Levan Production by Suhomyces kilbournensis Using Sugarcane Molasses as a Carbon Source in Submerged Fermentation.

The valorization of byproducts from the sugarcane industry represents a potential alternative method with a low energy cost for the production of metabolites that are of commercial and industrial interest. The production of exopolysaccharides (EPSs) was carried out using the yeast Suhomyces kilbournensis isolated from agro-industrial sugarcane, and the products and byproducts of this agro-industrial sugarcane were used as carbon sources for their recovery. The effect of pH, temperature, and carbon and nitrogen sources and their concentration in EPS production by submerged fermentation (SmF) was studied in 170 mL glass containers of uniform geometry at 30 °C with an initial pH of 6.5. The resulting EPSs were characterized with Fourier-transform infrared spectroscopy (FT-IR). The results showed that the highest EPS production yields were 4.26 and 44.33 g/L after 6 h of fermentation using sucrose and molasses as carbon sources, respectively. Finally, an FT-IR analysis of the EPSs produced by S. kilbournensis corresponded to levan, corroborating its origin. It is important to mention that this is the first work that reports the production of levan using this yeast. This is relevant because, currently, most studies are focused on the use of recombinant and genetically modified microorganisms; in this scenario, Suhomyces kilbournensis is a native yeast isolated from the sugar production process, giving it a great advantage in the incorporation of carbon sources into their metabolic processes in order to produce levan sucrose, which uses fructose to polymerize levan.

Evaluation of the Biological Properties of an Optimized Extract of Polygonum cuspidatum Using Ultrasonic-Assisted Extraction.

Phytochemicals are natural compounds found in plants that have potential health benefits such as antioxidants, anti-inflammatory and anti-cancer properties, and immune reinforcement. Polygonum cuspidatum Sieb. et Zucc. is a source rich in resveratrol, traditionally consumed as an infusion. In this study, P. cuspidatum root extraction conditions were optimized to increase antioxidant capacity (DPPH, ABTS+), extraction yield, resveratrol concentration, and total polyphenolic compounds (TPC) via ultrasonic-assisted extraction using a Box-Behnken design (BBD). The biological activities of the optimized extract and the infusion were compared. The optimized extract was obtained using a solvent/root powder ratio of 4, 60% ethanol concentration, and 60% ultrasonic power. The optimized extract showed higher biological activities than the infusion. The optimized extract contained 16.6 mg mL-1 resveratrol, high antioxidant activities (135.1 µg TE mL-1 for DPPH, and 230.4 µg TE mL-1 for ABTS+), TPC (33.2 mg GAE mL-1), and extraction yield of 12.4%. The EC50 value (effective concentration 50) of the optimized extract was 0.194 µg mL-1, which revealed high cytotoxic activity against the Caco-2 cell line. The optimized extract could be used to develop functional beverages with high antioxidant capacity, antioxidants for edible oils, functional foods, and cosmetics.

Physicochemical, Mechanical, and Structural Properties of Bio-Active Films Based on Biological-Chemical Chitosan, a Novel Ramon (Brosimum alicastrum) Starch, and Quercetin.

The properties of biological-chemical chitosan (BCh) films from marine-industrial waste and a non-conventional Ramon starch (RS) (Brosimum alicastrum) were investigated. Blended films of BCh/RS were prepared to a volume ratio of 4:1 and 1:4, named (BChRS-80+q, biological-chemical chitosan 80% v/v and Ramon starch, BChRS-20+q, biological-chemical chitosan 20% v/v and Ramon starch, both with quercetin), Films from commercial chitosan (CCh) and corn starch (CS), alone or blended (CChCS-80+q, commercial chitosan 80% v/v and corn starch, CChCS-20+q commercial chitosan 20% v/v and corn starch, both with quercetin) were also prepared for comparison purposes. Films were investigated for their physicochemical characteristics such as thickness, moisture, swelling, water-vapor permeability, and water solubility. In addition, their mechanical and structural properties were studied using Fourier Transform Infrared Spectroscopy (FTIR), Thermogravimetric analysis (TGA) and Scanning Electron Microscopy (SEM) techniques. Antioxidant activity was evaluated as radical scavenging, and antimicrobial effect was also determined. The BCh and RS films presented similar tensile strength values compared with commercial biopolymers. Only films with chitosan presented antioxidant and antimicrobial activity. The FTIR spectra confirmed the interactions between functional groups of the biopolymers. Although, BChRS-80+q and BChRS-20+q films exhibited poor mechanical performance compared to their commercial counterparts, they showed good thermal stability, and improved antioxidant and antimicrobial activity in the presence of quercetin. BChRS-80+q and BChRS-20+q films have promising applications due to their biological activity and mechanical properties, based on a novel material that has been underutilized (Ramon starch) that does not compete with materials for human feeding and may be used as a coating for food products.

