Fermentative processes for the upcycling of xylose to xylitol by immobilized cells of Pichia fermentans WC1507.

Xylitol is a pentose-polyol widely applied in the food and pharmaceutical industry. It can be produced from lignocellulosic biomass, valorizing second-generation feedstocks. Biotechnological production of xylitol requires scalable solutions suitable for industrial scale processes. Immobilized-cells systems offer numerous advantages. Although fungal pellet carriers have gained attention, their application in xylitol production remains unexplored. In this study, the yeast strain P. fermentans WC 1507 was employed for xylitol production. The optimal conditions were observed with free-cell cultures at pH above 3.5, low oxygenation, and medium containing (NH4)2SO4 and yeast extract as nitrogen sources (xylitol titer 79.4 g/L, YP/S 66.3%, and volumetric productivity 1.3 g/L/h). Yeast cells were immobilized using inactive Aspergillus oryzae pellet mycelial carrier (MC) and alginate beads (AB) and were tested in flasks over three consecutive production runs. Additionally, the effect of a 0.2% w/v alginate layer, coating the outer surface of the carriers (cMC and cAB, respectively), was examined. While YP/S values observed with both immobilized and free cells were similar, the immobilized cells exhibited lower final xylitol titer and volumetric productivity, likely due to mass transfer limitations. AB and cAB outperformed MC and cMC. The uncoated AB carriers were tested in a laboratory-scale airlift bioreactor, which demonstrated a progressive increase in xylitol production in a repeated batch process: in the third run, a xylitol titer of 63.0 g/L, YP/S of 61.5%, and volumetric productivity of 0.52 g/L/h were achieved. This study confirmed P. fermentans WC 1507 as a promising strain for xylitol production in both free- and entrapped-cells systems. Considering the performance of the wild strain, a metabolic engineering intervention aiming at further improving the efficiency of xylitol production could be justified. MC and AB proved to be viable supports for cell immobilization, but additional process development is necessary to identify the optimal bioreactor configuration and fermentation conditions.

Insights into differences between spore-assisted and pellet-assisted microalgae harvesting using a highly efficient fungus: Efficiency, high-value substances, and mechanisms.

To achieve efficient and low-cost microalgae harvesting, investigations were conducted on the harvesting efficiency and potential mechanisms of Chlorella sp. HQ by filamentous fungi using two strategies, fungal spore-assisted harvesting (FSH) and fungal pellet-assisted harvesting (FPH). Five of the 19 fungal species isolated from domestic sewage could form pellets, and Aspergillus niger HW8-1 comprised the highest harvesting efficiencies of 99.17 % and 88.70 % for FPH and FSH, respectively. FSH had 2-3 times more lipids and polysaccharides in fungus-alga pellets and caused richer saturated and monounsaturated fatty acids compared with FPH. Moreover, by optimizing the microalgae preculture time, glucose concentration, and microalgae initial density, the contents of high-value substances, such as lipids, polysaccharides, and proteins of fungus-alga pellets after FPH were improved from 5.96 %, 5.67 %, and 7.27 % to 20.18 %, 24.34 % and 10.48 %, respectively. Furthermore, fungal pellets secreted more extracellular polymeric substances (EPS) during FPH than those by FSH, which could chemisorb algal cells by lowering the surface potential of fungal pellets. FPH caused algal cells to cover the outside, which increased the light exposure area of algae, thereby increasing the photosynthesis rate, whereas FSH mainly captured microalgae physically through mycelium entanglement.

Biochar fungal pellet based biological immobilization reactor efficiently removed nitrate and cadmium.

To efficiently and simultaneously remove nitrate (NO3--N) and Cd(II) from aqueous solution, a novel type of biochar fungal pellet (BFP) immobilized denitrification bacteria (Cupriavidus sp. H29) composite was used in a bioreactor. The removal performance of the bioreactor R1 for the initial concentration of 27.7 mg L-1 nitrate and 10.0 mg L-1 Cd(II) reached 98.1 and 93.9% respectively, and the inoculation of strain H29 in bioreactor R1 significantly enhanced the removal efficiency of contaminants. The 3D-EEM spectra analysis showed that the activity of microorganisms in the bioreactor was higher at a lower concentration of Cd(II). FTIR indicated the effect of functional groups in BFP in bioadsorption of Cd(II). In addition, high-throughput analysis of species composition of the microbial community in the bioreactors at different levels demonstrated that strain H29 played a significant part in the bioreactor. This research provided a perspective for simultaneous restoration of nitrate and heavy metals in wastewater, and also enriched the application of fungal pellet (FP) in reactors.

Self-immobilized biochar fungal pellet combined with bacterial strain H29 enhanced the removal performance of cadmium and nitrate.

A newly isolated strain Phoma sp. ZJ6, which could form fungal pellet (FP) by self-immobilization, was identified. A novel longan seed biochar embedded in FP (BFP) combined with strain H29 (BFP-H29) effectively improved the Cd(II) removal efficiency and simultaneously removed nitrate. The adsorption process of BFP was well fitted with the pseudo-second-order kinetics model and Langmuir isotherm model, which demonstrated that the adsorption process was favorable and mainly dominated by chemisorption. Compared with single FP, biochar, and strain H29, BFP-H29 significantly enhanced the Cd(II) removal and the removal ratio reached 90.47%. Meanwhile, the simultaneous removal efficiency of the BFP-H29 for nitrate could reach 93.80%. Characterization analysis demonstrated that the primary removal mechanisms of BFP-H29 were precipitation and surface complexation. BFP-H29 had excellent performance in simultaneous removal of Cd(II) and nitrate, indicating its potential as a promising composite in the removal of cadmium and nitrate in wastewater.

