Practical Fermentation Process Research Articles

Distributionally robust parameter identification of a time-delay dynamical system with stochastic measurements

• A distributionally robust parameter identification model is proposed for time-delay systems. • The problem is transformed to a single-level optimization problem. • A gradient-based optimization method is developed to solve the transformed problem. In this paper, we consider a parameter identification problem involving a time-delay dynamical system, in which the measured data are stochastic variable . However, the probability distribution of this stochastic variable is not available and the only information we have is its first moment. This problem is formulated as a distributionally robust parameter identification problem governed by a time-delay dynamical system. Using duality theory of linear optimization in a probability space, the distributionally robust parameter identification problem, which is a bi-level optimization problem, is transformed into a single-level optimization problem with a semi-infinite constraint. By applying problem transformation and smoothing techniques, the semi-infinite constraint is approximated by a smooth constraint and the convergence of the smooth approximation method is established. Then, the gradients of the cost and constraint functions with respect to time-delay and parameters are derived. On this basis, a gradient-based optimization method for solving the transformed problem is developed. Finally, we present an example, arising in practical fermentation process, to illustrate the applicability of the proposed method.

Study on feasibility of determination of glucosamine content of fermentation process using a micro NIR spectrometer

N-acetyl-d-glucosamine (GlcNAc) is a microbial fermentation product, and NIR spectroscopy is an effective process analytical technology (PAT) tool in detecting the key quality attribute: the GlcNAc content. Meanwhile, the design of NIR spectrometers is under the trend of miniaturization, portability and low-cost nowadays. The aim of this study was to explore a portable micro NIR spectrometer with the fermentation process. First, FT-NIR spectrometer and Micro-NIR 1700 spectrometer were compared with simulated fermentation process solutions. The Rc2, Rp2, RMSECV and RMSEP of the optimal FT-NIR and Micro-NIR 1700 models were 0.999, 0.999, 3.226 g/L, 1.388 g/L and 0.999, 0.999, 1.821 g/L, 0.967 g/L. Passing-Bablok regression method and paired t-test results showed there were no significant differences between the two instruments. Then the Micro-NIR 1700 was selected for the practical fermentation process, 135 samples from 10 batches were collected. Spectral pretreatment methods and variables selection methods (BiPLS, FiPLS, MWPLS and CARS-PLS) for PLS modeling were discussed. The Rc2, Rp2, RMSECV and RMSEP of the optimal GlcNAc content PLS model of the practical fermentation process were 0.994, 0.995, 2.792 g/L and 1.946 g/L. The results have a positive reference for application of the Micro-NIR spectrometer. To some extent, it could provide theoretical supports in guiding the microbial fermentation or the further assessment of bioprocess.

Mixomics analysis of Bacillus subtilis: effect of oxygen availability on riboflavin production

BackgroundRiboflavin, an intermediate of primary metabolism, is one kind of important food additive with high economic value. The microbial cell factory Bacillus subtilis has already been proven to possess significant importance for the food industry and have become one of the most widely used riboflavin-producing strains. In the practical fermentation processes, a sharp decrease in riboflavin production is encountered along with a decrease in the dissolved oxygen (DO) tension. Influence of this oxygen availability on riboflavin biosynthesis through carbon central metabolic pathways in B. subtilis is unknown so far. Therefore the unveiled effective metabolic pathways were still an unaccomplished task till present research work.ResultsIn this paper, the microscopic regulation mechanisms of B. subtilis grown under different dissolved oxygen tensions were studied by integrating 13C metabolic flux analysis, metabolomics and transcriptomics. It was revealed that the glucose metabolic flux through pentose phosphate (PP) pathway was lower as being confirmed by smaller pool sizes of metabolites in PP pathway and lower expression amount of ykgB at transcriptional level. The latter encodes 6-phosphogluconolactonase (6-PGL) under low DO tension. In response to low DO tension in broth, the glucose metabolic flux through Embden–Meyerhof–Parnas (EMP) pathway was higher and the gene, alsS, encoding for acetolactate synthase was significantly activated that may result due to lower ATP concentration and higher NADH/NAD+ ratio. Moreover, ResE, a membrane-anchored protein that is capable of oxygen regulated phosphorylase activity, and ResD, a regulatory protein that can be phosphorylated and dephosphorylated by ResE, were considered as DO tension sensor and transcriptional regulator respectively.ConclusionsThis study shows that integration of transcriptomics, 13C metabolic flux analysis and metabolomics analysis provides a comprehensive understanding of biosynthesized riboflavin’s regulatory mechanisms in B. subtilis grown under different dissolved oxygen tension conditions. The two-component system, ResD–ResE, was considered as the signal receiver of DO tension and gene regulator that led to differences between biomass and riboflavin production after triggering the shifts in gene expression, metabolic flux distributions and metabolite pool sizes.

PH and dose-dependent effects of quercetin on the fermentation capacity of Lactobacillus plantarum

This article reports a study of how quercetin affects the capacity of Lactobacillus plantarum RM71 to ferment different media, including a chemically defined medium (CDM) and media relevant for practical fermentation processes. It is shown for the first time that quercetin exerts pH- and dose-dependent effects on the fermentation performance of L. plantarum. At an initial pH of 5.5, quercetin promoted quicker growth upon inoculation at increased quercetin concentrations, while a detrimental dose-dependent lengthening of the lag phase was observed at an initial pH of 6.5. The time course of sugar consumption and lactic acid production data tracking in pH 5.5 CDM showed that quercetin promoted quicker sugar consumption as a result of earlier sugar uptake and lactic acid production than in the control. A model wine and a similar medium with modified sugar composition were fermented with L. plantarum RM71 on quercetin. Quercetin improved several key fermentation traits for the performance of L. plantarum in food production, including accelerated fermentation of various sugars, and accelerated malolactic fermentation and lactic acid production. Quercetin was not catabolized by L. plantarum in the fermentations, so the antioxidant properties of the flavonol were protected against degradation while the bacterium improved its growth performance.

