Background

Overview of Probiotic Raw Fermentation

Probiotics are a group of living microorganisms that, when ingested in sufficient quantities, can provide health benefits to the host. These microorganisms have positive health effects on the host mainly through symbiosis with the human body, competition or production of beneficial metabolites. Probiotics are commonly found in certain fermented foods, such as yogurt, sauerkraut, kefir, etc., and can also be taken in the form of supplements.

Probiotics mainly include a variety of lactic acid bacteria and some yeasts, of which lactic acid bacteria such as Lactobacillus acidophilus, Bifidobacterium and Lactobacillus plantarum, etc. It is the most common probiotic species and is widely used in fermented dairy products and probiotic supplements. Yeasts such as Saccharomyces boulardii, although fewer species, are also recognized as having probiotic properties. These probiotics provide a variety of benefits to human health by promoting intestinal health, enhancing immunity and other mechanisms.

Fig. 1. Pipeline for the assessment of probiotic safety. (Falzone, et al., 2024)

The benefits of probiotics include improving the balance of the gut microbiome, boosting host immunity, promoting digestion and nutrient absorption, helping to prevent and treat certain digestive diseases, and possibly having a positive effect on reducing allergic reactions and maintaining overall health. Although both prebiotics and probiotics are beneficial to gut health, they differ in nature and mechanism of action: Prebiotics are a class of non-digestible food ingredients that primarily serve as food for beneficial bacteria in the gut, promoting the growth and activity of these beneficial bacteria, while probiotics are living microorganisms that directly colonize the host gut and exert a range of positive health effects. In short, prebiotics are "food" for probiotics, which are living microbes that directly benefit the host.

By selecting suitable probiotic strains, optimizing fermentation conditions and controlling various parameters in the production process, Creative BioMart Microbe is committed to providing customers with customized probiotic fermentation service in food and producing satisfactory fermentation products. Our capacity to expand production seamlessly, starting from laboratory scales, through intermediate pilot phases, to full-scale manufacturing, allows us to provide our clients with diverse opportunities to bring innovative probiotic strains to both human and animal health sectors. Please feel free to contact us for more information.

Service Procedure

Service Details

Strain Screening

We have an extensive library of probiotic strains, all of which have been identified and tested for safety and assessed for quality. We will select the appropriate strain from the library based on your real needs, ensuring that the resulting product has specific health benefits and high activity.

The common probiotic strains we can offer include but are not limited to: Lactobacillus, Bifidobacterium, Streptococcus thermophilus, Saccharomyces boulardii, Saccharomyces cerevisiae, Bacillus subtilis and so on.

Scale-up Fermentation Production

We are equipped with world-class fermentation facilities and set up a dedicated fermentation project service team, with the ability and rich experience to provide small to large scale fermentation to achieve high production at low cost. During scale-up, computational fluid dynamics (CFD) simulations may be required to predict and optimize fluid behavior in large-scale bioreactors.

Fermentation Process Optimization

The production of probiotic raw materials involves a microbial fermentation process that requires the control of a variety of conditions, such as fermentation volume, stirring speed, ventilation, pH value to optimize the growth and metabolism of the strain. We have a variety of specifications of fermentation equipment with precise monitoring of the temperature and pH value.

Collection and Processing

Isolation of microbial cells from the medium by centrifugation or filtration techniques; Next, a washing step is used to remove the medium and other impurities to improve the purity of the probiotics. We offer lyophilization or spray drying to dehydrate probiotic cells to maintain their activity and facilitate storage and transportation; Finally, the dried probiotics are packaged in powder or other form.

Products Packaging and Storage

We set up multiple inspection points throughout the production process to ensure product quality and stability, including the monitoring and control of microorganisms, as well as the monitoring of the production environment. The final product is appropriately packaged to protect the probiotic activity. We ensure high quality packaging and offer a range of package sizes.

Our Advantages

• Extensive fermentation experience: We specialize in probiotic fermentation and have many years of industry experience.
• Flexible scale production: We have the ability to scale from laboratory scale to small pilot runs to mass production.
• High standard production systems: Manufacturing systems are designed according to high purity standards and industry best practices using cutting-edge fermentation equipment.
• Quality control: Work closely with the internal quality control laboratory to ensure the key steps of the production process and product quality.
• Customer services: We can focus on customer needs and provide customized contract fermentation services.

Case Study

Case Study 1: Optimal formula verification of strain 1 in a sample report.

Preliminary tests showed the original strain was infected with bacteria. We succeeded in getting the pure strain after three successive rounds of separation and purification. Optimal formulas were established by screening carbon, nitrogen sources and other factors. The inoculated processes of strain 1 to fermentation tank are studied and established. The OD value is significantly improved after the optimization of carbon and nitrogen sources. Compared with control group, the optimal group increases to 16 times. Preparation for 5 g dried biomass and 5 L fermentation supernatant of strain 1 was completed; the purity and activity are investigated and proved to be qualified.

Fig. 2. OD data of strain 1 before and after optimization.

Case Study 2: L. plantarum P-8 combined with S. thermophilus and L. delbrueckii subsp. bulgaricus, can be utilized to ferment milk.

In this research, the probiotic strain Lactobacillus plantarum P-8 was selected for the study. The impact of six different proportions (1:1, 1:5, 1:10, 1:50, 1:100, and 1:1000) of L. plantarum P-8 mixed with standard yogurt cultures was examined. Upon analysis, 66 distinct volatile substances, encompassing groups such as aldehydes, ketones, acids, alcohols, esters, and aromatics, were detected in the milk samples that had been fermented using these varying ratios on the day of production. Notably, at the 1:100 ratio, significant contributors to flavor, including 3-methylbutanal, hexanal, (E)-2-octenal, nonanal, 2-heptanone, 2-nonanone, and acetoin, were particularly prominent.

Fig. 3. Chromatographic fingerprints of all samples of milk fermented with different ratios of L. plantarum P-8. (Dan, et al., 2019)

Case Study 3: Lactobacillus acidophilus MTCC 10307 facilitates the conversion of sugarcane bagasse into phenolic flavor compounds.

This investigation explored the generation of beneficial phenolic flavor compounds through the use of Lactobacillus acidophilus MTCC 10307. Notably, ferulic acid, vanillic acid, and vanillin were identified as the key phenolic acids derived from the fermentation process. High-performance thin-layer chromatography was employed for the detection and quantification of these compounds, which were monitored over a 15-day period.

Ferulic acid reached its peak concentration on the 9th day of the incubation period, after which its levels began to diminish with continued incubation. In contrast, the highest levels of vanillic acid and vanillin were observed on the 12th day of incubation. On the 9th day, the amount of ferulic acid extracted per kilogram of sugarcane bagasse was approximately 275 mg, while on the 12th day, 18 mg and 15 mg of vanillic acid and vanillin were quantified per kg of bagasse.

Fig. 4. Release of phenolic acids from sugarcane bagasse by Lactobacillus acidophilus. (Pattnaik, et al., 2021)

FAQs

References:

1. Pattnaik B.; et al. Production of phenolic flavoring compounds from sugarcane bagasse by Lactobacillus acidophilus MTCC 10307. Arch Microbiol. 2021;204(1):23.
2. Dan T.; et al. Influence of Lactobacillus plantarum P-8 on Fermented Milk Flavor and Storage Stability. Front Microbiol. 2019;9:3133.
3. Falzone L.; et al. Benefits and concerns of probiotics: an overview of the potential genotoxicity of the colibactin-producing Escherichia coli Nissle 1917 strain. Gut Microbes. 2024;16(1):2397874.