Background

Microbial fermentation is a complex biological process that underpins the production of a vast array of products, from pharmaceuticals and biofuels to food ingredients and industrial chemicals. The success of fermentation processes, especially on an industrial scale, hinges on the integration of advanced engineering systems that are robust, reliable, and scalable. These systems form the backbone of the bioprocess, ensuring that the fermentation environment is meticulously controlled to optimize microbial growth and product yield. A satisfactory fermentation system must have the following characteristics:

Accurate Sensors: At the heart of these engineering systems are sensors that provide real-time data on various critical parameters. Accurate sensors are essential for monitoring factors such as pH, dissolved oxygen (DO), and temperature, which are vital for microbial metabolism. These sensors must be highly sensitive and precise to detect even minor fluctuations that could impact the fermentation process. They need to be calibrated regularly to maintain their accuracy and to provide reliable data for process control decisions.

Fast Response Time: The dynamic nature of microbial fermentation requires a system that can respond swiftly to changes in the bioreactor's conditions. A fast response time is crucial for implementing adjustments promptly, whether it's altering the agitation speed to enhance mixing, adjusting the airflow to maintain optimal DO levels, or modulating the temperature to prevent thermal stress on the microorganisms. This agility in response helps in maintaining the process within the desired set points and avoiding any potential downtime or loss in productivity.

Reliable Controls: The engineering systems must include reliable control mechanisms that can automatically adjust the bioreactor conditions based on the sensor inputs. These controls are typically managed by a sophisticated process control software that uses algorithms to make informed decisions. The reliability of these controls is paramount; they must operate without failure to ensure consistent process performance. Redundancy in control systems is often implemented to provide backup in case of a primary system failure, thereby ensuring uninterrupted operation.

Fig. 1. Soft sensor-based control set up for a generalised bioprocess. (Rathore, et al., 2021)

Scalable Engineering Systems: As fermentation processes move from laboratory to industrial scale, the engineering systems must be scalable to accommodate the increased volume without compromising on the precision of control. Scalability involves not only the physical size of the bioreactors and equipment but also the capacity of the control systems to manage larger volumes effectively. This includes the ability to maintain uniform mixing, appropriate nutrient distribution, and consistent environmental conditions across the entire volume of the bioreactor.

Integration and Automation: Modern fermentation facilities integrate these engineering systems with advanced automation technologies. Automation reduces human error, improves process reproducibility, and allows for the seamless collection and analysis of large datasets generated by the sensors. This integration enables a holistic view of the process, facilitating data-driven decision-making and process optimization.

Maintenance and Support: Robust engineering systems require regular maintenance to ensure they operate at peak performance. This includes routine checks, preventive maintenance, and prompt repair services. Support from the engineering system providers is crucial, especially for troubleshooting complex issues and providing technical expertise.

Service Procedure

Services Details

The fermentation process design services deal with novel bioreactor or biological process design, improved methods for recycling and separating fermentation products, improved bioreactor monitoring and control tools, and bioreactor performance optimization. Our services are not limited to general production, but also include methods to improve the production, extraction, purification and quantification of large molecules (such as enzymes, heterologous proteins, antibodies, lipids or RNA/DNA) or small molecules (such as alcohols, amino acids, carboxylic acids, aromatic compounds or sugars).

In addition, fermentation equipment design services mainly include the design, monitoring, biosensors and instrumentation, as well as downstream processing of fermented products, such as purification and extraction methods. The goal of these services is to improve production efficiency, product quality and process sustainability, while ensuring biosafety and biosecurity.

The Technical Fields Creative BioMart Microbe is Expert in:

• Continuous fermentation system
• Multi-effect differential pressure distillation
• Distillation system
• Evaporator design
• Neutral alcohol factory
• Bioethanol plant
• Cellulose raw material alcohol production plant
• Acetic acid production plant
• Glucose factory
• High fructose syrup factory
• Bakery yeast factory
• Citric acid factory
• Ethyl acetate production

As a senior microbial related technology research enterprise, Creative BioMart Microbe provides various types of fermentation engineering service packages to assist in your research project.

• Concept design

Our basic feasibility study can provide a basis for customers' investment decisions, including the determination of targets and factory performance, the selection and comparison of raw materials, final products and production capacity, detailed investigation of factory economic benefits, even the factory site selection and infrastructure design. More detailed data can cover product specifications, consumption value, cost assessment and project timetable.

• Basic engineering

Based on the needs of concept design, we will verify the design in the laboratory and provide the following documents: flow chart, material balance, process flow description, emission point list, terminal point list, operation manual, analysis method, equipment specifications and implementation standards, layout and basic plan, installation and acceptance criteria, timetable. We additionally offer performance testing services during plant construction and commissioning.

• Detailed engineering

In this stage, we will carry out the detailed design of pipelines, vessels and equipment, the automation of the plant and the integration of all systems. The delivered results include stress calculation, design drawings, three-dimensional graphics, welding layout, process control system, system programming and test guidance. All designs will be implemented according to the standards and rules of the customer's location.

