2008)
 Ingestion cause a range of toxic responses, from
acute to chronic health disorders (Roger, 1993)
 Affect trade
 Major producers
Aspergillus species
Penicillium species
Fusarium species

4. Aflatoxin B1 Aflatoxin B2 Aflatoxin G1 Aflatoxin G2 Aflatoxin M1
Zearalenone Fumonisin Patulin Citrulin Ergot Alkaloid
FIGURE 1: STRUCTURAL REPRESENTATION OF SOME MYCOTOXINS

5. HEALTH IMPACT OF MYCOTOXINS
 Diseases resulting from mycotoxin exposure are called
mycotoxicosis
They elicit short or long term impacts(Wayne, 2007)
 Inhibition
of metabolic pathways
 Impairment of growth and development
 Immunosupression
 Carcinogenicity, mutagenicity and teratogenicity
 T-2 toxin was responsible for the deaths of several
Russians from 1942-1947 (Gao and Yoshizawa, 1997).
 Aflatoxin intake relates to high incidence of liver
cancer is S.A, Kenya etc. (Fabio, 1999); Aflatoxicosis

6. ECONOMIC IMPACT OF MYCOTOXINS
 Mycotoxin contamination affects up to 25% of global
food and feed (FAO, 2009).
 Crop losses and Reduced animal productivity
 Costs from improving technologies for production,
storage and transport of crops
 Member states of the African Groundnut Council—The
Gambia, Mali, Niger, Nigeria, Senegal, and Sudan—have
calculated the annual cost of implementing a program to reduce
aflatoxin contamination at US$7.5 million (Atanda, 2011).
 The cost of analytical testing, especially as regulations
become more stringent
 EU regulation on aflatoxins cost Africa US$750 million each
year in exports of cereals, dried fruit and nuts (World bank
study; retrieved from http://www.mycotoxins.com)

7. BREWING TECHNOLOGY
 Refers to the processes involved
in beer production (Ertan Anli
and Mert, 2010)
 The most commonly used cereal
grain is barley; others are
wheat, maize (corn), sorghum
and rice (Gutcho, 1976).

8. Grain (Barley, Sorghum, Wheat) Moisture content
Microorganisms
Malting (3 stages) Gibberellic Acid
Hydrolytic Enzymes (Gutcho, 1976).
Adjunct
Malt
(Rice, Wheat,
Ground malt + Adjuncts
Maize, Barley Amylases and proteases hydrolysis & extraction
e.t.c) Mashing Increase in particle size (Milling)
Separation Wort is boiled sometimes with hops
Pre-treatment Boiling; Concentrate and sterilize the wort
Inactivate enzymes
Boiling Coagulate and precipitate protein
Hop
Caramelize the sugar slightly in order
Cooling to develop the color of the beer
Syrups Hops contribute antiseptic compounds
Sugars Wort
Yeast is inoculated in Wort
Fermentation Yeast converts sugar into organic compounds
Yeast C6H12O6+2Pi+2ADP 2C2H5OH+2CO2+ 2ATP
Glucose Ethanol + 2H2O
Maturation
BEER Lactic acid bacteria is inoculated in Wort
Results in souring (Lerio, 1993)
FIGURE 2: GENERAL CHART OF THE BREWING PROCESS

9. MYCOTOXIN AND BREWING
 Some mycotoxins seems to survive major beer
production processes namely malting,
mashing, boiling and fermentation into beer
(Scott, 1996)
 Mycotoxins in beer could either be developed
or detoxified at various brewing processes
(Mably et al., 2005)

10. Grain (Sorghum, Wheat) Steeping resulted in about 45%
Malting AFL lost (Yahl, 1971; Romer, 1984)
Malt
Adjunct Adjunct is a source of AFL
(Rice, Wheat, Maize, B Mashing
arley e.t.c)
contamination (Karolína et al., 2012)
Separation
Pre-treatment
At 100°C and 250°C for 30min
Boiling showed 10.4% and 99% reduction
Hop
in AFB1 (Oluwafemi and Ikeowa, 2005)
Cooling
Syrups
Sugars Wort For 72hrs showed 50% reduction
of AFB1 (Yuan et al., 2008)
Yeast Fermentation
Lactic acid fermentations lead to
Maturation opening of the lactone ring (Nout 1994).
BEER
Upto 89% AFB1 was removed during Lager beer processing (Oluwafemi, 2004)
FIGURE 3: AFLATOXIN AND BREWING

