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Food Chemistry Jan 2022In this work, magnetic chitosan (MCH) beads were synthesized by phase-inversion method, and grafted with polydopamine (PDA) and then used for direct immobilization of...
In this work, magnetic chitosan (MCH) beads were synthesized by phase-inversion method, and grafted with polydopamine (PDA) and then used for direct immobilization of Candida rugosa lipase by Schiff base reaction. The amount of immobilized enzyme and the retained activity were found to be 47.3 mg/g and 72.8%, respectively, at pH 7.0, and at 25 °C. The apparent Km (9.7 mmol/L), and Vmax (384 U/mg) values of the immobilized lipase were significantly changed compared to the free lipase. The MCH@PDA-lipase was better thermal and storage stability at different temperatures than those of the free lipase. In hexane medium, the esterification reaction results showed that the maximum conversions of isoamylalcohol and isopentyl alcohol to isoamyl acetate and isopentyl acetate using the MCH@PDA-lipase were found to be 98.4 ± 1.3% and 73.7 ± 0.7%, respectively. These results showed that the MCH@PDA-lipase can be used as an operative immobilized enzyme system for many biotechnological applications.
Topics: Chitosan; Enzyme Stability; Enzymes, Immobilized; Esters; Hydrogen-Ion Concentration; Lipase; Magnetic Phenomena; Saccharomycetales
PubMed: 34348221
DOI: 10.1016/j.foodchem.2021.130699 -
Journal of the Science of Food and... Apr 2022Black soldier fly larvae (BSFL) offer very good prospects for the production of cheap and high-quality dietary protein. This insect is able to consume low-quality...
BACKGROUND
Black soldier fly larvae (BSFL) offer very good prospects for the production of cheap and high-quality dietary protein. This insect is able to consume low-quality substrates, including food waste. The properties and safety of the compost obtained are largely determined by its microbiome. However, while the bacterial component of the BSFL-compost microbiome has been much studied, little is known about its fungal component. In plant-based rearing substrates both the biomass and the metabolic activity of fungi often exceed the biomass and the activity of prokaryotes. So, the purpose of this study was to investigate the fungal community of the compost produced by BSF larvae reared on a food waste substrate.
RESULTS
Community structure was determined by metabarcoding of the ITS (internal transcribed spacer) region. Species composition and abundance were determined using the plating technique and subsequent identification of the isolated pure cultures. It was found that the primary mycobiome of the food waste substrate that was used consisted of 19 families, represented mainly by phytopathogenic and endophytic genera. Larva incubation led to the complete elimination of all mycelial fungi from the resulting compost. The final mycobiome consisted of only two yeast species, Pichia kudriavzevii and Diutina rugosa, with a total abundance of 1.2 × 10 CFU g .
CONCLUSIONS
The rearing of BSFL led to the complete elimination of mycelial fungi from its feed substrate. The final compost lacked harmful fungi, including molds. This information may be crucial for BSF compost utilization. The phenomenon was also an interesting aspect of zoomicrobial interactions in nature and agriculture. © 2021 Society of Chemical Industry.
Topics: Animals; Composting; Diptera; Food; Larva; Mycobiome; Refuse Disposal
PubMed: 34687463
DOI: 10.1002/jsfa.11601 -
Probiotics and Antimicrobial Proteins Feb 2024Probiotic microorganisms are incorporated in foods due to their numerous health benefits. We investigated lactic acid bacteria (LAB) and yeasts isolated from goat milk...
Probiotic microorganisms are incorporated in foods due to their numerous health benefits. We investigated lactic acid bacteria (LAB) and yeasts isolated from goat milk in Nigeria for novel probiotic strains. In this study, a total of 27 LAB and 23 yeast strains were assessed for their probiotic potentials. Only six LAB strains (Weissella cibaria GM 93m3, Weissella confusa GM 92m1, Pediococcus acidilactici GM 18a, Pediococcus pentosaceus GM 23d, Lactiplantibacillus pentosus GM 102s4, Limosilactobacillus fermentum GM 30m1) and four yeast strains (Candida tropicalis 12a, C. tropicalis 33d, Diutina rugosa 53b, and D. rugosa 77a) identified using partial 16S and 26S rDNA sequencing, respectively, showed survival at pH 2.5, 0.3% bile salt, and simulated gastrointestinal conditions and possessed auto-aggregative and hydrophobic properties, thus satisfying key in vitro criteria as probiotics. All LAB strains showed coaggregation properties and antimicrobial activities against pathogens. Pediococcus pentosaceus GM 23d recorded the strongest coaggregation percentage (34-94%) against 14 pathogens, while W. cibaria GM 93m3 showed the least (6-57%) against eight of the 14 pathogens. The whole cell and extracellular extracts of LAB and yeast strains, with the exception of D. rugosa 77a, had either 2,2-diphenyl-1-picryl-hydrazyl and/or hydroxyl radical scavenging activity. In conclusion, all six LAB and four yeast strains are important probiotic candidates that can be further investigated for use as functional starter cultures.