Improvement of the fermentation performance of Lactobacillus plantarum by the flavanol catechin is uncoupled from its degradation

To determine the influence of the flavanol catechin on key metabolic traits for the fermentation performance of Lactobacillus plantarum strain RM71 in different media and to evaluate the ability of this strain to catabolize catechin. Growth monitoring and time course of sugar consumption data tracking in chemically defined medium (CDM), revealed that growth of Lact. plantarum strain RM71 upon catechin was characterized by a noticeable shorter lag period, outcome of earlier sugar consumption and lactic acid production courses. Catechin gave rise to higher cell densities compared to controls because of an increased extension of sugar utilization. Fermentation of media relevant for practical fermentation processes with Lact. plantarum strain RM71 showed that catechin sped up malic acid decarboxylation, which besides quicker and extended consumption of several sugars, resulted in faster and higher lactic acid production and growth. Spectrophotometric evaluation of catechin by HPLC-DAD and the lack of catechin concentration-dependent effects showed that the observed stimulations were uncoupled from catechin catabolism by Lact. plantarum. The flavanol catechin stimulated the growth of Lact. plantarum strain RM71 by promoting quicker sugar consumption, increasing the extension of sugar utilization and stimulating malic acid decarboxylation. These stimulations are uncoupled from catechin catabolism as Lact. plantarum did not catabolize it during fermentation. This study, for the first time, examined the influence of the flavanol catechin on the fermentation performance of a Lact. plantarum strain in several media under different fermentation conditions. The information could be relevant to control the production and obtain high-quality food products fermented by this micro-organism.

Microbial production of poly-D-3-hydroxybutyrate from CO2.

This short review covers the biotechnological aspects of the production of poly-D-3-hydroxybutyric acid, P(3HB), from H2, O2 and CO2 by autotrophic culture of the hydrogen-oxidizing bacterium, Ralstonia eutropha. Considering the efficiency of utilization of a gas mixture as substrate, a practical fermentation process using R. eutropha for the mass production of P(3HB) from CO2 should be designed on the basis of a recycled-gas, closed-circuit culture system. Also, maintaining the O2 concentration in a gas phase lower than 6.9% (v/v) is essential to prevent the gas mixture from exploding. Our study, using an explosion-proof fermentation bench plant and a two-stage culture system with a newly designed air-lift fermenter, demonstrated that very high P(3HB) yield and productivity could be obtained while the O2 concentration was maintained below 6.9%. However, a study on the continuous production of P(3HB) from CO2 by chemostat culture of R. eutropha revealed that the productivity and content of P(3HB) in the cells was considerably lower than by fed-batch culture. It is deduced that the use of the hydrogen-oxidizing bacterium, Alcaligenes latus, which accumulates P(3HB) even in the exponential growth phase, will be useful for the effective production of P(3HB) from CO2.

Safety assessment of genetically engineered yeast: elimination of mutagenicity of the yeast Saccharomyces cerevisiae by decreasing the activity of methylglyoxal synthase

Summary Methylglyoxal synthase catalyses the transformation of dihydroxyacetonephosphate to methylglyoxal, a toxic 2‐oxoaldehyde which is found to be present in the yeast Saccharomyces cerevisiae DKD‐5D‐H. Yeast cells in which genes for phosphoglucose isomerase, phosphofructokinase and triosephosphate isomerase had been extrachromosomally amplified by using a multi‐copy plasmid showed an increased ability to induce mutagenesis in standard tests when compared to wild type yeast. This response is mainly due to the increased amount of methylglyoxal in the engineered cells. To decrease the mutagenic activity and make it possible to use genetically engineered yeasts for practical fermentation processes, a mutant having a decreased level of methylglyoxal synthase activity was isolated. When transformed with genes for the glycolytic enzymes, the mutant cells showed extremely low levels of methylglyoxal content and mutagenic activity, both levels being comparable with those of non‐transformed DKD‐5D‐H cells.

Biosynthesis of D‐mannitol from D‐glucose by Aspergillus candidus

AbstractMannitol has long been known as a product of glucose metabolism by some strains of Aspergillus. Apparently no concerted effort, has been made to develop a practical fermentation process to make mannitol. Work at the Northern Laboratory has shown that nearly all strains of white Aspergillus produce significant amounts of mannitol; many strains of black Aspergillus also have this characteristic. Aspergillus candidus NRRL 305 is an exceptionally good mannitol producer. Studies on a fermentation process were conducted in 20‐1, stainless steel fermentors, without baffles. Czapek‐Dox medium, modified by addition of corn meal, yeast extract, and enzymatically hydrolyzed casein was the most satisfactory medium tested. Suitable increments of glucose were fed daily to the fermentors. The duration of the fermentation was from 10 to 16 days. The effects of agitation, aeration, temperature, and pH of the medium were studied. Under optimal conditions yields of mannitol approached 50% of the glucose consumed.