Physicochemical composition, phytochemical analysis and biological activity of ciricote (Cordia dodecandra A. D.C.) fruit from Yucatán.

The physicochemical properties, proximate composition, minerals, total polyphenols, carotenoids, phenolic compounds, antioxidant, and antibacterial activities of ciricote (Cordia dodecandra A. DC.) tropical fruit were investigated. Minerals were quantified by using micro-Energy Dispersive X-Ray Fluorescence. Lutein and ß-carotene were identified in ciricote fruit by using UPLC-PDA analysis. The highest values of the total polyphenols content and antioxidant activity were presented in ethanolic crude extracts obtaining by the ultrasonic-assisted method with freeze-dried fruit. The phenolic acids profile was identified and quantified by UPLC-PDA-ESI-MS. The main phenolic acids were caffeoyl hexoside, rufescenolide, quercetin 3-O-rutinoside, and rosmarinic acid. The ciricote extracts presented antibacterial activity against Staphylococus aureus (Gram+) and Salmonella typhymurium (Gram-). In conclusion, the ciricote (Cordia dodecandra A. DC.) tropical fruits could be very useful source of biological macromolecules, micro-elements, and phytochemical compounds for the food and pharmaceutical industry.

Optimization of the Biodegradation of Aliphatic, Aromatic, and UCM Hydrocarbons from Light Crude Oil in Marine Sediment Using Response Surface Methodology (RSM).

This study describes the optimization of the biodegradation of total aliphatic (tAHCs), total aromatic (tPAHs), and unresolved complex mixture (UCM) hydrocarbons from light crude oil in marine sediment. The response surface methodology (RSM), with a Box-Behnken design, was used to optimize the hydrocarbon fraction degradation, reported as degradation efficiency (E (%)), using four independent variables (inoculum, dispersant, light oil concentration, and carbon/nitrogen ratio), all at three levels. Analysis of variance (ANOVA) showed R2 values of 0.976, 0.974, and 0.975 for tAHCs, tPAHs, and UCM, respectively. All fractions exhibited a statistically significant effect (P < 0.05) in the second-order quadratic regression model for degradation. According to the models, the optimal degradation prediction was: 81.03% for tAHCs, 85.96% for tPAHs, and 92.86% for UCM. This work highlights the possibility of carrying out efficient biodegradation, of more than 80%, through an optimization process using different light oil concentrations, opening up possibilities of multiple response optimization.

Zinc Oxide and Copper Chitosan Composite Films with Antimicrobial Activity.

The role of the oral microbiome and its effect on dental diseases is gaining interest. Therefore, it has been sought to decrease the bacterial load to fight oral cavity diseases. In this study, composite materials based on chitosan, chitosan crosslinked with glutaraldehyde, chitosan with zinc oxide particles, and chitosan with copper nanoparticles were prepared in the form of thin films, to evaluate a new alternative with a more significant impact on the oral cavity bacteria. The chemical structures and physical properties of the films were characterized using by Fourier transform infrared spectroscopy (FTIR,) Raman spectroscopy, X-ray photoelectron spectroscopy (XPS), elemental analysis (EDX), thermogravimetric analysis (TGA), X-ray diffraction (XRD), scanning electron microscopy (SEM), and contact angle measurements. Subsequently, the antimicrobial activity of each material was evaluated by agar diffusion tests. No differences were found in the hydrophilicity of the films with the incorporation of ZnO or copper particles. Antimicrobial activity was found against S. aureus in the chitosan film crosslinked with glutaraldehyde, but not in the other compositions. In contrast antimicrobial activity against S. typhimurium was found in all films. Based on the data of present investigation, chitosan composite films could be an option for the control of microorganisms with potential applications in various fields, such as medical and food industry.