Topics: Animals; Lactobacillales; Milk; Nigeria; Goats; Probiotics; Yeasts; Pediococcus pentosaceus
PubMed: 36520357
DOI: 10.1007/s12602-022-10022-w -
Journal of Biotechnology Jul 2023Metal-organic frameworks (MOFs) are used as ideal support materials thanks to their unique properties and have become the focus of interest in enzyme immobilization...
Metal-organic frameworks (MOFs) are used as ideal support materials thanks to their unique properties and have become the focus of interest in enzyme immobilization studies, especially in recent years. In order to increase the catalytic activity and stability of Candida rugosa lipase (CRL), a new fluorescence-based MOF (UiO-66-Nap) derived from UiO-66 was synthesized. The structures of the materials were confirmed by spectroscopic techniques such as FTIR, H NMR, SEM, and PXRD. CRL was immobilized on UiO-66-NH and UiO-66-Nap by adsorption technique and immobilization and stability parameters of UiO-66-Nap@CRL were examined. Immobilized lipases UiO-66-Nap@CRL exhibited higher catalytic activity (204 U/g) than UiO-66-NH @CRL (168 U/g), which indicates that the immobilized lipase (UiO-66-Nap@CRL) carries sulfonate groups, this is due to strong ionic interactions between the surfactant's polar groups and certain charged locations on the protein surface. The Free CRL lost its catalytic activity completely at 60 °C after 100 min, while UiO-66-NH @CRL and UiO-66-Nap@CRL retained 45 % and 56 % of their catalytic activity at the end of 120 min, respectively. After 5 cycles, the activity of UiO-66-Nap@CRL remained 50 %, while the activity of UiO-66-NH @CRL was about 40 %. This difference is due to the surfactant groups (Nap) in UiO-66-Nap@CRL. These results show that the newly synthesized fluorescence-based MOF derivative (UiO-66-Nap) can be an ideal support material for enzyme immobilization and can be used successfully to protect and increase the activities of enzymes.
Topics: Biocatalysis; Metal-Organic Frameworks; Surface-Active Agents; Candida; Enzymes, Immobilized; Lipase
PubMed: 37301292
DOI: 10.1016/j.jbiotec.2023.06.003 -
Biodegradation Oct 2021In this work, strains of Bacillus subtilis were inoculated in consortium with Rhodotorula mucilaginosa into spent soy oil as aiming to biological treatment and low-cost...
In this work, strains of Bacillus subtilis were inoculated in consortium with Rhodotorula mucilaginosa into spent soy oil as aiming to biological treatment and low-cost reuse. The microorganisms were previously isolated and selected for the lipolytic capacity of the alperujo residue generated during the processing of olive oil. For fermentation, bioassays containing Rhodotorula mucilaginosa isolated from alperujo and Candida rugosa CCMA 00371, both co-inoculated with Bacillus subtilis CCMA 0085 in medium containing (% w/v) 0.075 glucose and 0.375 (NH) PO in 75 mL of water and 75 mL of spent soy oil. Despite the low biomass productivity, it has favorable characteristics to be used in animal feed supplementation. Spent soy oil was used as a carbon source proven by Bartha respirometer. The strains of R. mucilaginosa UFLA RAS 144 and B. subtilis CCMA 0085 are promising inoculants for oil degradation and can be applied in a waste treatment system.
Topics: Biodegradation, Environmental; Hydrolysis; Microbial Consortia; Rhodotorula; Saccharomycetales; Soybean Oil; Technology
PubMed: 34046776
DOI: 10.1007/s10532-021-09951-0 -
Biotechnology and Applied Biochemistry Jun 2021The present study aimed at preparing three biocatalysts via physical adsorption of lipases from Candida rugosa (CRL), Mucor javanicus, and Candida sp. on a hydrophobic...
