Integration of Aspergillus niger transcriptomic profile with metabolic model identifies potential targets to optimise citric acid production from lignocellulosic hydrolysate

Background Citric acid is typically produced industrially by Aspergillus niger-mediated fermentation of a sucrose-based feedstock, such as molasses. The fungus Aspergillus niger has the potential to utilise lignocellulosic biomass, such as bagasse, for industrial-scale citric acid production, but realising this potential requires strain optimisation. Systems biology can accelerate strain engineering by systematic target identification, facilitated by methods for the integration of omics data into a high-quality metabolic model. In this work, we perform transcriptomic analysis to determine the temporal expression changes during fermentation of bagasse hydrolysate and develop an evolutionary algorithm to integrate the transcriptomic data with the available metabolic model to identify potential targets for strain engineering. Results The novel integrated procedure matures our understanding of suboptimal citric acid production and reveals potential targets for strain engineering, including targets consistent with the literature such as the up-regulation of citrate export and pyruvate carboxylase as well as novel targets such as the down-regulation of inorganic diphosphatase. Conclusions In this study, we demonstrate the production of citric acid from lignocellulosic hydrolysate and show how transcriptomic data across multiple timepoints can be coupled with evolutionary and metabolic modelling to identify potential targets for further engineering to maximise productivity from a chosen feedstock. The in silico strategies employed in this study can be applied to other biotechnological goals, assisting efforts to harness the potential of microorganisms for bio-based production of valuable chemicals.

Review on production of citric acid by fermentation technology

Citric acid is the most important organic acid produced in tonnage and is extensively used in food and pharmaceutical industries. It is produced mainly by submerged fermentation using Aspergillus niger or Candida sp. from different sources of carbohydrates, such as molasses and starch-based media. In view of surges in demand and growing markets, there is always a need for the discovery and development of better production techniques and solutions to improve production yields and the efficiency of product recovery. To support the enormous scale of production, it is necessary and important for the production process to be environmentally friendly by utilizing readily available and inexpensive agro-industrial waste products, while maintaining high production yields. This review article for fermentation of citric acid and Microbial production of citric acid, Substrates and strategies of citric acid production for Surface fermentation, Submerged fermentation, Solid-state fermentation and also the effects of various Factors affecting of citric acid fermentation conditions are Carbon source, Nitrogen limitation, Phosphorus source, Lower Alcohols, pH of culture medium, Trace elements, Aeration and Other factors. citric acid recovery options and the numerous applications of citric acid, based on the literature review information of citric acid production by fermentation technology.

Enhanced Production of Citric Acid by Mutant PN12 of Aspergillus fumigatus Using Statistical Design

Distillery spent wash is an unwanted residual liquid waste generated during alcohol production. It is a potential source for production of different industrially important products. Distillery spent wash is dark colored and has many organic compounds as a waste. In this experiment, removal of color and organic compounds was carried out by anaerobic treatment. The treated spent wash was utilized for citric acid production with the help of microorganisms. The current study was performed with the treated spent wash which was applied for high level of citric acid production by a mutant strain of Aspergillus fumigatus PN12. The parent strain Aspergillus fumigatus PN12 was mutagenized by UV exposure to enhance citric acid production. After UV exposure investigation, mutant strain was selected for optimization and statistical method. The best citric acid production obtained was, 26.45 g/L at 30 ℃ with pH 6.0, 0.1 g/L of KH2PO4 and (NH4)2SO4 under OFAT. Under RSM optimization, maximum citric acid production was achieved as 30.89 g/L. Thus, the process optimization through the statistical approach resulted in a 1.16-fold enhancement in citric acid production as compared to that of the OFAT parametric conditions. Citric acid producing enzymes such as aconitase, NAD+-isocitrate dehydrogenase and NADP+ isocitrate dehydrogenase was studied. Maximum activity (U/mg) of aconitase (3.19±0.023), NAD+-isocitrate dehydrogenase (3.0±0.15) and NADP+ isocitrate dehydrogenase (2.91±0.17) was observed at 96 h. The present study can conclude that spent wash is potential source for citric acid production. Utilization of mutant strain of Aspergillus fumigatus PN12 is beneficiary for large scale industrial fermentation and citric acid production.

A Kinetic model for submerged citric acid production by Aspergillus versicolor using oil palm empty fruit bunch

The fermentation kinetics of citric acid by Aspergillus versicolor was studied in a submerged batch system. The logistic equation for growth, the Luedeking–Piret equation for citric acid production and modified Luedeking–Piret-like equation for glucose consumption was proposed for this study. The model appeared to provide a reasonable description for each parameter during the growth phase. The production of citric acid was growth-associated.

Biological production of citric acid in submerged culture of Aspergillus niger using cassava pulp wastes

Utilization of cassava pulp wastes for citric acid production was investigated using Aspergillus niger in a submerged culture. A series of experiments were designed on various fermentation parameters to establish the optimal conditions for citric acid production from cassava pulp. This study revealed that production parameters such as cassava pulp concentration, initial pH, incubation temperature, agitation, and nitrogen source and fermentation period had effect on the amount of citric acid produced from cassava pulp. Citric acid concentration increased as the concentration of cassava pulp increases up to 20% with maximum citric acid concentration of 14.9 ± 0.413 g/l after 120 hours of fermentation. pH 5.5 was the optimum with maximum citric acid concentration of 16.8 ± 0.23 g/l after 120 hours of fermentation. Incubation temperature at 300 C was the optimum, with citric acid concentration of 19.15 ± 0.43 g/l. Increased in agitation speed from 100 to 225 rpm gave the maximum citric acid concentration of 25.2 ± 0.32 g/l after 120 hours of fermentation. Soybean meal supplementation was the best maximum citric acid concentration of 28.2 ± 0.51 g/l. Evaluating the effect of different concentration of soybean meal shows that 0.3 % supplementation was the optimum with maximum concentration of 31.2 ±0.35g/l from cassava pulp after 120 hours of fermentation. The result suggested that citric acid can be accumulated using cassava pulp by Aspergillus niger in submerged culture during fermentation. Cassava pulp if well harnessed can be used for large scale citric acid production.

Improving citric acid production of an industrial Aspergillus niger CGMCC 10142: identification and overexpression of a high-affinity glucose transporter with different promoters

Background Glucose transporters play an important role in the fermentation of citric acid. In this study, a high-affinity glucose transporter (HGT1) was identified and overexpressed in the industrial strain A. niger CGMCC 10142. HGT1-overexpressing strains using the PglaA and Paox1 promoters were constructed to verify the glucose transporter functions. Result As hypothesized, the HGT1-overexpressing strains showed higher citric acid production and lower residual sugar contents. The best-performing strain A. niger 20-15 exhibited a reduction of the total sugar content and residual reducing sugars by 16.5 and 44.7%, while the final citric acid production was significantly increased to 174.1 g/L, representing a 7.3% increase compared to A. niger CGMCC 10142. Measurement of the mRNA expression levels of relevant genes at different time-points during the fermentation indicated that in addition to HGT1, citrate synthase and glucokinase were also expressed at higher levels in the overexpression strains. Conclusion The results indicate that HGT1 overexpression resolved the metabolic bottleneck caused by insufficient sugar transport and thereby improved the sugar utilization rate. This study demonstrates the usefulness of the high-affinity glucose transporter HGT1 for improving the citric acid fermentation process of Aspergillus niger CGMCC 10142.