The enzymatic esterification of 5-hydroxymethylfurfural (HMF) with long-chain fatty acids offers a sustainable route for producing biolubricants and other high-value chemicals. This work evaluates the synthesis of 5-hidroxymethylfurfural stearate catalyzed by immobilized lipases in both batch and continuous packed-bed bioreactors, combining molecular dynamics (MD) simulations with experimental validation to identify suitable green solvents. Four solvents were tested: 2-methyl-3-buten-2-ol (2-MB), tert-butanol (TB), 2-methyltetrahydrofuran (2-MeTHF), and cyclopentyl methyl ether (CPME). MD simulations revealed that CPME increased hydrophobic surface exposure and flexibility near the catalytic site, favoring substrate accessibility. In preliminary experimental tests, CPME provided the highest conversion (50

Sustainability represents a significant global challenge, requiring a balance between environmental impact and the use of natural resources. White biotechnology, which uses microorganisms and enzymes for environmentally friendly products and processes, offers promising solutions to support a growing population. Within this context, the yeast Yarrowia lipolytica stands out, so we investigated the generation of biomass from two wild strains (ATCC 9773 and NRRL Y-50997) using different carbon sources. Additionally, protein content and amino acid profiles were assessed via standardized analytical methods to evaluate their potential as nutritional yeasts. Both strains demonstrated potential as nutritional yeasts, with biomass productivities of up to 35.5 g/L and 42 g/L, respectively. The protein content was high, with 58.8% for ATCC 9773 and 58.2% for NRRL Y-50997. Furthermore, the strains presented essential amino acid contents of 62.6% and 41.5%, with lysine being the most abundant amino acid. These findings underscore the versatility and productivity of Y. lipolytica, highlighting its potential for sustainable biotechnological applications such as single-cell protein production.

This study investigates the transesterification of isoflavones puerarin and beta-D-fructofuranosyl(2 -> 6)-puerarin to enhance their lipophilicity for specific pharmacological applications. The process achieved high yields (80-100%) in the isoflavones conversion when commercial lipases from Candida antarctica (CAL B) and Thermomyces lanuginosus ( TL IM) were used. The acylation patterns between CAL B and TL IM varied depending on the substrate; both enzymes yielded principally monoacylated products from puerarin and beta-D-fructofuranosyl-(2 -> 6)-puerarin. However, TL IM also produced di- and triacylated derivatives of beta-D-fructofuranosyl-(2 -> 6)-puerarin in substantial amounts. Structural characterization by NMR identified puerarin-6''- O-laurate and beta-D-fructofuranosyl-(2 -> 6)-puerarin-6'''-O-laurate as the primary products in CAL B and TL IM reactions. These acyl derivatives presented improved lipophilicity (Log P from -1.0 - 0.16 to 3.26 - 3.58) and antioxidant activity (67.2-97.3 to 102.5-131.4 mu M TE/mmol). Therefore, these compounds have potential pharmacological applications.

The extremotolerant red yeast Rhodotorula mucilaginosa displays resilience to diverse environmental stressors, including cold, osmolarity, salinity, and oligotrophic conditions. Particularly, this yeast exhibits a remarkable ability to accumulate lipids and carotenoids in response to stress conditions. However, research into lipid biosynthesis has been hampered by limited genetic tools and a scarcity of studies on adaptive responses to nutrient stressors stimulating lipogenesis. This study investigated the impact of nitrogen stress on the adaptive response in Antarctic yeast R. mucilaginosa M94C9. Varied nitrogen availability reveals a nitrogen-dependent modulation of biomass and lipid droplet production, accompanied by significant ultrastructural changes to withstand nitrogen starvation. In silico analysis identifies open reading frames of genes encoding key lipogenesis enzymes, including acetyl-CoA carboxylase (Acc1), fatty acid synthases 1 and 2 (Fas1/Fas2), and acyl-CoA diacylglycerol O-acyltransferase 1 (Dga1). Further investigation into the expression profiles of RmACC1, RmFAS1, RmFAS2, and RmDGA1 genes under nitrogen stress revealed that the prolonged up-regulation of the RmDGA1 gene is a molecular indicator of lipogenesis. Subsequent fatty acid profiling unveiled an accumulation of oleic and palmitic acids under nitrogen limitation during the stationary phase. This investigation enhances our understanding of nitrogen stress adaptation and lipid biosynthesis, offering valuable insights into R. mucilaginosa M94C9 for potential industrial applications in the future.

