Microbial cells cultivation is not only the origin, but also the foundation of microbiology. Researches in microbiology can only be carried out when the microbial cells can be cultured. However, conventional microbial cell cultivation is not only time consuming and labour intensive, but human error is also inevitable. Recent years, automated, modularised microbial cells micro-cultivation systems with small volume, good controllability, and equipped with real-time monitoring system have attracted great attention in microbiology. This review presents the state-of-the-art micro-cultivation systems which are implemented in microbial cells cultivation. The key development, applications of various system classified based on their construction, and the prospects of micro-cultivation system are discussed and insights into them are also provided.
Bioreactors ; Humans

Aims: Daddawa is a traditional fermented condiment produced from legumes in Nigeria. Lima bean is an underutilized legume in Nigeria. Natural fermentation has been the conventional method of producing daddawa but the product has been found to be of low quality and consistency. The present study aimed at comparing the microbial and biochemical changes during natural and controlled fermentation of lima bean for production of daddawa. Methodology and results: Lima bean was fermented into Daddawa naturally. It was also fermented into daddawa using pure starter culture of Bacillus subtilis and Bacillus pumilus as single starter. The microbial and biochemical changes during both fermentation conditions were evaluated. Lima bean fermented naturally (NF) recorded the highest total viable count at 48 h and 72 h of fermentation respectively. Alpha amylase and protease activities increased with fermentation, and reached their peak at 48 h in both naturally fermented lima bean and pure culture fermented lima bean samples. Lima bean fermented with B. subtilis (FBS) recorded the highest total free amino acids at 72 h (54.45 Glycine/g dry wt.). Conclusion, significance and impact of study: The use of lima bean for daddawa production enhanced its utilization. Controlled fermentation of lima bean by Bacillus species improved the biochemical properties such as α-amylase and protease activities and free amino acids content of fermenting lima beans into daddawa. Keywords: Daddawa; fermentation; lima bean; microbial; biochemical changes
Fermentation ; Bioreactors

Micro- and mini-bioreactors are characterized by their miniature working volume and comprehensive monitoring of process data, e.g., biomass, pH, dissolved oxygen, and fluorescence that are on par with conventional bench-top systems. The technical advancements of micro- and mini-bioreactors are supported by single-use material and micro-manufacturing, non-invasive optical sensors, automation such as industrial robotics and the integration of design of experiment software with data acquisition and process control. Owing to the miniature scales, micro-bioreactors typically feature lower turbulence intensity and energy dissipation rate, resulting in different mass transfer, mixing and shear conditions as compared to industrial scale equipment. Mini-bioreactors, nevertheless, are closer to large vessels. Micro- and mini-bioreactors are used mostly in screening and process development nowadays, owing to their combined high throughput and richness of data. They are also the hardware that will enable "precision medicine" in the near future.
Biomass ; Bioreactors ; Oxygen

The multiple correlation coefficient between the values determined by dry weight and those determined by fluorometer was observed with r = 0.96 and the standard error of calibration was 0.034. Using the best calibration data, in order to reconfirm the reliability of the fluorometer results in comparison with those obtained by dry weight on the cell concentration, fedbatch cultures were carried out. The results obtained by fluorometer measurements were in good agreement with those obtained by dry weight. The on-line monitoring of cell concentration by the fermentor system linked to a computer equipped with fluorometer was successfully carried out.
Bioreactors ; Calibration ; Pichia* ; Yeasts*

The effects of operational variables and reactor configurations (e.g. diameter of draft tube and the number of static mixers) on energy loss in modified airlift bioreactor were investigated at the first time. The results showed that improving the structure of draft tube could reduce energy loss in the bioreactor. When the diameter of draft tube and the number of static mixers were 4.0cm and 39, respectively, the total energy loss in the modified bioreactor was the least among all the configurations and 23.6% less than that of the conventional counterpart at the same air flowrate. The energy consumption for aeration was the smallest (43.9% less than that of the conventional counterpart) when the diameter of draft tube and the number of static mixers were 5.5cm and 13, respectively. The highest energy dissipation (70% - 80%) occurred in the riser, the bottom zone (about 20%) took the second place and the separator (less than 10%) took the third place. The energy dissipation in the downcomer was neglectable under the conditions in the research. When the energy loss per unit volume was considered, bottom zone stood the first place. It was implied that the riser was the most important zone to cut down the energy loss of the bioreactor and some attention should also be paid to the bottom zone.
Bioreactors ; Equipment Design

