<< Return to Industry Reflections

2B or Not 2B? Success Criteria for Economical Bacillus Scale-Up

Typically, one of the first questions a contract fermentation customer will ask us is how quickly we can get them to economical commercial production. From our years of experience, we commonly find that if the customer can get 2 Billion CFU/ml (2B) in a standard media such as Trypticase Soy Broth (TSB) or Nutrient Broth (NB), we tend to be able to get them to commercial production in 8-12 weeks.

However, if the customer is in the 107 or 108 CFU/ml range in this quick test, we know we have a good amount of work ahead of us and it may take a while. Mind you, we have brought many strains with lower than “2B” CFU/ml yield to a point of economic feasibility, but you can bet it increases the production timeline quite a bit. Now, we know the last thing a customer wants to hear is that production yields may delay or even prevent commercialization all together. In order to prevent this, we highly recommend adding a simple 2B TSB screen into your earlier processes.

Planning Ahead

Right now, many scientists are trying to find the perfect strain for their application. They will spend months if not years screening their Bacillus libraries seeking desired attributes such as anti-fungal activity, plant growth promotion properties, hydrocarbon degradation or enzyme production properties. Once they find this wonder strain, the temptation is to immediately run to field trials or begin the arduous patenting process, both of which can take years to complete.

While you may be excited to run with your new proprietary strain, we urge you to take a precautionary step by performing an additional assay or 2B TSB screen when choosing between your library strains. We have seen many cases with customer strain candidates, where a scientist had invested 5-7 years of lab and fieldwork, only to find out in the end, economical production was not going to happen with us or other contract manufacturers. At the end of the day, some strains just do not want to scale up to some of these higher concentrations.

By adding this important test within your library strain selection processes, you could find that you may be better off choosing the strain with maybe 90-95% of the antifungal producing attribute compared to your best strain. That is if it also performs well during the 2B fermentation quick screen.

2B TSB Screen

When implementing this test, we would suggest taking a simple laboratory media such as Trypticase Soy Broth or Nutrient Broth and screen your top strains for the ability to achieve 2 Billion CFU/ml of spores in 48-72 hrs of aerobic growth (200-250 rpm shaking in a baffled flask). If you can achieve that number, you are in luck. You will likely move very quickly to an organism yielding 12-15 Billion CFU/ml with a chance of 20-30 Billion CFU/ml in the commercial fermenter. This will translate to 4×1011 to 2×1012 CFU/gram of freeze or spray dried powder in 8-12 weeks.

Before You Go to the Field, Check the Fermentation Yield

In summary, we know the pain of having invested so much time into your favorite strain only to find out that production yield will delay or prevent commercialization.  Once you have gone deep into field trials or patenting, it is so hard to turn back. Therefore, our advice is to add this screen way back during the library development stage. Do yourself a favor, take the extra couple of weeks to test your strains for economic viability. You will win in the end if you test for “2B or not 2B”.

About the Author:

Michael R. King, Ph.D., is CEO/CSO and the founder of Microbial Discovery Group (MDG). Michael is passionate about leading and developing dynamic teams in the Franklin and Oak Creek, WI facilities who share a common vision to Feed, Clean and Save the World.

Dr. King received both his Bachelor of Animal Science and Masters of Food Science Microbiology degrees from Oklahoma State University, followed by a PhD in Food Science Microbiology from the University of Illinois. Prior to founding Microbial Discovery Group, Michael held research and leadership positions within Nalco Chemical Company, Semco Bioscience and Agtech Products. With 25+ years of experience in microbial biosciences, Michael is committed to the continued exploration and advancement of biological solutions that leave the planet in a better place for generations to come.

Search Insights
Recently Posted Insights

The Commercialization of Microbial Solutions

As a Bacillus manufacturer, we work to grow our partners’ pure single-strain Bacillus cultures and scale the process so that they receive exactly what they need. Our success in the commercialization of microbial solutions comes from our trusted process and our...

How to Handle Odors Within Septic Systems After Spring Thaw

Spring’s warmer weather and longer days are always welcome after a cold winter, but for septic tank owners and operators, this seasonal transition can bring a significant problem: odors. Now that spring thaw has occurred and the heat of summer is setting in, septic...

