Characterization of a symbiotic beverage based on water-soluble soybean extract fermented by Lactiplantibacillus plantarum ATCC 8014.

The health benefits of functional foods are associated with consumer interest and have supported the growth of the market for these types of foods, with emphasis on the development of new formulations based on plant extracts. Therefore, the present study aimed to characterize a symbiotic preparation based on water-soluble soy extract, supplemented with inulin and xylitol and fermented by Lactiplantibacillus plantarum ATCC 8014. Regarding nutritional issues, the symbiotic formulation can be considered a source of fiber (2 g/100 mL) and proteins (2.6 g/100 mL), and it also has a low-fat content and low caloric value. This formulation, in terms of microbiological aspects, remained adequate to legal standards after storage for 60 days under refrigeration and also presented an adequate quantity of the aforementioned probiotic strain, corresponding to 9.11 Log CFU.mL-1. These viable L. plantarum cells proved to be resistant to simulated human gastrointestinal tract conditions, reaching the intestine at high cell concentrations of 7.95 Log CFU.mL-1 after 60 days of refrigeration. Regarding sensory evaluation, the formulation showed good acceptance, presenting an average overall impression score of 6.98, 5.98, and 5.16, for control samples stored for 30 and 60 days under refrigeration, respectively. These results demonstrate that water-soluble soy extract is a suitable matrix for fermentation involving L. plantarum ATCC 8014, supporting and providing data on the first steps towards the development of a symbiotic functional food, targeting consumers who have restrictions regarding the consumption of products of animal origin, diabetics, and individuals under calorie restrictions.

Enzymatic potential of endophytic fungi: xylanase production by Colletotrichum boninense from sugarcane biomass.

Endophytic fungi constitute a major part of the still unexplored fungal diversity and have gained interest as new biological sources of natural active compounds, including enzymes. Endophytic fungi were isolated from soybean leaves and initially screened on agar plates for the production of CMCase (carboxymethylcellulase), xylanase, amylase and protease. The highest Enzymatic Indexes (IE) were verified for xylanase (2.14 and 1.31) with the fungi M6-A6P5F2 and M12-A5P3F1.2 and CMCase (1.92 and 1.62) with the fungi M13-A9P2F1 and M12-A5P3F1.2, respectively. The production of xylanase and CMCase by the selected fungi was evaluated in submerged cultivation using beechwood xylan and carboxymethylcellulose (CMC), as well as sugarcane straw and bagasse in different ratios as carbon sources. Both types of lignocellulosic biomass proved to be good inducers of enzymatic activity. The best xylanase producer among the isolates was identified as Colletotrichum boninense. With this fungus, the highest xylanase activity was obtained with a sugarcane straw-bagasse mixture in a 50:50 ratio (383.63 U mL-1), a result superior to that obtained with the use of beechwood xylan (296.65 U mL-1). Regardingthe kinetic behavior of the crude xylanase, there was found optimal pH of 5.0 and optimal temperatures of 50°C and 60°C. At 40°C and 50°C, xylanase retained 87% and 76% of its initial catalytic activity, respectively. These results bring new perspectives on bioprospecting endophytic fungi for the production of enzymes, mainly xylanase, as well as the exploitation of agro-industrial by-products, such as sugarcane straw and bagasse.

Valorization of apple pomace using bio-based technology for the production of xylitol and 2G ethanol.

Apple pomace was studied as a raw material for the production of xylitol and 2G ethanol, since this agroindustrial residue has a high concentration of carbohydrate macromolecules, but is still poorly studied for the production of fermentation bioproducts, such as polyols. The dry biomass was subjected to dilute-acid hydrolysis with H2SO4 to obtain the hemicellulosic hydrolysate, which was concentrated, detoxified and fermented. The hydrolyzate after characterization was submitted to submerged fermentations, which were carried out in Erlenmeyer flasks using, separately, the yeasts Candida guilliermondii and Kluyveromyces marxianus. High cellulose (32.62%) and hemicellulose (23.60%) contents were found in this biomass, and the chemical hydrolysis yielded appreciable quantities of fermentable sugars, especially xylose. Both yeasts were able to metabolize xylose, but Candida guilliermondii produced only xylitol (9.35 g L-1 in 96 h), while K. marxianus produced ethanol as the main product (10.47 g L-1 in 24 h) and xylitol as byproduct (9.10 g L-1 xylitol in 96 h). Maximum activities of xylose reductase and xylitol dehydrogenase were verified after 24 h of fermentation with C. guilliermondii (0.23 and 0.53 U/mgprot, respectively) and with K. marxianus (0.08 e 0.08 U/mgprot, respectively). Apple pomace has shown potential as a raw material for the fermentation process, and the development of a biotechnological platform for the integrated use of both the hemicellulosic and cellulosic fraction could add value to this residue and the apple production chain.

