Biological and Chemical Detection in the Brewing Industry

Submitted by:

David Jones

Brewing Process• Brewing bacteriology was born when

microorganisms responsible for the spoilage of beer were investigated by Louis Pasteur during the 19th 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.

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.

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)

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

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.

Automation PLC (Automationdirect)

Programmable Logic Controller

Automation PLC (Automationdirect)

Relay Out

Automation PLC (Automationdirect)Ladder

Programming View

Automation PLC (Automationdirect)

Drum Programming

Automation PLC (Automationdirect)

PID loops

Brewery Sensors

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

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

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

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

Glucose/Alcohol Electrode

• On-Line

• Response time < 2 Minutes

• Interoperable: yes small voltage, BNC connector

• Reproducibility ~ 3%

• Location: Mash, Fermentation, Packaging

Universal Sensors Inc.

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

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

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

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

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

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…

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

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.

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.

Conclusions

• Sensing and conditioning a signal is only half of the process.– Act and React: Interoperable with existing

systems• Overarching Control Scheme

• Manufacturing Procedures

Any Questions?