The use of Schizosaccharomyces yeast in order to reduce the content of Biogenic Amines and Ethyl Carbamate in wines

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1 August 18, 2015 The use of Schizosaccharomyces yeast in order to reduce the content of Biogenic Amines and Ethyl Carbamate in wines Dept. Chemistry and Food Technology IS Prof. Santiago Benito Sáez.

2 Lecture objectives To give a summary about red wine and Food Safety => Main problems possible industrial solutions. To propose a specific alternative in order to manage two specific wine /Food Safety emerging problems: Biogenic amines. Ethyl Carbamate..

3 Introduction Wine => Easy to manage from Food Safety point of view.

4 Introduction London 1854 Broad Street (Cholera outbreak) What would you prefer to drink?

5 Today everything is different There are several problems related to wine - food safety (No high risk microorganisms => but other significant risks) Alcoholism S2 or other preservatives Biogenic Amines chratoxin A Physical hazards Aditives (Clarification) Ethyl Carbamate

6 Summary Main Wine Food Safety problems <=> Solutions Problem Ethanol Risk group Alcoholics, diabetics, Solution (HACCP)

7 General problems related to alcohol-alcoholism Most alcoholics do not drink wine (Spanish Society of Anonymous Alcoholics)

8 General problems related to alcohol Most alcoholics do not drink wine (Spanish Society of Anonymous Alcoholics) Most wine consumers=> Considered as responsible =>Things Alcoho

9 General problems related to alcohol Most alcoholics do not drink wine (Spanish Society of Anonymous Alcoholics) Most traffic accidents related to alcohol consumption do not depend on wine (Spanish Traffic Agency)

10 Problem Ethanol Risk group Alcoholics, dia eti s. Solution (HACCP) Labeled Free alcohol wine Responsible Consumers

11 Problem Ethanol Physical hazards Risk group Alcoholics, diabetics, Anyone Solution (HACCP) Labeled Free alcohol wine

12 Problem Physical hazards Risk group Solution (HACCP) Anyone Filtration

13 Problem Ethanol Physical hazards S2 Risk group Alcoholics, diabetics, Solution (HACCP) Labeled Free alcohol wine Anyone Filtration Asthmatics

14 Problem Risk group Solution (HACCP) Labeled: It contains sulfites. ther preservatives (sulfites free) S2 Asthmatics Legal limits Use other new preservatives or technologies

15 Problem Ethanol Physical hazards S2 Allergenic additives Risk group Alcoholics, diabetics, Solution (HACCP) Labeled Free alcohol wine Anyone Filtration Labeled: It contains sulfites. ther preservatives (sulfites free) Asthmatics allergic people (Ex: albumin) Legal limits

16 Problem Risk group Allergenic additives allergic people (Ex: albumin) Solution (HACCP) Labeled. r non allergenic additives Use of new additives or technologies

17 Problem Ethanol Physical hazards Risk group Alcoholics, diabetics, Solution (HACCP) Labeled Free alcohol wine Anyone Filtration Labeled: It contains sulfites. ther preservatives (sulfites free) S2 Asthmatics Allergenic allergic people additives (Ex: albumin) Biogenic allergic people Amines (Ex: Histamine) Legal limits Labeled. r non allergenic additives

18 Biogenic Amines Problem rigin thers: Decarboxylase enzymes -Tyramine -Putrescine -Cadaverine Histidine Histamine -Phenylethylamine Lactic Bacteria (. eni ) metabolism => Biogenic Amines

19 Biogenic Amines Problem rigin thers: Decarboxylase enzymes -Tyramine -Putrescine -Cadaverine Histidine Histamine -Phenylethylamine Lactic Bacteria (. eni ) metabolism => Biogenic Amines Real Group of Risk Alergic People => Biogenic Amines + Alcohol

20 Biogenic Amines Problem rigin thers: Decarboxylase enzymes -Tyramine -Putrescine -Cadaverine Histidine Histamine -Phenylethylamine Lactic Bacteria (. eni ) metabolism => Biogenic Amines Real Group of Risk Alergic People => Biogenic Amines + Alcohol Legal Limits Germany, Switzerland, Belgium, Austria, Holland, France => 2-10 mg/l (Histamine)

21 Biogenic Amines Problem rigin thers: Decarboxylase enzymes -Tyramine -Putrescine -Cadaverine Histamine Histidine -Phenylethylamine Lactic Bacteria (. eni ) metabolism => Biogenic Amines Real Group of Risk Alergic People => Biogenic Amines + Alcohol Legal Limits Germany, Switzerland, Belgium, Austria, Holland, France => 2-10 mg/l (Histamine) How should we control it?

22 Problem Ethanol Physical hazards Risk group Alcoholics, diabetics, Solution (HACCP) Labeled Free alcohol wine Anyone Filtration Labeled: It contains sulfites. ther preservatives (sulfites free) Asthmatics S2 Allergenic allergic people additives (Ex: albumin) Biogenic allergic people Amines (Ex: Histamine) Anyone Ethyl (carcinogenic) Carbamate Legal limits Labeled. r non allergenic additives legal limits => but no viable industrial solution?