Background: Naringin is one of the main flavonoids in citrus fruits and byproducts. This flavanone has been shown to be a good antioxidant nutraceutical component, and it also has potential as a gut microbiome modulator, although its applications in final formulations represent a challenge due to its low solubility, both in water and in organic solvents. This work addresses this problem by functionalizing naringin through enzymatic acylation. Results: The enzymatic acylation catalyzed by the lipase Novozym (R) 435 and using acyl donors of different chain lengths, acetate (C2), propionate (C3), and laurate (C12), yielded in conversions of 95% at 24 h and 100% at 48 h, generating a monoacylated product. Both the aqueous and solvent solubility of acylated naringin products were improved while maintaining or even increasing their antioxidant activity. Conclusions: This acylation process significantly enhanced both the water and solvent solubility of the acylated naringin products while preserving or even enhancing their antioxidant activity. In addition to the gut -modulating properties of flavonoids, acylating them with short- and medium -chain fatty acids could enhance their potential applications in the emerging field of research dedicated to understanding and modulating gut health. How to cite: Gutierrez-Navarro E, Padilla -de la Rosa JD, Macias A, et al. Enzymatically acylated naringin with gut modulation potential. Electron J Biotechnol 2024; 68. https://doi.org/10.1016/j.ejbt.2023.12.003. (c) 2023 The Authors. Pontificia Universidad Catolica de Valparaiso. Production and hosting by Elsevier B. V. This is an open access article under the CC BY -NC -ND license (http://creativecommons.org/licenses/bync-nd/4.0/).

Metabolic syndrome (MetS) comprises a cluster of metabolic risk factors, which include obesity, hypertriglyceridemia, high blood pressure, and insulin resistance. The purpose of this study was to evaluate the effects of laurate-bioconjugated fructans on pro- and anti-inflammatory cytokines in Wistar rats with MetS induced by a high-fat diet. Laurate-bioconjugated fructans were synthesized with agave fructans, immobilized lipase B, and vinyl laureate as the acylant. Groups were fed a standard diet (NORMAL), a high-fat diet (HFD), or a high-fat diet plus laurate-bioconjugated fructans (FL PREV) for 9 weeks. A fourth group received a high-fat diet for 6 weeks, followed by simultaneous exposure to a high-fat diet and laurate-bioconjugated fructans for 3 additional weeks (FL REV). The dose of laurate-bioconjugated fructans was 130 mg/kg. Laurate-bioconjugated fructans reduced food and energy intake, body weight, body mass index, abdominal circumference, adipose tissue, adipocyte area, serum triglycerides, insulin, insulin resistance, and C-reactive protein but they increased IL-10 protein serum levels and mRNA expression. The impact of laurate-bioconjugated fructans on zoometric and metabolic parameters supports their potential as therapeutic agents to improve obesity, obesity comorbidities, insulin resistance, type 2 diabetes mellitus, and MetS.