The present study aimed at preparing three biocatalysts via physical adsorption of lipases from Candida rugosa (CRL), Mucor javanicus, and Candida sp. on a hydrophobic and mesoporous support (Diaion HP-20). These biocatalysts were later applied to the synthesis of aromatic esters of apple peel and citrus (hexyl butyrate), apple and rose (geranyl butyrate), and apricot and pineapple (propyl butyrate). Scanning electron microscopy and gel electrophoresis confirmed a selective adsorption of lipases on Diaion, thus endorsing simultaneous immobilization and purification. Gibbs free energy (∆G) evinced the spontaneity of the process (-17.9 kJ/mol ≤ ∆G ≤ -5.1 kJ/mol). Maximum immobilized protein concentration of 30 mg/g support by CRL. This biocatalyst was the most active in olive oil hydrolysis (hydrolytic activity of 126.0 ± 2.0 U/g) and in the synthesis of aromatic esters. Maximum conversion yield of 89.1% was attained after 150 Min for the synthesis of hexyl butyrate, followed by the synthesis of geranyl butyrate (87.3% after 240 Min) and propyl butyrate (80.0% after 150 Min). CRL immobilized on Diaion retained around 93% of its original activity after six consecutive cycles of 150 Min for the synthesis of hexyl butyrate.
Topics: Enzymes, Immobilized; Esters; Hydrocarbons, Aromatic; Hydrophobic and Hydrophilic Interactions; Lipase; Mucor; Particle Size; Saccharomycetales; Surface Properties
PubMed: 32438471
DOI: 10.1002/bab.1959 -
Microbiome Sep 2023Changes in population heterozygosity and genetic diversity play important roles in mediating life history traits of organisms; these changes often lead to phenotypic...
BACKGROUND
Changes in population heterozygosity and genetic diversity play important roles in mediating life history traits of organisms; these changes often lead to phenotypic evolution in offspring, which become superior to their parents. In the present study, we examined phenotypic differentiation, the intestinal microbiome composition, and metabolism shift in the oriental fruit fly (Bactrocera dorsalis) by comparing an inbred (monophyletic) original population and an outbred (mixed) invasive population.
RESULTS
The results showed that the outbred population of B. dorsalis had significantly higher biomass, adult longevity, and fecundity than the inbred population. Additionally, intestinal microflora analysis revealed that both Diutina rugosa and Komagataeibacter saccharivorans were significantly enriched in the outbred population with higher genetic heterozygosity. D. rugosa enrichment altered amino acid metabolism in the intestinal tract, and supplementing essential amino acids (e.g. histidine and glutamine) in the diet led to an increase in pupal weight of the outbred population. Additionally, transcriptome analysis revealed that the HSPA1S gene was significantly downregulated in the outbred population. HSPA1S was involved in activation of the JNK-MAPK pathway through negative regulation, caused the upregulation of juvenile hormone (JH), and led to an increase in biomass in the outbred flies.
CONCLUSION
In conclusion, the outbred population had an altered intestinal microbe composition, mediating metabolism and transcriptional regulation, leading to phenotypic differentiation; this may be a potential mechanism driving the global invasion of B. dorsalis. Thus, multiple introductions could lead to invasiveness enhancement in B. dorsalis through population mixing, providing preliminary evidence that changes in the intestinal microbiome can promote biological invasion. Video Abstract.
Topics: Animals; Gastrointestinal Microbiome; Drosophila; Tephritidae; Gene Expression Profiling; Gene Expression Regulation
PubMed: 37759251
DOI: 10.1186/s40168-023-01664-1 -
Biomedicine & Pharmacotherapy =... Oct 2020Diet and commercially available supplements can significantly impact the gut microbial composition; however, the effects of supplements often lack scientific data...
Diet and commercially available supplements can significantly impact the gut microbial composition; however, the effects of supplements often lack scientific data demonstrating the effects on healthy and diseased individuals. Hence, it was investigated, whether a frequently used supplement in humans, Candida rugosa lipase (CRL), gets delivered active beyond the stomach in the intestinal tract of C57BL/6 J mice and its impact on the gut microbial community and environment. We showed for the first time the movement of CRL in an active state through the mouse digestive tract by determination of intestinal CRL activity and free fatty acids concentrations. The short- and long-term administration of CRL resulted in significant alterations of the gut microbiome, favoring the growth of, for instance, Verrucomicrobia but also other species associated with normal body mass index (BMI) or butyrate expression, both considered beneficial. In addition, we showed that these changes persisted after supplementation and that gut barrier integrity was unaffected by the treatment. In conclusion, CRL can be delivered in an active state beyond the stomach and supplementation altered the murine gut microbiome favoring beneficial bacterial species, which may be of relevance in humans in healthy but also potentially in disease states.