• Purchase agreement

We coordinate the procurement process by providing technical or commercial assistance. The former concludes supplier list, quotation comparison and purchase recommendations, review and approval of key supplier specifications and detailed designs, manufacturing supervision, acceptance testing, and inspection documents. The later covers contract design, purchase terms, planned purchase behavior, packaging, and marking instructions.

• Supply

For ensuring the reliability of the process, we can provide customized equipment from key equipment components to complete automation systems in various fermentation stages such as distillation, rectification, dehydration, and evaporation. Key components include, but are not limited to, distillation tower trays, molecular sieve materials for dehydration devices, ventilation device.

• Supervise

We provide construction supervision services during the execution of the project. The service content includes supervising the construction of the factory from a technical point of view, complying with the provided design, pre-commissioning including supervision of equipment dry and hydraulic testing, adjustment and installation, on-site acceptance testing of the process control system. Otherwise, we can also provide operator training, assessment service if necessary.

• Technical support

After the factory is handed over, we will continue to serve you in order to ensure the best operation and practical experience in daily operations, and to find potential improvements.

Fig. 2. Engineering Service Packages in Creative BioMart Microbe.

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.

Case Study

Case Study 1: Novel gas supply system leveraging the shaking motion of an incubator for the aeration of shake flasks in aerobic microbial cultures.

Traditional shake flask cultivation, fundamental in bioprocess studies, often falls short in providing adequate oxygen and gas exchange, impacting the precise measurement of microbial growth and metabolic rates. Here, researchers present a groundbreaking gas supply device that capitalizes on the shaking motion of an incubator to ensure a steady airflow, effectively overcoming these challenges. They evaluated the system's performance by measuring the mass transfer coefficient (kLa) and growing Corynebacterium glutamicum H36LsGAD across different working volumes. Findings indicate that the novel gas supply system markedly surpasses standard silicone stoppers in oxygen provision, with kLa values reaching 2531.7 h^-1 as opposed to 20.25 h^-1 at a shaking speed of 230 rpm. Additionally, the new apparatus supported enhanced microbial growth in batch cultures, sustaining exponential growth rates even at higher working volumes.

Fig. 3. kLa values of flasks using gas supply apparatus (A) and silicone stopper (B). (Jung, et al., 2024)

Case Study 2: Gas supply apparatus with rotational motion of shaking incubator for the industrial production of glutathione.

In this study, scientists genetically modified Escherichia coli to optimize glutathione synthesis. The strain yielded 4.3 g/L of glutathione by boosting the expression of gshA and gshB, the genes responsible for the enzymes cysteine glutamate ligase and glutathione synthetase, with a majority of the glutathione being secreted into the surrounding culture liquid. The modified strain, KG06, generated a substantial 19.6 g/L of glutathione following a 48-hour fed-batch fermentation process, during which ammonium sulfate was persistently supplied as a sulfur source. The research also revealed that a steady supply of glycine was essential for enhancing glutathione output. Metabolic flux and metabolomic studies pinpointed the transformation of O-acetylserine into cysteine as a key constraint in KG06's glutathione synthesis. This bottleneck was significantly mitigated by employing sodium thiosulfate, which propelled the glutathione concentration to an unprecedented 22.0 g/L in the literature, as far as we are aware.

Fig. 4. KG06 was cultivated using sodium sulfate (A) or sodium thiosulfate (B) in a 5 L bioreactor. (Mori, et al., 2024)

Case Study 3: Modulating the production process of volatile fatty acids (VFA) from proteins shifts the dynamics of the bacterial community involved.

The focus of this case study is the emerging interest in producing VFA through mixed culture fermentation (MCF), where the emphasis has traditionally been on carbohydrate fermentation, with less exploration of proteins as substrates. This research investigates the impact of operational parameters: pH, protein source, and micronutrient supplementation—on the microbial makeup during the anaerobic digestion of protein-rich waste streams. Two continuous stirred tank reactors (CSTRs) were utilized, each fed with different proteins (casein and gelatin) and subjected to varying conditions: three pH levels (5.0, 7.0, and 9.0) with macronutrient addition, and two pH levels (5.0 and 7.0) with additional micronutrients. Throughout the study, Firmicutes, Proteobacteria, and Bacteroidetes were the predominant phyla, with their proportions fluctuating based on the operational variables. At neutral and alkaline pH levels, the type of protein predominantly influenced the microbial composition, whereas at pH 5.0, the acidity was the primary influence. The effect of micronutrient supplementation was observed to be pH and protein type-dependent, particularly affecting the populations of Clostridiales and Bacteroidales.

Fig. 5. Bacterial community composition at order level for casein (left) and gelatine (right) reactors at the different pH values. (Vijande, et al., 2024)

FAQs

References:

1. Rathore AS.; et al. Bioprocess Control: Current Progress and Future Perspectives. Life (Basel). 2021;11(6):557.
2. Jung M.; et al. Gas supply apparatus using rotational motion of shaking incubator for flask culture of aerobic microorganisms. Eng Life Sci. 2024;24(7):e2300243.
3. Mori H.; et al. Engineering Escherichia coli for efficient glutathione production. Metab Eng. 2024;84:180-190.
4. Vijande C.; et al. Altering operational conditions during protein fermentation to volatile fatty acids modifies the associated bacterial community. Microb Biotechnol. 2024;17(6):e14505.