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Molecules (Basel, Switzerland) May 2024The development of immobilized enzymes with high activity and stability is critical. Metal-organic frameworks (MOFs) have attracted much academic and industrial interest...
The development of immobilized enzymes with high activity and stability is critical. Metal-organic frameworks (MOFs) have attracted much academic and industrial interest in the field of enzyme immobilization due to their unique properties. In this study, the amino-functionalized ionic liquid (NIL)-modified metal-organic framework (UiO-66-NH) was prepared to immobilize lipase (CRL), using dialdehyde starch (DAS) as the cross-linker. The results of the Fourier transform infrared (FT-IR) spectra, X-ray powder diffraction (XRD), and scanning electronic microscopy (SEM) confirmed that the NIL was successfully grafted to UiO-66-NH. The CRL immobilized on NIL-modified UiO-66-NH (UiO-66-NH-NIL-DAS@CRL) exhibited satisfactory activity recovery (79.33%), stability, reusability, and excellent organic solvent tolerance. The research results indicated that ionic liquid-modified UiO-66-NH had practical potential for application in enzyme immobilization.
Topics: Lipase; Ionic Liquids; Enzymes, Immobilized; Metal-Organic Frameworks; Enzyme Stability; Spectroscopy, Fourier Transform Infrared; X-Ray Diffraction; Starch; Saccharomycetales; Phthalic Acids
PubMed: 38792242
DOI: 10.3390/molecules29102381 -
International Journal of Molecular... May 2024The use of lipase immobilized on an octyl-agarose support to obtain the optically pure enantiomers of chiral drugs in reactions carried out in organic solvents is a...
The use of lipase immobilized on an octyl-agarose support to obtain the optically pure enantiomers of chiral drugs in reactions carried out in organic solvents is a great challenge for chemical and pharmaceutical sciences. Therefore, it is extremely important to develop optimal procedures to achieve a high enantioselectivity of the biocatalysts in the organic medium. Our paper describes a new approach to biocatalysis performed in an organic solvent with the use of CALB-octyl-agarose support including the application of a polypropylene reactor, an appropriate buffer for immobilization (Tris base-pH 9, 100 mM), a drying step, and then the storage of immobilized lipases in a climatic chamber or a refrigerator. An immobilized lipase B from (CALB) was used in the kinetic resolution of (,)-flurbiprofen by enantioselective esterification with methanol, reaching a high enantiomeric excess (ee = 89.6 ± 2.0%). As part of the immobilization optimization, the influence of different buffers was investigated. The effect of the reactor material and the reaction medium on the lipase activity was also studied. Moreover, the stability of the immobilized lipases: lipase from (CRL) and CALB during storage in various temperature and humidity conditions (climatic chamber and refrigerator) was tested. The application of the immobilized CALB in a polypropylene reactor allowed for receiving over 9-fold higher conversion values compared to the results achieved when conducting the reaction in a glass reactor, as well as approximately 30-fold higher conversion values in comparison with free lipase. The good stability of the CALB-octyl-agarose support was demonstrated. After 7 days of storage in a climatic chamber or refrigerator (with protection from humidity) approximately 60% higher conversion values were obtained compared to the results observed for the immobilized form that had not been stored. The new approach involving the application of the CALB-octyl-agarose support for reactions performed in organic solvents indicates a significant role of the polymer reactor material being used in achieving high catalytic activity.
Topics: Lipase; Enzymes, Immobilized; Biocatalysis; Fungal Proteins; Sepharose; Propionates; Stereoisomerism; Kinetics; Esterification; Temperature; Enzyme Stability; Candida; Solvents; Saccharomycetales
PubMed: 38791124
DOI: 10.3390/ijms25105084 -
Journal of Biotechnology May 2024Enzyme immobilization in membrane bioreactors has been considered as a practical approach to enhance the stability, reusability, and efficiency of enzymes. In this...
Enzyme immobilization in membrane bioreactors has been considered as a practical approach to enhance the stability, reusability, and efficiency of enzymes. In this particular study, a new type of hybrid membrane reactor was created through the phase inversion method, utilizing hybrid of graphene oxide nanosheets (GON) and polyether sulfone (PES) in order to covalently immobilize the Candida rugosa lipase (CRL). The surface of hybrid membrane was initially modified by (3-Aminopropyl) triethoxysilane (APTES), before the use of glutaraldehyde (GLU), as a linker, through the imine bonds. The resulted enzymatic hybrid membrane reactors (EHMRs) were then thoroughly analyzed by using field-emission scanning electron microscopy (FE-SEM), contact angle goniometry, surface free energy analysis, X-ray diffraction (XRD), Fourier transform infrared (FT-IR) spectroscopy, attenuated total reflection (ATR), and energy-dispersive X-ray (EDX) spectroscopy. The study also looked into the impact of factors such as initial CRL concentration, storage conditions, and immobilization time on the EHMR's performance and activity, which were subsequently optimized. The results demonstrated that the CRLs covalently immobilized on the EHMRs displayed enhanced pH and thermal stability compared to those physically immobilized or free. These covalently immobilized CRLs could maintain over 60% of their activity even after 6 reaction cycles spanning 50 days. EHMRs are valuable biocatalysts in developing various industrial, environmental, and analytical processes.