Kinetic, Thermodynamic, Physicochemical, and Economical Characterization of Pectin from Mangifera indica L. cv. Haden Residues.

The effect of temperature (60, 70, 80, and 90 °C) and time (30, 45, 60, 75, and 90 min) on citric acid extraction of Haden mango (Mangifera indica L. cv. Haden) peel pectin was evaluated in the present study. In order to obtain a better understanding of both the extraction process and the characteristics of the pectin (obtained from an agro-industrial waste) for a future scaling process, the following characterizations were performed: (1) Kinetic, with the maximum extraction times and yields at all evaluated temperatures; (2) thermodynamic, obtaining activation energies, enthalpies, entropies, and Gibbs free energies for each stage of the process; (3) physicochemical (chemical analysis, monosaccharide composition, degree of esterification, galacturonic acid content, free acidity, Fourier-transform infrared spectroscopy, thermogravimetric and derivative thermogravimetric analyses); and (4) economical, of the pectin with the highest yield. The Haden mango peel pectin was found to be characterized by a high-esterified degree (81.81 ± 0.00%), regular galacturonic acid content (71.57 ± 1.26%), low protein (0.83 ± 0.05%) and high ash (3.53 ± 0.02%) content, low mean viscometric molecular weight (55.91 kDa), and high equivalent weight (3657.55 ± 8.41), which makes it potentially useful for food applications.

Physicochemical and Optical Characterization of Citrus aurantium Derived Biochar for Solar Absorber Applications.

Agro-industrial waste valorization is an attractive approach that offers new alternatives to deal with shrinkage and residue problems. One of these approaches is the synthesis of advanced carbon materials. Current research has shown that citrus waste, mainly orange peel, can be a precursor for the synthesis of high-quality carbon materials for chemical adsorption and energy storage applications. A recent approach to the utilization of advanced carbon materials based on lignocellulosic biomass is their use in solar absorber coatings for solar-thermal applications. This study focused on the production of biochar from Citrus aurantium orange peel by a pyrolysis process at different temperatures. Biochars were characterized by SEM, elemental analysis, TGA-DSC, FTIR, DRX, Raman, and XPS spectroscopies. Optical properties such as diffuse reflectance in the UV-VIS-NIR region was also determined. Physical-chemical characterization revealed that the pyrolysis temperature had a negative effect in yield of biochars, whereas biochars with a higher carbon content, aromaticity, thermal stability, and structural order were produced as the temperature increased. Diffuse reflectance measurements revealed that it is possible to reduce the reflectance of the material by controlling its pyrolysis temperature, producing a material with physicochemical and optical properties that could be attractive for use as a pigment in solar absorber coatings.

Effect of solvent polarity on the Ultrasound Assisted extraction and antioxidant activity of phenolic compounds from habanero pepper leaves (Capsicum chinense) and its identification by UPLC-PDA-ESI-MS/MS.

Phenolic compounds are secondary metabolites involved in plant adaptation processes. The development of extraction procedures, quantification, and identification of this compounds in habanero pepper (Capsicum chinense) leaves can provide information about their accumulation and possible biological function. The main objective of this work was to study the effect of the UAE method and the polarity of different extraction solvents on the recovery of phenolic compounds from C. chinense leaves. Quantification of the total phenolic content (TPC), antioxidant activity (AA) by ABTS+ and DPPH radical inhibition methods, and the relation between the dielectric constant (ε) as polarity parameter of the solvents and TPC using Weibull and Gaussian distribution models was analyzed. The major phenolic compounds in C. chinense leaves extracts were identified and quantified by UPLC-PDA-ESI-MS/MS. The highest recovery of TPC (24.39 ± 2.41 mg GAE g-1 dry wt) was obtained using MeOH (50%) by UAE method. Correlations between TPC and AA of 0.89 and 0.91 were found for both radical inhibition methods (ABTS+ and DPPH). The Weibull and Gaussian models showed high regression values (0.93 to 0.95) suggesting that the highest phenolic compounds recovery is obtained using solvents with "ε" values between 35 and 52 by UAE. The major compounds were identified as N-caffeoyl putrescine, apigenin, luteolin and diosmetin derivatives. The models presented are proposed as a useful tool to predict the appropriate solvent composition for the extraction of phenolic compounds from C. chinense leaves by UAE based on the "ε" of the solvents for future metabolomic studies.