The International Energy Agency Bioenergy Task 42 defined biorefinery as “the sustainable processing of biomass into a spectrum of bio-based products (food, feed, chemicals, materials) and energy (fuels, power, heat)”. This includes biochemical, thermochemical, chemical, and mechanical processes. Yeast and fungi are ideal agents for biochemical biorefineries because of their metabolic versatility. This chapter will focus on yeast-biorefineries which can use inexpensive agro-industrial waste to obtain two main products: bio-oils (feed and food), and the enzyme lipase, which is a high-value product that can modify the bio-oil to get more products. The so-called “unconventional yeasts” stand out for having the ability to metabolize a variety of carbon sources. Yarrowia lipolytica, Cutaneotrichosporon oleaginosus and Rhodotorula toruloides have been described as the most efficient in terms of yield of bio-oil. For instance, Y. lipolytica can consume glycerol to produce bio-oil, lipases, organic acids, polyols and single-cell protein. Its genome is known and can be modified to overproduce the target compound or to engineer metabolic routes for other products of interest such as polyols. Therefore, this yeast has been proposed as a model for yeast-biorefineries. Besides, enzymes are one of the products with more added value that can be produced in a biorefinery. Yeasts are also very good enzyme producers or even hosts for heterologous expression of enzymes. Moreover, solid fermentation is a configuration particularly suitable to use agro-wastes in a biorefinery. Indeed, fermented solids of agro-wastes may be directly employed as biocatalysts allowing the production of more cost-competitive bio-oil fuels and biobased chemicals. This strategy may use inexpensive agro-industrial wastes as solid support/substrates for microorganism growth and lipase production, avoiding expensive steps of enzyme purification and immobilization. Recombinant lipase from Thermomyces lanuginosus produced by solid-state fermentation using agro-industrial wastes and its application to obtain biodiesel is described as example.

Background: GDSL-like esterase/lipase proteins (GELPs) are enzymes that possess unique characteristics, they contain four invariable catalytic residues. Advances in the study of these proteins are interesting. The cloning and functional expression of a papaya esterase have not been reported. Therefore, in this work we evaluated the heterologous production of Carica papaya esterase CpEST in the yeast Komogataella phaffii (Pichia pastoris). Results: The cloning and expression of the protein was performed under the PAOX1 promoter, and productions of up to 43 AU/mL were achieved using residual glycerol from biodiesel in the batch phase and methanol for the induction phase. Enzyme activity assays determined that CpEST has a high preference for short-chain substrates (p-NP C4 and p-NP C8), and optimal activity conditions were observed at 30 degrees C and pH 10. The enzyme showed the highest stability to acetone, ethanol and tert-butanol solvents, retaining approximately 55% of its initial enzymatic activity after 1 h of exposure. Conclusions: Cloning and functional expression of papaya CpEST esterase was achieved. During fermentation, the yeasts used as a carbon source residual glycerol from biodiesel production. Based on the results obtained from the characterization of the esterase, it was found that it has a high potential for use in the bioenergy and detergent industry. How to cite: Reyes-Reyes AL, Valero F, Sandoval G. Cloning, protein expression and biochemical characterization of Carica papaya esterase. Electron J Biotechnol 2022;61. https://doi.org/10.1016/j.ejbt. 2022.11.004. (c) 2022 Pontificia Universidad Catolica de Valparaiso. Production and hosting by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Metabolic syndrome is a complex disorder that combines abdominal obesity, dyslipidemia, hypertension, and insulin resistance. Metabolic syndrome affects 25% of the world's population. Agave fructans have shown positive effects on alterations related to metabolic syndrome, so some investigations have focused on their bioconjugation with fatty acids to increase their biological activity. The objective of this work was to evaluate the effect of agave fructan bioconjugates in a rat model with metabolic syndrome. Agave fructans enzymatically bioconjugated (acylated via food-grade lipase catalysis) with propionate or laurate were administered orally for 8 weeks in rats fed a hypercaloric diet. Animals without treatment were used as the control group, as well as animals fed with a standard diet. The data indicate that the group of animals treated with laurate bioconjugates showed a significant decrease in glucose levels, systolic pressure, weight gain, and visceral adipose tissue, as well as a positive effect of pancreatic lipase inhibition. These results allow us to demonstrate the potential of agave bioconjugates, particularly laurate bioconjugates, for the prevention of diseases associated with metabolic syndrome.