11. Grain (Sorghum, Wheat)
Almost completely lost
Malting ( Krogh et al., 1974).
Malt
Adjunct
(Rice, Wheat, Maize, B Mashing 40–89% was lost (Sylvie et
arley e.t.c) al., 2011)
Separation
Pre-treatment
Boiling Most probably due to
Hop proteolytic degradation
Cooling (Ertan Mert, 2010).
Syrups
Wort
Sugars
Fermentation Destroyed in the fermentation
Yeast
process (Kostecki et al., 1991;
Maturation
SCOOP, 2010)
BEER
Higher in the ‘‘non-alcoholic’’ beer (Tangni and Lanrondelle, 2003)
Upto 70% of the OTA was degraded in the production steps beer(Nip et al., 1975
FIGURE 4: OCHRATOXIN AND BREWING

12. Grain (Sorghum, Wheat) Increase in Fusarium
Malting spp, and fumonisin
Malt
Brewing adjuncts e.g
Adjunct (Rice, Wheat,
Maize, Barley e.t.c)
Mashing corn contributes fumonisin
(Scott and Lawrence, 1995).
Separation
Pre-treatment
Boiling
Hop Heat-stable (Alberts et al.,
1990)
Cooling
Syrups
Wort Level change not
Sugars
Fermentation significant (Scott et al., 1993).
Yeast
Maturation
BEER
Levels above maximum limit has been reported in severel countries
(Drager, 1996; Mbugua and Gathumbi) e.g 150±24 ng/ml in Cameroonian beer
(Roger, 2011).
Gushing increases with fumonisin in beer (Casey,1996;).
FIGURE 5: FUMONISIN AND BREWING

13. Grain (Sorghum, Wheat) Increased by 18–114% of
Malting that present on the
Malt
original barley in 5 day
Adjunct
malts, (Scott, 1996; Schwarz et
(Rice, Wheat, Maize, Ba
Mashing al., 2005, Lancova et al., 2008)
rley e.t.c)
Separation
Pre-treatment
Boiling Stable after 7-9 days of
Hop alcoholic fermentation with S.
Cooling
Syrups
cerevisiae (Scott, 1992)
Wort
Detected at high levels in both
Sugars Fermentation the solid residue and the
Yeast fermented liquid (Bennet and
Maturation
Richard 1996).
BEER
Trichothecenes are stable to brewing process; occurring in commercial
beer in many countries (Shim et al., 1997; Molto et al., 2000; Baxter et al., 2006 ).
After brewing, 80–93% of DON present on the malt grist was detected in
the beer (Schwarz et al., 2005).
FIGURE 6: DEOXYNIVALENOL (DON) AND BREWING

14. TABLE 2 : YEAST AND/OR YEAST CELL WALL MATERIAL AS
POTENTIAL MYCOTOXIN DECONTAMINATING AGENTS
AGENT Mycotoxin bound Level of Binding Reference
S. cerevisiae AflatoxinB1 > 60% Shetty and Jespersen,
2006; Devegowda et
al., 1996
Yeast from West AflatoxinB1 >60% Shetty and Jespersen,
African maize 2006
Cansida crusei AflatoxinB1 15- 39% Shetty and Jespersen,
2006
Mannan- Ochratoxins & High binding Raju & Devegowda,
oligosaccharides Zearalenone 2000; Devegowda et
DON&FumonisinB1 Little binding al., 1996
Modified mannan- AflatoxinB1 77% Devegowda et al., 1996
oligosaccharides from
the S. cerevisiae
Mixture of 40% (w/w) Ochratoxin A High binding Grunkemeier, 1990
sterilized yeast + 60%
(w/w) residue of beer
fermentation