Ultrasound-Assisted Extraction Optimization of Phenolic Compounds from Citrus latifolia Waste for Chitosan Bioactive Nanoparticles Development.

Bioactive Phenols-loaded chitosan nanoparticles (PL-CNps) were developed by ionic gelation from Persian lemon (Citrus latifolia) waste (PLW) and chitosan nanoparticles. Response Surface Methodology (RSM) was used to determine the optimal Ultrasound-Assisted Extraction (UAE) conditions for the total phenolic compounds (TPC) recovery from PLW (58.13 mg GAE/g dw), evaluating the ethanol concentration, extraction time, amplitude, and solid/liquid ratio. Eight compounds expressed as mg/g dry weight (dw) were identified by ultra-performance liquid chromatography coupled photo diode array (UPLC-PDA) analysis: eriocitrin (20.71 ± 0.09), diosmin (18.59 ± 0.13), hesperidin (7.30 ± 0.04), sinapic acid (3.67 ± 0.04), catechin (2.92 ± 0.05), coumaric acid (2.86 ± 0.01), neohesperidin (1.63 ± 0.00), and naringenin (0.44 ± 0.00). The PL-CNps presented size of 232.7 nm, polydispersity index of 0.182, Z potential of -3.8 mV, and encapsulation efficiency of 81.16%. The results indicated that a synergic effect between phenolic compounds from PLW and chitosan nanoparticles was observed in antioxidant and antibacterial activity, according to Limpel's equation. Such results indicate that PLW in such bioprocesses shows excellent potential as substrates for the production of value-added compounds with a special application for the food industry.

Effect of bio-chemical chitosan and gallic acid into rheology and physicochemical properties of ternary edible films.

The ternary edible films based on commercial or bio-chemical chitosan (CCh or BCh) and starch (S) with different concentration of gallic acid (GA) were produced by casting. This work analyzed the effect of different type of Ch and GA concentrations (1.25 mg/gTB and 2.5 mg/gTB) on the rheological properties of the film-forming solutions, as well as physicochemical (thickness, moisture content, swelling index, water vapor permeability (WVP), tensile strength, microstructure, opacity, and color) and antioxidant properties of the edible films themselves. The chemical cross-linking between the GA and Ch into the edible films of biopolymers could form of ester linkages between the carboxyl groups of GA and the hydroxyl groups of Ch. Results showed that the ternary edible films of BCh-S-GA5.0 resulted with a positive effect on their rheological (η0, λ, and p), physicochemical (WVP, swelling, tensile strength, microstructure) and antioxidant properties. The greatest incorporation of polyphenol into the BCh-S-GA5.0 films greatly reduced WVP and swelling values in approximately 61.01% and 23%, respectively also the tensile strength value increased to 37.372 MPa. The ternary edible films developed in this study showed many desirable characteristics, which could potentially be used as bioactive packaging films for food applications.

Activity of chitin deacetylase from Colletotrichum gloeosporioides on chitinous substrates.

Production of chitin deacetylases from the phytopathogenic fungus Colletotrichum gloeosporioides was successfully achieved by submerged fermentation. The highest specific activity of 0.018 U mg(-1) of protein was obtained after 96 h of cultivation at pH 6 and 28°C. Two bands with molecular weights of 35 kDa and 170 kDa determined with SDS-PAGE displayed deacetylase activities as detected in the zymograms. Reacetylated commercial chitosan (52% acetylation degree) was used as substrate for the extracellular crude extract in order to estimate the kinetic parameters of acetate production as undirected deacetylation measurement. The highest acetate production of 12.8 µmol mL(-1) was obtained using 7.5 mg mL(-1) of substrate. The produced enzyme from C. gloeosporioides achieved up to 25% deacetylation of a chitin substrate (hydrolyzed biological chitin) having 80% degree of acetylation, MW of 102×10(3) g mol(-1) and a crystallinity index of ca. 60%.