Metabolic syndrome (MS) is a group of abnormalities in which obesity, insulin resistance (IR), oxidative stress, and dyslipidemia stand out. This pathology predisposes to the development of cardiovascular diseases and diabetes. The ingestion of linear fructooligosaccharides (FOS) such as inulin reduces conditions such as hyperinsulinemia, increased body fat, and triglyceridemia. When FOS are esterified with fatty acids, they present emulsifying and surfactant properties; however, there are no reports of their function at the biological level. The purpose of this investigation was to evaluate the effect of Agave tequilana Weber's FOS (AtW-FOS) and FOS esterified with lauric acid (FOS-LA) in MS markers in a rat model induced by a HFHC diet. Supplementation with AtW-FOS and FOS-LA decreased IR, improved glucose tolerance, reduced liver weight (19%), plasma triglycerides (24%), and blood pressure (16%) when compared with the untreated MS group. In conclusion, the ingestion of AtW-FOS and FOS-LA has beneficial effects in the prevention of MS alterations, showing a high potential for their application in functional foods.

Fructan fatty acid esters are amphiphilic compounds with potential applications in the food industry due to their surface-active properties on the air/water interface. In this work, the regioselective synthesis of fatty acid esters of short-chain inulin-type fructooligosaccharides (FOS) was carried out in a two-step process comprising sequential enzymatic reactions of transfructosylation and acylation. In the first step, Schedonorus arundinaceus sucrose:sucrose 1-fructosyltransferase (1-SST, EC 2.4.1.99) converted sucrose (600 g/L) into the trisaccharide 1-kestose and the tetrasaccharide nystose in a ratio 9:1 with their sum accounting for 53% (w/w) of total carbohydrates. In the second step, the FOS mixture was transesterified with vinyl laurate in 2-methyl-2-butanol by immobilized Candida antarctica lipase B (CALB) (EC.3.1.1.3). NMR analysis of the synthesized monolaurate FOS esters revealed a regioselective acylation of the 6-OH of the terminal fructosyl moiety of both 1-kestose and nystose. The hydrophilic-lipophilic balance (HLB) values of 6''-O-lauroyl-1-kestose and 6'''-O-lauroylnystose were 14.7 and 15.7, respectively; which suggests their use as oil in water (O/W) emulsifiers. The main compound 6''-O-lauroyl-1-kestose with critical micelle concentration (CMC) of 0.6 mM and surface tension of 45.8 mN/m proved to be a more efficient surfactant than 6'''-O-lauroylnystose (CMC 5.38 mM) and surface tension 36.26 mN/m). We report a regioselective method for the synthesis of lauryl-FOS using the abundant and renewable resource sucrose as the starting substrate.

Yeasts are microscopic fungi inhabiting all Earth environments, including those inhospitable for most life forms, considered extreme environments. According to their habitats, yeasts could be extremotolerant or extremophiles. Some are polyextremophiles, depending on their growth capacity, tolerance, and survival in the face of their habitat's physical and chemical constitution. The extreme yeasts are relevant for the industrial production of value-added compounds, such as biofuels, lipids, carotenoids, recombinant proteins, enzymes, among others. This review calls attention to the importance of yeasts inhabiting extreme environments, including metabolic and adaptive aspects to tolerate conditions of cold, heat, water availability, pH, salinity, osmolarity, UV radiation, and metal toxicity, which are relevant for biotechnological applications. We explore the habitats of extreme yeasts, highlighting key species, physiology, adaptations, and molecular identification. Finally, we summarize several findings related to the industrially-important extremophilic yeasts and describe current trends in biotechnological applications that will impact the bioeconomy.

Lipases are efficient enzymes with promising applications in the nutraceutical and food industry, as they can offer high yields, pure products under achievable reaction conditions, and are an environmentally friendly option. This review addresses the production of high-value-added compounds such as fatty acid esters, with the potential to be used as flavoring agents or antioxidant and antimicrobial agents, as well as structured lipids that offer specific functional properties that do not exist in nature, with important applications in different food products, and pharmaceuticals. In addition, the most recent successful cases of reactions with lipases to produce modified compounds for food and nutraceuticals are reported.