Xylitol bioproduction: state-of-the-art, industrial paradigm shift, and opportunities for integrated biorefineries.

Recent advances in biomass conversion technologies have shown a promising future toward fermentation during xylitol production. Xylitol is one of the top 12 renewable added-value chemicals that can be obtained from biomass according to US Department of Energy (USDOE). Currently, xylitol accounts for approximately US$823.6 million of annual sales in the market, and this amount is expected to reach US$1.37 billion by 2025. This high demand has been achieved owing to the chemical conversion of hemicellulosic hydrolysates from different lignocellulosic biomasses, which is a costly and non-ecofriendly process. Xylose-rich hemicellulosic hydrolysates are the major raw materials for xylitol production through either chemical or biotechnological routes. Economic production of a clean hemicellulosic hydrolysate is one of the major bottlenecks for xylitol production on the commercial scale. Advancements in biotechnology, such as the isolation of novel microorganisms, genetic manipulation of xylose metabolizing strains, and modifications in the fermentation process, can enhance the economic feasibility of xylitol production on the large scale. Furthermore, xylitol production in integrated biorefineries can be even more economic, given the readily available raw materials and the co-use of steam, electricity, and water, among others. Exploring new biotechnology techniques in integrated biorefineries would open new markets and opportunities for sustainable xylitol production to fulfill the market's growing demands for this sugar alcohol. This article is a review of the advancements reported in the whole biotechnological process for xylitol production, and involve pretreatment technologies, hemicellulosic hydrolysate preparation, xylose conversion into xylitol, and product recovery. Special attention is devoted to current metabolic engineering strategies to improve this bioprocess, as well as to the importance of xylitol production processes in biorefineries.

Metabolism of biodiesel-derived glycerol in probiotic Lactobacillus strains.

Three probiotic Lactobacillus strains, Lactobacillus acidophilus, Lactobacillus plantarum, and Lactobacillus delbrueckii, were tested for their ability to assimilate and metabolize glycerol. Biodiesel-derived glycerol was used as the main carbon and energy source in batch microaerobic growth. Here, we show that the tested strains were able to assimilate glycerol, consuming between 38 and 48 % in approximately 24 h. L. acidophilus and L. delbrueckii showed a similar growth, higher than L. plantarum. The highest biomass reached was 2.11 g L⁻¹ for L. acidophilus, with a cell mass yield (Y (X/S)) of 0.37 g g⁻¹. L. delbrueckii and L. plantarum reached a biomass of 2.06 and 1.36 g L⁻¹. All strains catabolize glycerol mainly through glycerol kinase (EC 2.7.1.30). For these lactobacillus species, kinetic parameters for glycerol kinase showed Michaelis-Menten constant (K(m)) ranging from 1.2 to 3.8 mM. The specific activities for glycerol kinase in these strains were in the range of 0.18 to 0.58 U mg protein⁻¹, with L. acidophilus ATCC 4356 showing the maximum specific activity after 24 h of cultivation. Glycerol dehydrogenase activity was also detected in all strains studied but only for the reduction of glyceraldehyde with NADPH (K(m) for DL-glyceraldehyde ranging from 12.8 to 32.3 mM). This enzyme shows a very low oxidative activity with glycerol and NADP+ and, most likely, under physiological conditions, the oxidative reaction does not occur, supporting the assumption that the main metabolic flux concerning glycerol metabolism is through the glycerol kinase pathway.

Obtaining partial purified xylose reductase from Candida guilliermondii.

The enzymatic bioconversion of xylose into xylitol by xylose reductase (XR) is an alternative for chemical and microbiological processes. The partial purified XR was obtained by using the following three procedures: an agarose column, a membrane reactor or an Amicon Ultra-15 50K Centrifugal Filter device at yields of 40%, 7% and 67%, respectively.