Structural characterization of chitin and chitosan obtained by biological and chemical methods.

Chitin production was biologically achieved by lactic acid fermentation (LAF) of shrimp waste (Litopenaeus vannameii) in a packed bed column reactor with maximal percentages of demineralization (D(MIN)) and deproteinization (D(PROT)) after 96 h of 92 and 94%, respectively. This procedure also afforded high free astaxanthin recovery with up to 2400 µg per gram of silage. Chitin product was also obtained from the shrimp waste by a chemical method using acid and alkali for comparison. The biologically obtained chitin (BIO-C) showed higher M(w) (1200 kDa) and crystallinity index (I(CR)) (86%) than the chemically extracted chitin (CH-C). A multistep freeze-pump-thaw (FPT) methodology was applied to obtain medium M(w) chitosan (400 kDa) with degree of acetylation (DA) ca. 10% from BIO-C, which was higher than that from CH-C. Additionally, I(CR) values showed the preservation of crystalline chitin structure in BIO-C derivatives at low DA (40-25%). Moreover, the FPT deacetylation of the attained BIO-C produced chitosans with bloc copolymer structure inherited from a coarse chitin crystalline morphology. Therefore, our LAF method combined with FPT proved to be an affective biological method to avoid excessive depolymerization and loss of crystallinity during chitosan production, which offers new perspective applications for this material.

Effect of temperature on chitin and astaxanthin recoveries from shrimp waste using lactic acid bacteria.

The chitin and astaxanthin recoveries by lactic acid fermentation of shrimp wastes (Litopenaeus sp) were conducted in bed-column reactors at 15, 20, 25, 30, 35, 40 and 45 degrees C. The response surface methodology showed that the fermentations carried out in the 27-36 degrees C temperature range with lactic acid above 0.319 mmol/g resulted in the highest demineralization. The maximal deproteinizations were attained from 30 to 40 degrees C. The extraction of free-astaxanthin did not present significant differences between 20 and 35 degrees C and the proportion of cis-stereoisomer forms increased with temperature. The growth rates of Lactobacillus plantarum were estimated in the 15-45 degrees C range and analyzed by Arrhenius and square root models. The cardinal values were 3.94 and 51.7 degrees C for minimum and maximum temperatures, respectively, with activation energy of 43.38 Jmol(-1).

Evaluation of chitosans and Pichia guillermondii as growth inhibitors of Penicillium digitatum.

Chitosans were obtained by room-temperature-homogeneous-deacetylation (RTHD) and freeze-pump-out-thaw-heterogeneous-deacetylation (FPT) from chitins purified from fermentations. Commercial chitosan was deacetylated by three-FPT-cycles. Chitosans and Pichia guillermondii were evaluated on the growth of Penicillium digitatum. Medium molecular weight (M(W)) chitosans displayed higher inhibitory activity against the yeast than low M(W) biopolymers. Chitosans with low degree of acetylation (DA) were inhibitory for yeast and mould. Therefore, a low M(W) and high DA chitosan was selected for use against moulds combined with yeasts. Biopolymer and yeasts presented an additive effect, since chitosans were effective to delay spore germination, whereas yeast decreased apical fungal growth.

One-solvent extraction of astaxanthin from lactic acid fermented shrimp wastes.

Free astaxanthin one-solvent extractions with ethanol, acetone, and liquid 1,1,1,2-tetrafluoroethane from raw and lactic acid fermented (ensilaged) shrimp residues were investigated. The total carotenoid recovery from ensilaged shrimp wastes was higher than that from non-ensilaged ones as assessed by HPLC analyses. Acetone gave the highest extraction yields of free astaxanthin with up to 115 microg/g of material. Moreover, liquid tetrafluoroethane is reported for the first time in a successful one-solvent extraction of carotenoids from shrimp.