Phlorizin is a low soluble dihydrochalcone with relevant pharmacological properties. In this study, enzymatic fructosylation was approached to enhance the water solubility of phlorizin, and consequently its bioavailability. Three enzymes were assayed for phlorizin fructosylation in aqueous reactions using sucrose as fructosyl donor. Levansucrase (EC 2.4.1.10) from Gluconacetobacter diazotrophicus (Gd_LsdA) was 6.5-fold more efficient than invertase (EC 3.2.1.26) from Rhodotorula mucilaginosa (Rh_Inv), while sucrose:sucrose 1-fructosyltransferase (EC 2.4.1.99) from Schedonorus arundinaceus (Sa_1-SST) failed to modify the non-sugar acceptor. Gd_LsdA synthesized series of phlorizin mono- di- and tri-fructosides with maximal conversion efficiency of 73 %. The three most abundant products were identified by ESI-MS and NMR analysis as beta-D-fructofuranosyl-(2 -> 6)-phlorizin (P1a), phlorizin-4'-O-beta-D-fructofuranosyl-(2 -> 6)-D-fructofuranoside (P2c) and phlorizin-4-O-monofructofuranoside (P1b), respectively. Purified P1a was 16 times (30.57 g L-1 at 25 degrees C) more soluble in water than natural phlorizin (1.93 g L-1 at 25 degrees C) and exhibited 44.56 % free radical scavenging activity. Gd_LsdA is an attractive candidate enzyme for the scaled synthesis of phlorizin fructosides in the absence of co-solvent.

The dry washing method is an alternative to replace water washing, thereby reducing the negative impacts of contamination. However, commercial adsorbents come from industrial processes that, due to their composition, may not be such a sustainable resource in the global biodiesel production process. In this study, the use of organic residues, such as sawdust, coconut fiber, nutshell, rice husk, and water hyacinth fiber, were proposed as bioadsorbents for the purification of biodiesel from waste cooking oil. Quality parameters such as the acid number, water content, and free and total glycerin content were evaluated and compared with those after purification with commercial adsorbents (Magnesol and Amberlite BD10DRY). Promising results were obtained using sawdust in the purification process, achieving a reduction in the acid number value of 31.3% respect to the unpurified biodiesel. Indeed, the reduction with sawdust was more efficient than with Amberlite BD10DRY (that increased the acid number). In addition, sawdust reduced free glycerin by 54.8%, again more efficient than Amberlite BD10DRY. The total glycerin values were similar between commercial adsorbents and sawdust. Water content after purification with sawdust was similar to the obtained with Amberlite BD10DRY and better than with Magnesol (399, 417, and 663 mg/kg respectively). These results show that sawdust can be used as an alternative bioadsorbent in a dry purification method for biodiesel, generating less environmental impact.

In this study, the antifungal activity of AgNPs was tested against C. tropicalis (pathogen fungi) and S. boulardii (probiotic). The effectiveness of the AgNPs was assessed by comparing their antifungal activity with a triazole antifungal drug fluconazole and amphotericin B. The AgNPs have a polygonal-like shape (average size of 35 +/- 15 nm) with 1.2% wt. of metallic silver stabilized with 18.8% wt. of polyvinylpyrrolidone (PVP) in 80% wt. of distilled water. The results revealed that 35 mu g/mL of fluconazole inhibits 55-60% of both fungal cell growth. As for amphotericin B, 5 mu g/mL is sufficient to inhibit more than 95% of both fungal cells. For AgNPs, 25 mu g/mL was needed to inhibit 90% of the C. tropicalis cell growth, but remarkably, 50% of the S. boulardii cell population remains viable, which can potentiate cell reproduction. Our results could initiate the development of AgNPs possessing selective specificity against pathogenic fungal species.