A suitable nanofiber sheet was formulated and developed based on its efficacy in the immobilization of recombinant Escherichia coli (E. coli) to enhance xylitol production. The effects of different types of nanofibers and solvents on cell immobilization and xylitol production were studied. The most applicable nanofiber membrane was selected via preliminary screening of four types of nanofiber membrane, followed by the selection of six different solvents. Polyvinylidene fluoride (PVDF) nanofiber sheet synthesized using dimethylformamide (DMF) solvent was found to be the most suitable carrier for immobilization and xylitol production. The thin, beaded PVDF (DMF) nanofibers were more favourable for microbial adhesion, with the number of immobilized cells as high as 96 × 106 ± 3.0 cfu/ml. The attraction force between positively charged PVDF nanofibers and the negatively charged E. coli indicates that the electrostatic interaction plays a significant role in cell adsorption. The use of DMF has also produced PVDF nanofibers biocatalyst capable of synthesizing the highest xylitol concentration (2.168 g/l) and productivity (0.090 g/l/h) and 55-69% reduction in cell lysis compared with DMSO solvent and free cells. This finding suggests that recombinant E. coli immobilized on nanofibers shows great potential as a whole-cell biocatalyst for xylitol production.
E. coli has been engineered to produce xylitol, but the production faces bottlenecks in terms of production yield and cell viability. In this study, recombinant E. coli (rE. coli) was immobilized on untreated and treated multiwalled carbon nanotubes (MWCNTs) for xylitol production. The immobilized rE. coli on untreated MWCNTs gave the highest xylitol production (5.47 g L-1) and a productivity of 0.22 g L-1 h-1. The doubling time for the immobilized cells increased up to 20.40 h and was higher than that of free cells (3.67 h). Cell lysis of the immobilized cells was reduced by up to 73 %, and plasmid stability improved by up to 17 % compared to those of free cells. Xylitol production using the optimum parameters (pH 7.4, 0.005 mM and 29 °C) achieved a xylitol production and productivity of 6.33 g L-1 and 0.26 g L-1 h-1, respectively. A seven-cycle repeated batch fermentation was carried out for up to 168 h, which showed maximum xylitol production of 7.36 g L-1 during the third cycle. Hence, this new adsorption immobilization system using MWCNTs is an alternative to improve the production of xylitol.
Kenaf (Hibiscus cannabinus L.), a potential fibre crop with a desirably high growth rate, could serve as a sustainable feedstock in the production of xylitol. In this work, the extraction of soluble products of kenaf through dilute nitric-acid hydrolysis was elucidated with respect to three parameters, namely temperature, residence time, and acid concentration. The study will assist in evaluating the performance in terms of xylose recovery. The result point out that the maximum xylose yield of 30.7 g per 100 g of dry kenaf was attained from 2% (v/v) HNO3 at 130 °C for 60 min. The detoxified hydrolysate was incorporated as the primary carbon source for subsequent fermentation by recombinant Escherichia coli and the performance of strain on five different semi-synthetic media on xylitol production were evaluated herein. Among these media, batch cultivation in a basal salt medium (BSM) afforded the highest xylitol yield of 0.35 g/g based on xylose consumption, which corresponded to 92.8% substrate utilization after 38 h. Subsequently, fermentation by E. coli in the xylose-based kenaf hydrolysate supplemented with BSM resulting in 6.8 g/L xylitol which corresponding to xylitol yield of 0.38 g/g. These findings suggested that the use of kenaf as the fermentation feedstock could be advantageous for the development of sustainable xylitol production.