Fungal pellets immobilized bacterial bioreactor for efficient nitrate removal at low C/N wastewater.

In this study, fungal pellets immobilized denitrifying Pseudomonas stutzeri sp. GF3 was cultivated to establish a bioreactor. The denitrification effect of fixed bacteria with fungal pellets was tested by response surface methodology (RSM). Analysis of the bioreactor showed that the denitrification efficiency reached 100% under the optimal conditions and the denitrification efficiency of the actual wastewater treatment in the stable phase reached 95.91%. Moreover, the organic matter and functional groups in the bioreactor under different C/N conditions were analyzed by fluorescence excitation-emission matrix (EEM) spectra and Fourier transform infrared spectroscopy (FTIR), which revealed that metabolic activities of denitrifying bacteria were enhanced with the increase of C/N. The morphology and structure of bacteria immobilized by fungal pellets explored by scanning electron microscope (SEM) showed the filamentous porous fungal pellets loaded with bacteria. Community structure analysis by high-throughput sequencing demonstrated that strain GF3 might was the dominant strain in bioreactor.

Selenite removal from wastewater using fungal pelleted airlift bioreactor.

This study investigated the removal of selenite from wastewater using the fungus Asergillus niger KP isolated from a laboratory scale inverse fluidized bed bioreactor. The effect of different carbon sources and initial selenite concentration on fungal growth, pellet formation and selenite removal was first examined in a batch system. The fungal strain showed a maximum selenite removal efficiency of 86% in the batch system. Analysis of the fungal pellets by field-emission scanning electron microscopy, field-emission transmission electron microscopy and energy-dispersive X-ray spectroscopy revealed the formation of spherical-shaped elemental selenium nanoparticles of size 65-100 nm. An increase in the initial selenite concentration in the media resulted in compact pellets with smooth hyphae structure, whereas the fungal pellets contained hair like hyphae structure when grown in the absence of selenite. Besides, a high initial selenite concentration reduced biomass growth and selenite removal from solution. Using an airlift reactor with fungal pellets, operated under continuous mode, a maximum selenite removal of 94.3% was achieved at 10 mg L-1 of influent selenite concentration and 72 h HRT (hydraulic retention time). Overall, this study demonstrated very good potential of the fungal-pelleted airlift bioreactor system for removal of selenite from wastewater. Graphical abstract.

The filamentous fungal pellet-relationship between morphology and productivity.

Filamentous fungi are used for the production of a multitude of highly relevant biotechnological products like citric acid and penicillin. In submerged culture, fungi can either grow in dispersed form or as spherical pellets consisting of aggregated hyphal structures. Pellet morphology, process control and productivity are highly interlinked. On the one hand, process control in a bioreactor usually demands for compact and small pellets due to rheological issues. On the other hand, optimal productivity might be associated with less dense and larger morphology. Over the years, several publications have dealt with aforementioned relations within the confines of specific organisms and products. However, contributions which evaluate such interlinkages across several fungal species are scarce. For this purpose, we are looking into methods to manipulate fungal pellet morphology in relation to individual species and products. This review attempts to address (i) how variability of pellet morphology can be assessed and (ii) how morphology is linked to productivity. Firstly, the mechanism of pellet formation is outlined. Subsequently, the description and analysis of morphological variations are discussed to finally establish interlinkages between productivity, performance and morphology across different fungal species.

Morphological Characteristic Regulation of Ligninolytic Enzyme Produced by Trametes polyzona.

A newly isolated white rot fungal strain KU-RNW027 was identified as Trametes polyzona, based on an analysis of its morphological characteristics and phylogenetic data. Aeration and fungal morphology were important factors which drove strain KU-RNW027 to secrete two different ligninolytic enzymes as manganese peroxidase (MnP) and laccase. Highest activities of MnP and laccase were obtained in a continuous shaking culture at 8 and 47 times higher, respectively, than under static conditions. Strain KU-RNW027 existed as pellets and free form mycelial clumps in submerged cultivation with the pellet form producing more enzymes. Fungal biomass increased with increasing amounts of pellet inoculum while pellet diameter decreased. Strain KU-RNW027 formed terminal chlamydospore-like structures in cultures inoculated with 0.05 g/L as optimal pellet inoculum which resulted in highest enzyme production. Enzyme production efficiency of T. polyzona KU-RNW027 depended on fungal pellet morphology as size, porosity, and formation of chlamydospore-like structures.

Morphological Characteristic Regulation of Ligninolytic Enzyme Produced by Trametes polyzona

A newly isolated white rot fungal strain KU-RNW027 was identified as Trametes polyzona, based on an analysis of its morphological characteristics and phylogenetic data. Aeration and fungal morphology were important factors which drove strain KU-RNW027 to secrete two different ligninolytic enzymes as manganese peroxidase (MnP) and laccase. Highest activities of MnP and laccase were obtained in a continuous shaking culture at 8 and 47 times higher, respectively, than under static conditions. Strain KU-RNW027 existed as pellets and free form mycelial clumps in submerged cultivation with the pellet form producing more enzymes. Fungal biomass increased with increasing amounts of pellet inoculum while pellet diameter decreased. Strain KU-RNW027 formed terminal chlamydospore-like structures in cultures inoculated with 0.05 g/L as optimal pellet inoculum which resulted in highest enzyme production. Enzyme production efficiency of T. polyzona KU-RNW027 depended on fungal pellet morphology as size, porosity, and formation of chlamydospore-like structures.