Topics: Animals; Bacteria; Gastrointestinal Microbiome; Gastrointestinal Tract; Lipase; Mice, Inbred C57BL; RNA, Ribosomal, 16S; Saccharomycetales
PubMed: 32771891
DOI: 10.1016/j.biopha.2020.110579 -
International Journal of Biological... May 2021In this paper, an epoxy-activated cloisite (ECL) was prepared as a new heterofunctional carrier via a reaction between cloisite 30B (CL) and epichlorohydrin and utilized...
Covalent immobilization of lipase from Candida rugosa on epoxy-activated cloisite 30B as a new heterofunctional carrier and its application in the synthesis of banana flavor and production of biodiesel.
In this paper, an epoxy-activated cloisite (ECL) was prepared as a new heterofunctional carrier via a reaction between cloisite 30B (CL) and epichlorohydrin and utilized for covalent immobilization of lipase from Candida rugosa. The lipase immobilized on the ECL (LECL) was successfully used in the olive oil hydrolysis, synthesis of isoamyl acetate (banana flavor), and biodiesel production. The TGA, FT-IR, SEM, and XRD were used to characterize CL, ECL, and LECL. The influences of temperature, pH, thermal stability, and storage capacity were examined in the olive oil hydrolysis. The effects of solvent, temperature, time, water content, and substrates molar ratio on the yields of ester and biodiesel were also investigated. In the optimized conditions, the hydrolytic activity of LECL was 1.85 ± 0.05 U/ mg, and the maximum yield of ester and biodiesel was 91.6% and 95.4%, respectively. The LECL showed good thermal stability and storage capacity compared to the free lipase. Additionally, LECL was reusable for both esterification and transesterification after being used for nine cycles.
Topics: Biofuels; Enzymes, Immobilized; Esterification; Flavoring Agents; Fungal Proteins; Lipase; Saccharomycetales
PubMed: 33667558
DOI: 10.1016/j.ijbiomac.2021.02.146 -
Bioprocess and Biosystems Engineering Nov 2020The active site of Candida rugosa lipase (CRL) is mainly hydrophilic on its external face and hydrophobic on the internal side, and calix[n]arene-based surfactants form...
The active site of Candida rugosa lipase (CRL) is mainly hydrophilic on its external face and hydrophobic on the internal side, and calix[n]arene-based surfactants form complexes with protein residues or with strong hydrogen bonds to open up the lid. Therefore, the activity of lipase persists for a long time. In this work, a series of cyclic and acyclic anionic surfactants (sodium dodecyl sulfate (SDS), p-sulfonatocalix[4]arene, and p-sulfonatocalix[8]arene) and zwitterionic surfactants (L-proline and L-proline derivative of calix[4]arene) were used to examine the relationship between the surfactants' molecular structures and their effects on the hydrolytic activity of CRL. We explored the effects of different surfactant concentrations, ring effects, and mixing times on CRL activity and several kinetic parameters. The results demonstrated that cyclic compounds were more effective than linear structures for increasing CRL activity and the highest enzyme activity was obtained by the addition of the calix[4]-L-proline derivative. This zwitterionic compound (calix[4]-L-proline derivative) maintains the active center of enzyme and conformation by enabling electrostatic interactions and hydrogen bonding with both the acidic and basic amino acid groups in the structure of the enzyme. The results indicated that, compared with the other surfactants, activating CRL with calix[4]-L-proline resulted in hyperactivation at all concentrations (a relative increase of 230%).
Topics: Anions; Biocatalysis; Calixarenes; Candida; Catalytic Domain; Detergents; Enzymes, Immobilized; Hydrogen Bonding; Hydrogen-Ion Concentration; Hydrolysis; Kinetics; Lipase; Magnetic Resonance Spectroscopy; Phenols; Proline; Saccharomycetales; Sodium Dodecyl Sulfate; Static Electricity; Stereoisomerism; Surface-Active Agents
PubMed: 32601811
DOI: 10.1007/s00449-020-02397-3