Background

Fermentation technologies encompass a broad field and is the intentional use of fermentation by microorganisms such as bacteria and fungi as well as eukaryotic cells to make products useful to humans. Modern large-scale fermentation technologies have become highly automated and precisely controlled, enabling real-time monitoring and adjustment of key parameters within bioreactors such as temperature, pH, dissolved oxygen, etc. Process scaling techniques have evolved multiple strategies to overcome challenges from laboratory to industrial scale, including the use of statistical models and computational fluid dynamics simulations. As a sustainable production method, large-scale fermentation helps to reduce the dependence on fossil fuels and reduce the carbon footprint of the production process. Despite significant progress, large-scale fermentation still faces technical, economic, and regulatory challenges, including uncertainty about process scale-up, control of production costs, and regulatory compliance in emerging markets.

Fig. 1. Measurements during strain development in microtiter well plates, shake-flasks, and bioreactors. (Wehrs, et al., 2019)

Industrial microbial manufacturing, or the use of microbial fermentation to produce chemicals and materials, is playing an increasingly important role in chemistry and manufacturing. To that end, Creative BioMart Microbe has shown early scale-up to help ensure the target strain is commercially viable. We have a unique set of skills and tools that can be used to develop bio-manufacturing processes from microliters to 100 liters. Creative BioMart Microbe's ability to engineer and model biological systems has opened the way for the production of new and unnatural products.

Service Procedure

Platforms

Services Details

Our industrial microbial fermentation system with liquid handling robots and an integrated board reader platform allows us to perform high-throughput screening of up to hundreds of samples per day. This allows evaluation of a large number of variables, from process conditions to moderate components, using appropriate experimental designs.

We have six 500 ml bioreactors and has a large number of pilot scale reactors, equipped with a mass flow controller and the process control system, pH sensitive probe and DO probe, can monitor 5000 liters of fermenter progress, in order to realize accurate control, actively to further develop and optimize the existing process and development of new large-scale fermentation process.

During Large-scale process,Creative BioMart Microbe are trying to build industrial size fermenter capable or close of producing the fermentation products as efficient as those produced in small scale fermenters.

Our Advantages

• Full Staffing

• Have researchers from the field of fermentation engineering and microbiology, who can develop and optimize the whole fermentation process
• Have operators engaged in fermentation industry for many years, familiar with the operation of fermentation tank and various analytical equipment
• Have a professional QC team, able to abide by the FDA terms and conditions and other documents and technical documentation
• Have a timely technical support and customer service team, can real-time technical communication with customers, tracking services

• High Quality Fermentation Equipment

• Large scale fermenter, pilot scale fermenter and experimental fermenters
• Hollow fiber filtration system
• Frozen centrifuge

• A Variety of Expression Systems

We have a variety of expression systems, such as bacteria and fungi as well as eukaryotic cells like CHO cells and insect cells.

• Many Years of Experience in Large Scale Fermentation

We improve the large-scale fermentation tech to maintenance of constant power consumption per unit of broth and the maintenance of constant volumetric transfer rate.

Case Study

Case Study 1: 15 L trial fermentation optimization of strain 1 is completed and scale-up process of strain 1 is established.

The aim of this project is to obtain 5 L trial fermentation for strain 1. Optimal formulas were established by screening carbon, nitrogen sources and other factors. The inoculated processes of the strain to fermentation tank are studied and established. Preparation for 5 g dried biomass and 5 L fermentation supernatant of all three strains are completed; their purity and activity are investigated and proved to be qualified. After optimization, the quality of the seed is significantly improved. Feeding process has been developed in this process. The cultivate time is prolonged to 64 h at the point where the strain stops growing.

Fig. 2. Optimization of fermentation process of strain 1 in 15 L tank.

Case Study 2: Optimizing bioreactor conditions for Spirulina fermentation by Lactobacillus helveticus and Kluyveromyces marxianus.

This study focused on optimizing the production of fermented Spirulina (FS) products using a bioactivity-guided strategy with Lactobacillus helveticus B-4526 and Kluyveromyces marxianus Y-329 in a 3-L bioreactor. Various operating conditions, including aeration rates and pH modes, were tested. Screening revealed that "cascade" FS significantly decreased viability of colon cancer cells (HT-29) in a dose-dependent manner, with up to a 72 % reduction. Concentrations of "cascade" FS up to 500 μg/mL were found to be both safe and efficient in inhibiting the release of nitric oxide (NO), while maintaining cell viability. Moreover, "cascade" FS was also found to contain a variety of volatile organic compounds and helped in diminishing the typical "seaweed" scent.

Fig. 3. The growth of (a) L. helveticus, (b) K. marxianus. (Yay, et al., 2024)

Case Study 3: Fermentation process optimization of probiotic Bacillus coagulans, generating yields of (7.8 ± 0.2) × 10⁹ CFU/mL in 30-L fermenter.

Bacillus coagulans is a probiotic agent widely used in various industries. In this study, researchers isolated a novel strain of B. coagulans, X26, from soil and characterized its properties. To optimize the fermentation process of this bacterium, researchers adopted the response surface design. The yield of X26 in a 500-mL flask culture was (5.12 ± 0.01) × 10⁹ CFU/mL, and in a 30-L fermenter was (5.11 ± 0.02) × 10⁹ CFU/mL, accounting for a 9.9-fold higher field than that with a basal medium before optimization. Further, the fermentation process in the 30-L and a 10-T fermenter was optimized, generating yields of (7.8 ± 0.2) × 10⁹ CFU/mL (spore rate: 96.54%) and (8.7 ± 0.1) × 10⁹ CFU/mL (spore rate: 97.93%), respectively.

Fig. 4. 30-L fermenter process without feeding. (Xu, et al., 2023)

FAQs

References:

1. Wehrs M.; et al. Engineering Robust Production Microbes for Large-Scale Cultivation. Trends Microbiol. 2019;27(6):524-537.
2. Yay C.; et al. Optimizing bioreactor conditions for Spirulina fermentation by Lactobacillus helveticus and Kluyveromyces marxianus: Impact on chemical & bioactive properties. Bioresour Technol. 2024;403:130832.
3. Xu L.; et al. Isolation, functional evaluation, and fermentation process optimization of probiotic Bacillus coagulans. PLoS One. 2023;18(11):e0286944.