Topics: Enzymes, Immobilized; Lipase; Enzyme Stability; Bioreactors; Membranes, Artificial; Graphite; Saccharomycetales; Glutaral; Spectroscopy, Fourier Transform Infrared; Fungal Proteins; Temperature; X-Ray Diffraction
PubMed: 38548020
DOI: 10.1016/j.jbiotec.2024.03.012 -
Bioresource Technology May 2024This study established a Candida rugosa lipase (CRL) system to catalyze triolein and ethyl ferulate interesterification. The products were identified, and the binding...
This study established a Candida rugosa lipase (CRL) system to catalyze triolein and ethyl ferulate interesterification. The products were identified, and the binding mode between the substrates and CRL was predicted through molecular docking. Three methods for preparing CRL-AuNPs were proposed and characterized. It was found that the addition of 40 mL of 15 nm gold nanoparticles increased the CRL activity from 3.05 U/mg to 4.75 U/mg, but the hybridization efficiency was only 32.7 %. By using 4 mL of 0.1 mg/mL chloroauric acid, the hybridization efficiency was improved to 50.7 %, but the enzyme activity was sharply decreased. However, when the molar ratio of Mb to HAuCl was 0.2, the hybridization efficiency increased to 71.8 %, and the CRL activity was also enhanced to 5.98 U/mg. Under optimal conditions, the enzyme activity of CRL-AuNPs③ was maintained at 95 % after 6 repetitions and 85.6 % after 30 days at room temperature.
Topics: Lipase; Gold; Enzymes, Immobilized; Triolein; Molecular Docking Simulation; Candida; Metal Nanoparticles; Enzyme Stability; Caffeic Acids; Saccharomycetales
PubMed: 38493938
DOI: 10.1016/j.biortech.2024.130599 -
Veterinary Clinical Pathology Jun 2024A 6-year-old 21.5 kg castrated male Siberian Husky was presented for acute onset of lethargy, vomiting, hemorrhagic diarrhea, and inappetence. Physical examination...
A 6-year-old 21.5 kg castrated male Siberian Husky was presented for acute onset of lethargy, vomiting, hemorrhagic diarrhea, and inappetence. Physical examination revealed marked discomfort upon abdominal palpation and 5%-7% dehydration. The CBC and biochemical profile revealed changes consistent with mild to moderate inflammation, dehydration, and gastrointestinal (GI) disease. Despite aggressive gastrointestinal support, anorexia persisted, and an upper GI endoscopy was performed in conjunction with esophagostomy tube placement. Endoscopy revealed abnormal gastric mucosa characterized by moderately well-demarcated areas of blue-black discoloration. Impression smears of a gastric biopsy revealed abundant extracellular yeasts with morphology most consistent with Candida spp. and frequent extracellular cocci. Similar yeast and bacteria, in lower numbers, were observed on cytologic analysis of a direct smear of the rectal mucosa. A rectal swab submitted for fungal culture yielded pure growth of fungal yeasts identified as Diutina (formerly Candida) rugosa by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry. The dog's clinical signs improved with fluconazole, and he was discharged. Follow-up fungal culture of a rectal swab showed no growth of D. rugosa. To the authors' knowledge, this is the first case report that describes the clinical, hematologic, cytologic, and gross findings of enteric colonization by D. rugosa in a dog.
Topics: Animals; Male; Dogs; Dog Diseases; Candida; Candidiasis; Antifungal Agents; Fluconazole
PubMed: 38418373
DOI: 10.1111/vcp.13335 -
Microbial Biotechnology Feb 2024The yeast Komagataella phaffii (Pichia pastoris) is currently considered a versatile and highly efficient host for recombinant protein production (RPP). Interestingly,...
The yeast Komagataella phaffii (Pichia pastoris) is currently considered a versatile and highly efficient host for recombinant protein production (RPP). Interestingly, the regulated application of specific stress factors as part of bioprocess engineering strategies has proven potential for increasing the production of recombinant products. This study aims to evaluate the impact of controlled oxygen-limiting conditions on the performance of K. phaffii bioprocesses for RPP in combination with the specific growth rate (μ) in fed-batch cultivations. In this work, Candida rugosa lipase 1 (Crl1) production, regulated by the constitutive GAP promoter, growing at different nominal μ (0.030, 0.065, 0.100 and 0.120 h ) under both normoxic and hypoxic conditions in carbon-limiting fed-batch cultures is analysed. Hypoxic fermentations were controlled at a target respiratory quotient (RQ) of 1.4, with excellent performance, using an innovative automated control based on the stirring rate as the manipulated variable developed during this study. The results conclude that oxygen limitation positively affects bioprocess efficiency under all growing conditions compared. The shift from respiratory to respiro-fermentative metabolism increases bioprocess productivity by up to twofold for the specific growth rates evaluated. Moreover, the specific product generation rate (q ) increases linearly with μ, regardless of oxygen availability. Furthermore, this hypoxic boosting effect was also observed in the production of Candida antarctica lipase B (CalB) and pro-Rhizopus oryzae lipase (proRol), thus proving the synergic effect of kinetic and physiological stress control. Finally, the Crl1 production scale-up was conducted successfully, confirming the strategy's scalability and the robustness of the results obtained at the bench-scale level.