15. TABLE 3 : LACTIC ACID BACTERIA (LAB) AND/OR LAB MATERIAL
AS POTENTIAL MYCOTOXIN DECONTAMINATING AGENTS
AGENT Mycotoxin bound Level of Reference
Binding
Lactobacillus Aflatoxin B1 40-59% Shetty and
plantarum Jespersen, 2006
Paralactobacillus Aflatoxin B1 <15% Shetty and
serangolensis Jespersen, 2006
Lactobacillus Aflatoxin B1 (more than B2, 80% El–Nezami et al.,
rhamnosus G1, G2) 1998a
Lactobacillus Deoxynivalenol, 3- Effective El–Nezami,
rhamnosus strains acetyldeoxynivalenol, binding Chrevatidis,
LGG and LC 705 nivalenol, fusarenon, Auriola, Salminen,
diacetoxyscirpenol, T-2 toxin, & Mykkanen, 2002
and HT-2
Lactic Acid Bacteria Aflatoxins Inhibition (Coallier-Ascah &
of bio- Idziak, 1985;
synthesis Thyagaraja &
Hosono, 1994).

16. Retrieved from www.bioimages.net
FIGURE 7: SACCHAROMYCES CEREVISIAE (YEAST)
CELL WALL COMPONENTS

17. Retrieved from www.bioimages.net
FIGURE 8: LACTIC ACID BACTERIA CELL WALL COMPONENTS

18. BIOCHEMICAL
MECHANISMS
OF ACTION
OF SOME
MYCOTOXINS
CONTAMINATING
BEER

19. Dihydrosphingosine N-acyl
transferase is one cellular
target for fumonisin toxicity and
carcinogenicity
Inhibition of sphingolipid
biosynthesis (Wang et al., 1991).
Increases the ratio of sphingoid
precursors
Cell deregulation
Cell death (Riley et al., 1996)
TOXICOLOGY
 Suspected human carcinogen.
Toxic to pigs and poultry.
Cause equine eucoencepha-
lomalacia (ELEM), a fatal disease
of horses
FIGURE 11: MECHANISM OF ACTION OF FUMONISIN

20. DON inhibits Translation by
1. Interaction with the peptidyl transferase
centre on the 605 ribosomal subunit.
2. They also prevent release of the
polypeptide, by suppressing hydrolysis of
the peptidyl—tRNA at termination
 TOXICOLOGY
 Feed refusal
 Reduced weight gain
 Diarrhea
 Vomiting
FIGURE 12: MECHANISM OF ACTION OF DEOXYNIVALENOL

22. •Disruption Of Phenylalanine
Metabolism
•Reduced PEPCK at
Translation level
•Reduced Glyconeogenesis
•Cell Death
FIGURE 10: MECHANISM OF ACTION OF OCHRATOXIN

23. PREVENTION AND MANAGEMENT OF MYCOTOXIN
CONTAMINATION IN COMMODITIES
 Implementation of GAPs from planting to harvesting
 Proper storage conditions
 Biotechnology; Resistant crop varieties (Hell et al., 2011)
 Coordination between the different stakeholders
 The establishment of food safety measures needs a
legal basis, otherwise no enforcement is possible
 Public awareness/ University curriculum
 HACCP and others

24. REGULATIONS
 Established in about 100 countries of which 15 are
African.
 Agencies involved; WHO, FAO, WHO-JECFA,
EFSA, FSA, SON, ACADEMIC INSTITUTIONS
 Maximum allowable limits
4 µg/kg aflatoxinB1 for All foods (Nigeria standard)
 0.5 µg/kg DON for grain (USA standard)
 2-4 µg/kg FumunisinB1 for maize products (EU
standard)
5 µg/kg and 3 µg/kg Ochratoxin in cereals and
malt respectively (EU standard)

25. CONCLUSION
 Brewing technology is a multi-phased process, each
phase contributes differently to the final quantity of
different mycotoxins that may be found in beer.
 Grain and/or adjuncts may be source of contamination
by fungi and their mycotoxins.
 Malting increases the risk of fungi and mycotoxin
contamination but reduce ochratoxin levels.
 Fermentums binds mycotoxins on their cell walls.
 Generally, DON and other tricothecenes are shown to
increase across the brewing process, fumonisin is
relatively stable while aflatoxin and ochratoxin are
generally observed to be reduced across the brewing
process