In-Field and In-House Expertise in Wastewater Treatment

When your wastewater customers are facing challenges, you want a partner with proven success and extensive expertise in wastewater treatment. At Microbial Discovery Group, we can be that partner. MDG stands as a leading expert in bioaugmentation, holding in-field and...


Since the completion of our Oak Creek facility in the fall of 2022, we have steadily grown our capacities and capabilities as a trusted provider of Bacillus-based solutions. We are excited to announce that our growth has given us the opportunity to expand our...

Upcoming Webinar: Creating Quality Microbial Products

Producing high-quality microbial products for industrial, institutional, and consumer applications is essential for our partners. In our upcoming webinar, we will explore the critical aspects of microbial product development and how we ensure quality products for your...

Proven Success Treating Sludge with Biotifx®

If your customers are working in the biological wastewater treatment industry, chances are they’re dealing with the challenges of organic sludge. Sludge, the waste components left over after the wastewater has been treated, is a constant expense for facilities, and...

Sludge Tanks 101: Treating the Waste

Throughout the wastewater treatment process, certain components are left over that get separated from the treated water. This collection of leftovers is called sludge, which is then further treated in tanks. At Microbial Discovery Group (MDG), we have explored the...

Lagoons 101: Identification and Long-Term Treatment Approach

Lagoons and ponds have been used for treating wastewater for over 3,000 years. Today, there are over 8,000 wastewater treatment lagoons operating in the United States. One appeal to using lagoons is that they generally require less energy than other treatment systems...

Industry Reflections

Bacillus, Distinguishing Species and Strains Part I

Bacillus Species Identification Methods: Definitions, Pros, and Cons Understanding the effectiveness of Bacillus products is easy to understand because in many cases we can visibly see the difference. But, it is hard to see what exactly makes up the product. Like most companies, you have probably asked suppliers “what

Fats, Oils and Grease: Bacillus vs. The Alternatives

What do many commercial, residential, industrial and institutional facilities all have in common? If you said a door, you’re technically correct; but for the purpose of today’s article the correct answer was Fats, Oils and Grease (FOG)! All of your customers facilities have drains, and drains are sought-after hosts


What is bioaugmentation?

At MDG, we continue to discover all the possibilities to treat wastewater with Bacillus. Through years of research and development, we’ve found that bioaugmentation is the answer for wastewater treatment. Bioaugmentation is the process of adding scientifically selected organisms to a microbial community to enhance that community. The term bioaugmentation

Pulp and Paper Lagoons Problems

It’s been a tough few years for the pulp and paper (P&P) industry. With an increase in foreign competition and a lowered demand for paper, many facilities have had to tighten their belt. One of the most probable victims of these cuts are the wastewater treatment systems. Wastewater needs

Supplying Quality: One Strain at a Time

In the large-scale liquid bacterial fermentation world, there are two primary ways to grow bacteria: as individual strains or in combination with others. At Microbial Discovery Group, we grow one strain at a time as it has served us well over the past decade. Truth be told, if I

News and Insights

The Benefits of Liquid Dosing

Many wastewater distributors are faced with finding an efficient product to combat common challenges such as sludge, fats, oils, and grease, and odors. Bacillus-based products have been proven highly effective and reliable to reduce, eliminate, and alleviate these challenges in the wastewater market. Did You Know These Products Come in

Are You Offering a Bacillus RV Septic Treatment?

Outdoor activities like RV camping are an ideal way for travelers to escape indoor isolation. It’s all about packing up the necessities and hitting the open road. Living out of an RV camper for any amount of time will likely include eating, sleeping, and using the bathroom. These are

Top 5 Tips to Prevent Septic System Back-Ups

According to the EPA, about 1 in 5 US homes depend on septic systems, and with the uptick in professionals working from home, septic tank maintenance and backup prevention tips are among some of the top online searches. Septic service companies are in high demand, more people are home

FAQs on Bacillus Strains in the I&I Market

1. How do MDG’s Bacillus strains help reduce a variety of organic materials such as debris, decaying food, human and pet waste and FOG? MDG offers Bacillus-based products with different Bacillus strains that have a wide array of capabilities. Bacillus are living organisms and need food to live and multiply. They break down and consume