Topics: Pichia; Recombinant Proteins; Lipase; Oxygen; Saccharomycetales
PubMed: 38376073
DOI: 10.1111/1751-7915.14411 -
Mikrochimica Acta Nov 2023A sensitive and accurate chemiluminescence (CL) method was developed for one-step determination of diphenyl ether herbicides at trace level with nitrofen...
A sensitive and accurate chemiluminescence (CL) method was developed for one-step determination of diphenyl ether herbicides at trace level with nitrofen (2,4-dichlorophenyl-p-nitrophenyl ether) as a model analyte. Candida rugosa lipase (CRL) was immobilized on a nanocarrier of amine-linked covalent organic framework (named as COF-300-AR) through a self-assembly strategy. The formed nanocomposite of COF-300-AR@CRL owns dual enzymatic catalytic activities. It can directly catalyze luminol-dissolved oxygen reaction to produce an intense CL emission by virtue of oxidase mimic activity of COF-300-AR but also effectively decompose nitrofen to release phenolic compounds by the immobilized CRL. The released phenolic compounds own strong reducing capacity and in turn decrease the CL signal sharply. Under the optimal conditions, the decreased CL intensity presents a good linear response to nitrofen concentration in the 0.02-50.0 μM range. The limit of detection (LOD, 3s/S) is 11 nM and the precision is 2.0% for replicate measurements of 50.0 nM nitrofen solution (n = 11). This method has the advantages of rapid analytical efficiency, good selectivity, satisfactory stability, and recyclability. Recovery experiments were conducted on spiked vegetable and fruit samples with the recoveries falling in the range 90.0-107.0%.
Topics: Vegetables; Fruit; Phenyl Ethers; Phenols; Herbicides; Lipase
PubMed: 38032482
DOI: 10.1007/s00604-023-06077-3 -
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 -
International Journal of Biological... Dec 2023Lipase adsorption on solid supports can be mediated by a precise balance of electrostatic and hydrophobic interactions. A suitable fine-tuning could allow the...
Lipase adsorption on solid supports can be mediated by a precise balance of electrostatic and hydrophobic interactions. A suitable fine-tuning could allow the immobilized enzyme to display high catalytic activity. The objective of this work was to investigate how pH and ionic strength fluctuations affected protein-support interactions during immobilization via physical adsorption of a Candida rugosa lipase (CRL) on MgFeO. The highest amount of immobilized protein (IP) was measured at pH 4, and an ionic strength of 90 mM. However, these immobilization conditions did not register the highest hydrolytic activity (HA) in the biocatalyst (CRLa@MgFeO), finding the best values also at acidic pH but with a slight shift towards higher values of ionic strength around 110 mM. These findings were confirmed when the adsorption isotherms were examined under different immobilization conditions so that the maximum measurements of IP did not coincide with that of HA. Furthermore, when the recovered activity was examined, a strong interfacial hyperactivation of the lipase was detected towards acidic pH and highly charged surrounding environments. Spectroscopic studies, as well as in silico molecular docking analyses, revealed a considerable involvement of surface hydrophobic protein-carrier interactions, with aromatic aminoacids, especially phenylalanine residues, playing an important role. In light of these findings, this study significantly contributes to the body of knowledge and a better understanding of the factors that influence the lipase immobilization process on magnetic inorganic oxide nanoparticle surfaces.
Topics: Lipase; Molecular Docking Simulation; Candida; Enzymes, Immobilized; Nanoparticles; Enzyme Stability
PubMed: 37652323
DOI: 10.1016/j.ijbiomac.2023.126615 -
Bioengineering (Basel, Switzerland) Aug 2023This work demonstrated the feasibility of an industrial-scale aerated static pile composting system for treating one of the common biowastes-soybean curd residue. The...
This work demonstrated the feasibility of an industrial-scale aerated static pile composting system for treating one of the common biowastes-soybean curd residue. The mixing ratios of the feedstock were optimized to achieve a carbon-nitrogen ratio and a moisture level in the ranges of 25-35 and 60-70%, respectively. This open-air composting system required 6-7 months to obtain a mature compost. Solvita and seed germination tests further confirmed the maturity of the compost, with 25% compost extract concentration yielding the best germination index in the absence of phytotoxicity. The bacterial and fungal compositions of the compost piles were further examined with metagenomic analysis. spp., spp., and spp. were among the unique bacteria found, and , , and were among the unique fungi found in the compost piles, suggesting the presence of good microorganisms for degrading the organic biowastes.
PubMed: 37627823
DOI: 10.3390/bioengineering10080938