OPTIMIZATION OF BACTERIAL CELLULOSE PRODUCTION FROM SUGAR BEET MOLASSES BY GLUCONACETOBACTER XYLINUS NRRL B-759 IN STATIC CULTURE

We investigated the optimization of bacterial cellulose (BC) production from sugar beet molasses by Gluconacetobacter xylinus NRRL B-759 in static culture. The optimization studies were performed using the central composite design (CCD) of response surface methodology (RSM). The independent variables were the molasses concentration, inoculation ratio and culture volume. The dependent variable was BC production yield. From the optimization tests, based on the model developed by RSM-ANOVA, it was found that binary interactions between molasses concentration–culture volume and inoculation ratio–culture volume had the most significant influence on the responses. The optimum conditions were as follows: 78.932 g/L molasses concentration, 12.973% inoculation ratio, and 130.405 mL of culture volume. The obtained BC was characterized by Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM) and elemental analysis. The characterization results obtained in the study revealed that the produced BC exhibited typical FTIR spectrum, elemental composition, and nanofiber structure.

Bioreactor Suspension Culture: Differentiation and Production of Cardiomyocyte Spheroids From Human Induced Pluripotent Stem Cells

Human induced-pluripotent stem cells (hiPSCs) can be efficiently differentiated into cardiomyocytes (hiPSC-CMs) via the GiWi method, which uses small-molecule inhibitors of glycogen synthase kinase (GSK) and tankyrase to first activate and then suppress Wnt signaling. However, this method is typically conducted in 6-well culture plates with two-dimensional (2D) cell sheets, and consequently, cannot be easily scaled to produce the large numbers of hiPSC-CMs needed for clinical applications. Cell suspensions are more suitable than 2D systems for commercial biomanufacturing, and suspended hiPSCs form free-floating aggregates (i.e., spheroids) that can also be differentiated into hiPSC-CMs. Here, we introduce a protocol for differentiating suspensions of hiPSC spheroids into cardiomyocytes that is based on the GiWi method. After optimization based on cardiac troponin T staining, the purity of hiPSC-CMs differentiated via our novel protocol exceeded 98% with yields of about 1.5 million hiPSC-CMs/mL and less between-batch purity variability than hiPSC-CMs produced in 2D cultures; furthermore, the culture volume could be increased ∼10-fold to 30 mL with no need for re-optimization, which suggests that this method can serve as a framework for large-scale hiPSC-CM production.

Harvesting Marine Microalgae Nannochloropsis sp. using Dissolved Air Flotation (DAF) Technique

The production of high-value bioproducts from microalgae biomass has been widely investigated. However, their production is hindered by the expensive harvesting process. To date, flocculation followed by DAF process has been accepted as one of the affordable harvesting approaches. In this study, the use of DAF technique was attempted to harvest marine microalgae Nannochloropsis sp. Batch DAF harvesting was carried out using fabricated DAF unit equipped with several compartments including separation column, product collecting vessel and rotary skimmer. Tannin-based biopolymer flocculant, AFlok-BP1 at pH 5 with a concentration of 160 mg/L was used to facilitate the flocculation of particles. The effects of different saturator pressure at 1.8, 2, and 2.2 bar were then evaluated at a constant volume of 6 L microalgae culture. The effects of different microalgae culture volumes (6, 8 and 10 L) were also evaluated at a fixed saturator pressure of 2.2 bar. The highest pressure at 2.2 bar yielded the best result with the highest total solid of 3.19 ± 0.01% and a maximum yield of 1.70 ± 0.05 g/g (wet basis). The microalgae concentration was the lowest (0.027 g/L) when 6 L of culture volume was used. However, the values were significantly higher when the culture volume was increased to 8 and 10 L to approximately 0.035 and 0.050 g/L, respectively. As a conclusion, the study provided evidence for the feasibility of DAF technique in harvesting marine microalgae Nannochloropsis sp.

From Queen Anne to Queen Victoria. Readings in 18th and 19th century British literature and culture. Volume 7

The present, seventh, volume of the series "From Queen Anne to Queen Victoria" offers profitable re-visitations of old themes, as well as explorations of new themes and problems. The volume focuses on the literature, culture, and political and social history of Britain in a period when the structures of industrial modernity were being created, and examines Britain’s imprint on the global cultural heritage, including class, gender and race-based hierarchies that persist in varying degrees into the present.

Uncoupling Foam Fractionation and Foam Adsorption for Enhanced Biosurfactant Synthesis and Recovery

The production of biosurfactants is often hampered by excessive foaming in the bioreactor, impacting system scale-up and downstream processing. Foam fractionation was proposed to tackle this challenge by combining in situ product removal with a pre-purification step. In previous studies, foam fractionation was coupled to bioreactor operation, hence it was operated at suboptimal parameters. Here, we use an external fractionation column to decouple biosurfactant production from foam fractionation, enabling continuous surfactant separation, which is especially suited for system scale-up. As a subsequent product recovery step, continuous foam adsorption was integrated into the process. The configuration is evaluated for rhamnolipid (RL) or 3-(3-hydroxyalkanoyloxy)alkanoic acid (HAA, i.e., RL precursor) production by recombinant non-pathogenic Pseudomonas putida KT2440. Surfactant concentrations of 7.5 gRL/L and 2.0 gHAA/L were obtained in the fractionated foam. 4.7 g RLs and 2.8 g HAAs could be separated in the 2-stage recovery process within 36 h from a 2 L culture volume. With a culture volume scale-up to 9 L, 16 g RLs were adsorbed, and the space-time yield (STY) increased by 31% to 0.21 gRL/L·h. We demonstrate a well-performing process design for biosurfactant production and recovery as a contribution to a vital bioeconomy.

Culture Volume Influences the Dynamics of Adaptation under Long-Term Stationary Phase

Escherichia coli and many other bacterial species, which are incapable of sporulation, can nevertheless survive within resource exhausted media by entering a state termed long-term stationary phase (LTSP). We have previously shown that E. coli populations adapt genetically under LTSP in an extremely convergent manner. Here, we examine how the dynamics of LTSP genetic adaptation are influenced by varying a single parameter of the experiment—culture volume. We find that culture volume affects survival under LTSP, with viable counts decreasing as volumes increase. Across all volumes, mutations accumulate with time, and the majority of mutations accumulated demonstrate signals of being adaptive. However, positive selection appears to affect mutation accumulation more strongly at higher, compared with lower volumes. Finally, we find that several similar genes are likely involved in adaptation across volumes. However, the specific mutations within these genes that contribute to adaptation can vary in a consistent manner. Combined, our results demonstrate how varying a single parameter of an evolutionary experiment can substantially influence the dynamics of observed adaptation.