1. Biological and Chemical Detection in the Brewing Industry Submitted by: David Jones

2. Brewing Process Brewing bacteriology was born when microorganisms responsible for the spoilage of beer were investigated by Louis Pasteur during the 19 th century. He was called upon to determine why French beer was inferior to German Beer. Brewing has a long history; the process has been know for long time, but the science of why came later.

3. Brewing Process Malting – Barley is adjusted and kilned Milling – Exposes the starchy center Mash – Conversion of starch to sugar by alpha- amylase and beta-amylase Rinse/Boil – Kills bacteria and hops add bitter flavor Cooling/Fermentation – Yeast addition, conversion of fermentable sugars to carbon dioxide and alcohols.

4. Flavor Aspects Recipe Water Chemistry (Brewery Specific) Hops – strength and amount isomerized (a-acids) Yeast – Strain and Strength –Saccharomyces Ubarum –Saccharomyces Cerevisiae Fermentation: lag, log, rest, pH, dissolved oxygen, temperature, buffer capacity, carbon dioxide... Packaging – UV light, dissolved oxygen leads to bacterial spoilage (HACCP after fermentation) Flavor Agents: alcohols, sulfur compounds, esters, Di-acetyl and Pentane-2, 3-Dione, Polyphenols, Dimethylsulfide (DMS)

5. Fault Examples Off flavors: fruity, harsh, sweet, or bitter Haze: level of particles in suspension Lack of body – level of non-fermentable sugars and polyphenols Poor head retention or formation

6. Brewery Automation Complex chemical and biological process that needs to be controlled. Historically: Process controlled by manufacturing process (Brewery Specific) Today the manufacturing is driven by flexibility: The ability to produce a variety of beer using the same equipment.

7. Automation PLC (Automationdirect) Programmable Logic Controller

8. Automation PLC (Automationdirect) Relay Out

9. Automation PLC (Automationdirect) Ladder Programming View

10. Automation PLC (Automationdirect) Drum Programming

11. Automation PLC (Automationdirect) PID loops

12. Brewery Sensors

13. Dissolved Oxygen On Line Response time < 90 sec Interoperable: yes Immersion or Flow Through Performance: ± 1% of signal, max. ± 30 ppb Location: Fermentation Tank, HACCP Point after Boiling Stage Mettler Toledo

14. Carbon Dioxide On Line Response time <60 seconds for 90% step change Interoperable: Yes – Alarm relay output Non-dispersive infrared (NDIR) repeatable to ± 20 ppm 0-2000 ppm range Location: Fermentation ducts or HVAC ducts Veris Industries

15. Dissolved Carbon Dioxide At-Line Response time < 7 minutes Technology: fiber-optic, fluorescent dye Interoperable: Probable - 4-20 mA loop or 1-5 Vdc (analog output) Performance: ±5% of reading or 0.2% absolute Location: Fermentation Tanks YSI Life Science

16. pH no-Glass On-line Response time < 90 seconds Interoperable: yes – standard VP connection Performance: pH 0 ~14, 0 ~ 80°C Gel electrolyte, Argenthal electrode, Temperature Location: Mash tanks, Fermentation Tanks, Maturation, Packaging Mettler Toledo

17. Glucose/Alcohol Electrode On-Line Response time < 2 Minutes Interoperable: yes small voltage, BNC connector Reproducibility ~ 3% Location: Mash, Fermentation, Packaging Universal Sensors Inc.

18. Yeast Monitor At-Line Response time: Real-time Technology: Electrode, temperature, capacitance Interoperable: yes – RS232 port, alarm outputs 10 Cell size/strain positions Location: Fermentation, Maturation, Yeast Storage Aber Instruments

19. Yeast Monitor On-Line Response time: Real-time Technology: Radio frequency dielectrics and software –Measures Capacitance in living yeast cells (Plasma Membranes) Interoperable: yes – RS232 port and Ethernet, alarm outputs Location: Fermentation, Maturation, Yeast Storage Aber Instruments

20. Protein and Polyphenol Detection via Surface Plasma Resonance Off-line Response time: Rapid Confirmation Interoperable: No, standalone system Flexibility to determine multiple compounds with multiple sensor chip configurations Location: Brewing Lab (determine flavor constituents) Biacore

21. Gram Negative/ Gram Positive Bacteria Contamination Gram Negative –Acetic Acid Bacteria –Pectinatus cervisiiphilus –Enterobacteriaceae –Zymomonas –Pectinatus frisingensis –Selenomonas Lacticifex –Zymophilus raffinosivorans –Zymophilus paucivorans –Megaspaera Gram Positive –Lactobacillus –Lactic Acid bacteria –Pediococcus –Leuconostoc –Homofermentative cocci –Kocuria, Micrococcus and Staphylococcus –Endospore-forming bacteria

22. Biolog’s G+/G- Yeast detection Off-Line – Rapid detection Standalone System (the rest) MicroPlate ~ 96 different Chemical Substrates G+ >310; G- >500; Yeast >265 < 5 min Hand prep Photo-Optical (density) Automated database Location: Through-out

23. Biotrace’s ATP Bioluminescence All in one Test Kit: swabbing Off Line: Cleanliness Check ATP enzyme-driven light emissions Response time < 30 secs Measures light output (luminometer) or living cells Location: Throughout Specs:<150 RLU (pass) >300 RLU (Fail) Tanks, Valves, Doors …

24. Chemunex Chemscan RDI Off-line: Rapid Identification Technology: Combination of 1)Laser Scanning 2)cell labeling and 3)automated/database Fully Automated: 20 hr Presence; 1 hour direct Count High throughput ~ 60 samples per hour Drawback: lack of specific recognition of Brewery beer- spoilage bacteria. Subspecies of: –Pediococcus –Lactobacillus

25. Qualicon/DuPont’s RiboPrinting Off-Line: Rapid thorough detection ~8 hrs Technology: Random Amplified Polymorphic DNA PCR. Extracts: DNA; rRNA; compares gene sequence DNA extracted mixed with chemiluminescent; captured on digital image; Fingerprint is compared with database “RiboPrint” Powerful tool for identifying subspecies of beer-spoiling bacteria during the middle to late fermentation stages.

26. Conclusions Biosensor use is dependant upon many factors –Brewery size: Throughput, variety of brands, total energy used. –Cost: Potential Savings, Ability to integrate in to architecture, Installation cost, man power… –Potential Market: Better, Cheaper, Faster Anheuser-Busch: 47% of American Market, produced 127.9 Million barrels (101.8 M domestically), Gross sales of $15.686 billion dollars, and over 30 different brands. –Distributed Generation: The business of business is business. Distributed Manufacturing.

27. Conclusions Sensing and conditioning a signal is only half of the process. –Act and React: Interoperable with existing systems Overarching Control Scheme Manufacturing Procedures

28. Any Questions?