Currently, biomanufacturing technology and industry are receiving worldwide attention. However, there are still great challenges on bioprocess optimization and scale-up, including: lacing the process detection methods, which makes it difficult to meet the requirement of monitoring of key indicators and parameters; poor understanding of cell metabolism, which arouses problems to rationally achieve process optimization and regulation; the reactor environment is very different across the scales, resulting in low efficiency of stepwise scale-up. Considering the above key issues that need to be resolved, here we summarize the key technological innovations of the whole chain of fermentation process, i.e., real-time detection-dynamic regulation-rational scale-up, through case analysis. In the future, bioprocess design will be guided by a full lifecycle in-silico model integrating cellular physiology (spatiotemporal multiscale metabolic models) and fluid dynamics (CFD models). This will promote computer-aided design and development, accelerate the realization of large-scale intelligent production and serve to open a new era of green biomanufacturing.
Bioreactors ; Computer Simulation ; Fermentation ; Hydrodynamics

Industrial bioprocess is a complex systematic process and bio-manufacturing can be realized on the basis of understanding the metabolism process of living cells. In this article, the multi-scale optimization principle and practice of industrial fermentation process are reviewed, including multi-scale optimizing theory and equipment, on-line sensing technology for cellular macroscopic metabolism, and correlated analysis of physiological parameters. Furthermore, intelligent control of industrial bioprocess is further addressed, in terms of new sensing technology for intracellular physiological metabolism, big database establishment and data depth calculation, intelligent decision.
Bioreactors ; Biotechnology ; Fermentation ; Industrial Microbiology

With the rapid development of modern biotechnology, fermentation process is increasingly important in industrial production. To guarantee the stability of products, fermentation process should be elaborately monitored and controlled. Biomass is an important parameter for on-line monitoring in bioprocesses because biomass can reflect cell growth in a bioreactor directly. In-situ microscope, a non-invasive and image-analysis based technology, can real-time monitor cells in biological process. This review summarizes the development and application of in-situ microscopy in biomass monitoring.
Biomass ; Bioreactors ; Biotechnology ; Fermentation ; Microscopy

In this study, voltage was used as a disturbance factor to investigate the relationship between microbial community and methane (CH₄) production flux in a microbial electrolytic cell coupled anaerobic digestion (MEC-AD). Metabolic flux analysis (MFA) was used to explore the relationship between the CH₄ metabolic flux produced and the microbes. The results showed that both methane production flux and hydrogen production flux changed significantly upon voltage disturbance, while the voltage disturbance had little effect on acetic acid production flux. The maximum CH₄ production flux under 0.6 V disturbance was 0.522±0.051, which increased by 77% and 32%, respectively, compared with that of the control group under 1.0 V (0.295±0.013) and under 1.4 V (0.395±0.029). In addition, an average of 15.7%±2.9% of H₂ (flux) was used to reduce CO₂ to produce CH₄ and acetic acid, and an average of 27.7%±6.9% of acetic acid (flux) was converted to CH₄. Moreover, the abundance of Lachnospiraceae significantly affected the flux of acetic acid. The flux of CH₄ production is positively correlated with the abundances of Petrimonas, Syntrophomonas, Blvii28, and Acinetobacter, and negatively correlated with the abundances of Tuzzerella and Sphaerochaeta. The species that affected the flux of H₂ and CH₄ were similar, mostly belonging to Bacteroides, Clostridium, Pseudomonas and Firmicutes. Furthermore, the interspecies interaction is also an important factor affecting the MEC-AD methanogenesis flux.
Acetates ; Anaerobiosis ; Bioreactors ; Electrolysis ; Methane

Docosahexaenoic acid (DHA) has many unique physiological functions such as promoting the development of brain and retina in infants. Therefore, it is widely applied to food, pharmacy, breeding and other industries. To obtain engineered strains of Aurantiochytrium limacinum SR21 suitable for industrial application with increased lipid and DHA production, we designed a simple, fast, accurate and high-throughput screening method based on Nile red staining of oil droplets. First, ultraviolet C (UVC) mutagenesis was used to generate a random mutant library of A. limacinum. Second, screening conditions were optimized including staining conditions of Nile red and the sorting criterion. Thereby, three putative high-lipid mutants (D03432, D05106 and D01521) were selected from the mutant library containing 3 648 mutated clones. The three mutants grew faster and produced higher amounts of lipids and DHA compared to wild type (WT). In 100 mL cultures, the lipid content of D03432 and D05106 mutants reached 64.74% and 63.13% of dry cell weight respectively, whereas the wild strain exhibited only 43.19%. DHA yield in these two mutants were even 2.26-fold and 2.37-fold higher than that of the wild strain. Experiment with 5 L fermentor further confirmed that D03432 and D05106 mutants had better performance than the wild strain on DHA yield (45.51% and 66.46% more than that of the wild strain, respectively), and demonstrated their high potential for industrial application. This work not only generated several high-DHA content mutants with high potential for industrial use, but also provided vital guidance for high-throughput screening of lipid hyper-accumulating mutants in other oil-producing microorganisms.
Bioreactors ; Docosahexaenoic Acids ; Mutagenesis ; Staining and Labeling ; Stramenopiles