Degradation of environment is a challenge to crop production around the world. Biological control of various plant diseases using antagonistic bacteria is an encouraging alternative to traditionally used chemical control strategies. Chitosan as a well-known natural flocculation agent also exhibits antimicrobial activity. The goal of this study was to investigate a dual nature of chitosan in flocculation of Bacillus sp. BioSol021 cultivation broth intended for biocontrol applications. Experiments were performed based on L₁₈ standard Taguchi orthogonal array design with five input parameters (chitosan type and dosage, pH value, rapid and slow mixing rates). In this study, the grey relational analysis was used to perform multi-objective optimization of the chosen responses, i.e., flocculation efficiency and four inhibition zone diameters against the selected phytopathogens. The results have indicated a great potential of a highly efficient method for removal of the Bacillus bacteria from the cultivation broth using chitosan. The good flocculation efficiency and high precipitate antimicrobial activity against the selected phytopathogens were achieved. It has been shown that multiple flocculation performance parameters were improved, resulting in slightly improved response values.

Food industry effluents represent one of the major concerns when it comes to environmental impact; hence, their valorization through different chemical and biological routes has been suggested as a possible solution. The vast amount of organic and inorganic nutrients present in food industry effluents makes them suitable substrates for microbial growth. This study suggests two valorization routes for whey as dairy industry effluent and flotation wastewater from the wine industry through microbial conversion to biocontrol agents as value-added products. Cultivations of the biocontrol strain Bacillus sp. BioSol021 were performed in a 16 L bioreactor to monitor the bioprocess course and investigate bioprocess kinetics in terms of microbial growth, sugar substrate consumption and surfactin synthesis, as an antimicrobial lipopeptide. The produced biocontrol agents showed high levels of biocontrol activity against mycotoxigenic strains of Aspergillus flavus, followed by a significant reduction of sugar load of the investigated effluents by the producing microorganisms. With proven high potential of whey and winery flotation wastewater to be used as substrates for microbial growth, this study provides grounds for further optimization of the suggested valorization routes, mostly in terms of bioprocess conditions to achieve maximal techno-economical feasibility, energy saving and maximal reduction of effluents’ organic and inorganic burden.

Production of highly efficient biomass-based microbial biopesticides significantly depends on downstream processing in terms of obtaining as high concentration of viable cells as possible. Microfiltration is one of the recommended operations for microbial biomass separation, but its main limitation is permeate flux decrease due to the membrane fouling. The effect of air sparging as a hydrodynamic technique for improvement of permeate flux during microfiltration of Bacillus velezensis cultivation broth was investigated. Modeling of the microfiltration was performed using the response surface methodology, while desirability function approach and genetic algorithm were applied for optimization, i.e., maximization of permeate flux and minimization of specific energy consumption. The results have revealed antagonistic relationship between the investigated dependent variables. The optimized values of superficial feed velocity and transmembrane pressure were close to the mean values of the investigated value ranges (0.68 bar and 0.96 m/s, respectively), while the optimized value of superficial air velocity had a more narrow distribution around 0.25 m/s. The results of this study have revealed a significant improvement of microfiltration performance by applying air sparging, thus this flux improvement method should be further investigated in downstream processing of different bacterial cultivation broths.

The downstream processing of efficient biomass-based microbial biopesticides is heavily reliant on obtaining the largest concentration of viable cells in the most cost-effective manner. The goal of this research was to assess the ability of chitosan flocculation to recover bacterial Bacillus sp. BioSol021 biomass from the broth after biological treatment of wastewaters from the dairy and wine industries. Second-order factorial design models were used to estimate the effect of chitosan concentration and mixing speed on flocculation efficiency, settling velocity, and antimicrobial activity against Aspergillus flavus, i.e., inhibition zone diameter. Response surface methodology was followed by multi-objective optimization by applying the desirability function (DF) and genetic algorithm (GA). The optimum values for flocculation efficiency, settling velocity, and inhibition zone diameter for cheese whey effluent were 88%, 0.10 mm/s, and 51.00 mm, respectively. In the case of winery flotation effluent, the optimum values were flocculation efficiency 95% and settling velocity 0.05 mm/s, while the inhibition zone diameter was 48.00 mm. These results indicate that utilizing chitosan as a flocculation agent not only fits the criteria for effective downstream processing, but also has a synergistic effect on Bacillus sp. antibacterial activity.

The use of membrane filtration as a downstream process for microbial biomass harvesting is hampered due to the low permeate flux values achieved during the microfiltration of fermentation broths. Several hydrodynamic methods for increasing permeate flux by creating turbulent flow patterns inside the membrane module are used to overcome this problem. The main goal of this study was to investigate the combined use of a Kenics static mixer and gas sparging during cross-flow microfiltration of Bacillus velezensis IP22 cultivation broth. Optimization of the microfiltration process was performed by using the response surface methodology. It was found that the combined use of a static mixer and gas sparging leads to a considerable increase in the permeate flux, up to the optimum steady-state permeate flux value of 183.42 L·m⁻²·h⁻¹ and specific energy consumption of 0.844 kW·h·m⁻³. The optimum steady-state permeate flux is almost four times higher, whilst, at the same time, the specific energy consumption is almost three times lower compared to the optimum results achieved using gas sparging alone. The combination of Kenics static mixer and gas sparging during cross-flow microfiltration is a promising technique for the enhancement of steady-state permeate flux with simultaneously decreasing specific energy consumption.

Eco-friendly pest management solutions are acknowledged as a crucial element in shaping the future of agriculture through sustainable practices. Achieving the maximum viable cell concentration while being cost-effective is the main goal of the downstream processing for efficient biomass-based microbial biopesticide production. The purpose of this study was to determine the effectiveness of chitosan flocculation in recovering bacterial Bacillus sp. BioSol021 biomass from broth cultivated using fruit juice industrial effluent as a medium, with the hypothesis of the synergistic effect of microbial and biopolymer components in phytopathogen suppression. Second-order polynomial models were used to calculate the influence of chitosan concentration and mixing speed on flocculation efficiency, settling velocity, and antibacterial activity against Aspergillus flavus (i.e., the inhibition zone diameter). The response surface approach, followed by desirability function optimization and the genetic algorithm were applied. The optimal values achieved in this study were 97.18%, 0.0369 mm/s, and 74.00 mm for flocculation efficiency, settling velocity, and inhibition zone diameter, respectively. The obtained results suggest that chitosan can be used as a flocculation agent for effective downstream processing, but also has a positive effect on the final product antimicrobial activity.