23 Ethyl Carbamate Problem HN rigin H NH Arginine Arginase (Saccharomyces) H2 H2N Urea + H+ H2N Time + Ethanol Ethyl carbamate H Yeast metabolism => Ethyl Carbamate Precursor (UREA) Uthurry et al.2004; Bertrand 1993

24 Ethyl Carbamate Problem HN rigin H NH Arginine Arginase (Saccharomyces) H2N H2 Urea + H+ H2N Time + Ethanol Ethyl carbamate H Yeast metabolism => Ethyl Carbamate Precursor (UREA) Uthurry et al.2004; Bertrand 1993 HN H Citrulline H N H H NH Arginine deiminase transcarbamilase kinase rnithine H2N + Ethyl carbamate Lactic Bacteria (. eni ) metabolism => Ethyl Carbamate Tegmo-Larsson et al.1989

25 Ethyl Carbamate Problem HN rigin H NH Arginine Arginase (Saccharomyces) H2N H2 Urea + H+ H2N Time + Ethanol Ethyl carbamate H Yeast metabolism => Ethyl Carbamate Precursor (UREA) Uthurry et al.2004; Bertrand 1993 HN H Citrulline H N H H NH Arginine deiminase transcarbamilase kinase rnithine H2N + Ethyl carbamate Lactic Bacteria (. eni ) metabolism => Ethyl Carbamate Tegmo-Larsson et al.1989 Legal Limits Canada, USA (recomendation) and Japan => µg/l

26 Ethyl Carbamate Industrial Situation rigin Yeast metabolism => Ethyl Carbamate Precursor (UREA) Uthurry et al.2004; Bertrand 1993 Lactic Bacteria (. eni ) metabolism => Ethyl Carbamate Tegmo-Larsson et al.1989 Legal Limits Canada, USA (recomendation) and Japan => µg/l How should we control it?

27 Problem Ethanol Physical hazards Risk group Alcoholics, diabetics, Solution (HACCP) Labeled Free alcohol wine Anyone Filtration Labeled: It contains sulfites. ther preservatives (sulfites free) Asthmatics S2 Allergenic allergic people additives (Ex: albumin) Biogenic allergic people Amines (Ex: Histamine) Anyone Ethyl (carcinogenic) Carbamate Legal limits Labeled. r non allergenic additives legal limits => but no viable industrial solution? legal limits => but no viable industrial solution?

28 PRDUCTIN F RED WINE Reception Destemering Chrusering Prefermentative Maceration 1º Fermentation Maceration Pressing 2ª Fermentation Maceration Filtration. Stabilization Bottling Ageing

29 MICRBILGICAL SIMPLIFICATIN PRDUCTIN F RED WINE Reception Destemering Chrusering Red Grape Juice Prefermentative Maceration 1º Fermentation Maceration Pressing SUGARS ALCHL Yeast: S. cerevisiae 1º Fermentation 2ª Fermentation Maceration Filtration. Stabilization Bottling MALIC ACID LACTIC ACID Lactic Bacteria:. oeni 2º Fermentation Ageing Stabilized wine

30 INDUSTRIAL ALTERNATIVE PRPSAL MICRBILGICAL SIMPLIFICATIN Red Grape Juice SUGARS Red Grape Juice ALCHL MALIC ACID ETHANL SUGARS ALCHL Yeast: Schizo. pombe Yeast: S. cerevisiae nly 1º Fermentation 1º Fermentation MALIC ACID LACTIC ACID Stabilized wine Lactic Bacteria:. oeni We avoid possible collateral effects related to 2º Fermentation by Lactic Bacteria: -Biogenic amines -Ethyl carbamate 2º Fermentation Stabilized wine

31 Why to use Schizosaccharomyces selected strains? Classic use => deacidification (Malic Acid ) Recommended Practice by International rganization of Vine and Wine. Alternative to MaloLactic fermentation by Bacteria (Collateral effects ). Benito et al 2012; Benito et al 2014; Benito et al 2015 Urease Activity => Urea (Main Ethyl Carbamate precursor ) Lubbers et al. 1996; Déak.2008; Benito et al 2014 To avoid Biogenic Amines and E. Carbamate?

32 Classical Fermentations <=> Schizo Fermentations S. cerevisiae +. oeni Selected S.pombe Histamine Urea Histamina (mg/l) Benito et al Food Microbiology. 42: Benito et al Molecules. 20: Lower levels of Biogenic Amines and Urea in fermentations involving selected strains of S.pombe.

33 Conclusions Most wine/food Safety problems have a relatively easy solution. Using Schizosaccharomyces pombe fermentation technology is possible to control two specific wine/food Safety problems that are more complex: Biogenic amines. => Specific consumers Ethyl Carbamate. => Specific markets

34 Bibliography Benito S, Palomero P, Morata A, Calderón F, Suárez-Lépe JA (2012) New applications for Schizosaccharomyces pombe in the alcoholic fermentation of red wines. Int J Food Sci Tech 47: Benito S, Palomero F, Morata A, Calderon F, Palmero D, Suárez-Lepe JA (2013) Physiological features of Schizosaccharomyces pombe of interest in making of white wines. Eur Food Res Technol 236:29-36 Benito et al. (2013). Schizosaccharomyces selective differential media. Afr. J. Microbiol. Res. 7 (24), Benito S, Palomero P, Calderón F, Palmero D, Suárez-Lépe JA (2014a) Selection of Appropriate Schizosaccharomyces strains for winemaking. Food Microbiol 42: Benito S, Palomero P, Calderón F, Palmero D, Suárez-Lepe JA (2014b) Schizosaccharomyces. In: Batt CA, Tortorello ML (eds) Encyclopedia of Food Microbiology, vol 3, 2nd edn. Elsevier Ltd, Academic Press, Amsterdam, pp Benito S, Palomero P, Gálvez L, Morata A, Calderón F, Palmero D, Suárez-Lepe JA (2014c) Quality and Composition of Red Wine Fermented with Schizosaccharomyces pombe as Sole Fermentative Yeast, and in Mixed and Sequential Fermentations with Saccharomyces cerevisiae. Food Technol Biotechnol 52: Benito A, Palomero F, Calderón F, Benito S (2015) Combine Use of Selected Schizosaccharomyces pombe andlachancea thermotolerans Yeast Strains as an Alternative to thetraditional Malolactic Fermentation in Red Wine Production. Molecules 20:

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