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1.
Int J Biol Macromol ; 274(Pt 1): 133236, 2024 Aug.
Article in English | MEDLINE | ID: mdl-38897511

ABSTRACT

Nucleating agents, especially those with small particle sizes, are preferred to boost the nucleation density and crystallinity of poly(lactic acid) (PLA) due to its weak crystallization capability. Organophilicly modified nanofillers hardly alter the nucleation and crystallinity of non-isothermally crystallized PLA. Herein, nano-silica adsorbed trace D-sorbitol (m-SiO2) as a heterogeneous nucleating agent was melt-mixed with poly(L-lactic acid) (PLLA), and the isothermal and non-isothermal crystallization behavior, as well as crystallization kinetics, were investigated. Transmission electron microscopy (TEM) revealed that m-SiO2 was uniformly dispersed in the PLA matrix as 100-300 nm clusters. Differential scanning calorimetry (DSC) and polarized optical microscopy (POM) showed that the nucleation rate and density of the non-isothermally crystallized PLLA/m-SiO2 composites were significantly improved. Despite the fact that m-SiO2 does not raise the overall non-isothermal crystallization rate, the crystallization temperature and crystallinity of the PLLA/3%m-SiO2 composite increased from 97.2 °C and 6.8 % for neat PLLA to 108.2 °C and 48.6 % (10 °C/min cooling rate), respectively. The Avrami exponent n of isothermal crystallization remains unchanged, while the crystallization rate increases dramatically. Both isothermal and non-isothermal crystallization have increased activation energies. The heat deflection temperature increased from 59 °C of neat PLLA to 152 °C with a 50 % increase in impact strength.


Subject(s)
Crystallization , Polyesters , Silicon Dioxide , Sorbitol , Polyesters/chemistry , Silicon Dioxide/chemistry , Sorbitol/chemistry , Nanoparticles/chemistry , Calorimetry, Differential Scanning , Kinetics , Temperature
2.
Int J Biol Macromol ; 265(Pt 1): 130902, 2024 Apr.
Article in English | MEDLINE | ID: mdl-38492697

ABSTRACT

The preparation of bio-based poly(lactic acid) (PLA) foams with high mechanical properties and heat resistance is of great significance for environmental protection and green sustainable development. In this paper, D-sorbitol (DS) containing six hydroxyl groups was introduced into poly(l-lactide) (PLLA)/poly(d-lactide) (PDLA) blends for first time to promote the formation of stereocomplex (SC) crystals, which could improve the foaming behavior and enhance mechanical properties and heat resistance of PLA foams. The results showed that DS could improve the formation efficiency and crystallinity of SC crystals by enhancing the hydrogen bonding between the enantiomeric molecular chains. Furthermore, the compression modulus and interactions Vicat softening temperature of the PLLA/PDLA/DS blend foam increased about 854% and 16% compared to the pure PLLA foam, respectively. Besides, when the annealing process was introduced, the compression and heat resistance of the PLA foams increased further. This study provided a feasible strategy for the preparation of bio-based and biodegradable PLA foams with outstanding compressive and heat resistance properties.


Subject(s)
Hot Temperature , Polymers , Polymers/chemistry , Crystallization , Polyesters/chemistry
3.
Adv Mater ; 36(23): e2313157, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38421078

ABSTRACT

Electrophysiology, exploring vital electrical phenomena in living organisms, anticipates broader integration into daily life through wearable devices and epidermal electrodes. However, addressing the challenges of the electrode durability and motion artifacts is essential to enable continuous and long-term biopotential signal monitoring, presenting a hurdle for its seamless implementation in daily life. To address these challenges, an ultrathin polymeric conductive adhesive, poly(3,4-ethylenedioxythiophene): poly(styrenesulfonate)/polyvinyl alcohol/d-sorbitol (PPd) electrode with enhanced adhesion, stretchability, and skin conformability, is presented. The skin conformability and stability of electrodes is designed by theoretical criteria obtained by mechanical analysis. Thus, impedance stability is obtained over 1-week of daily life, and the PPd electrode addresses the challenges related to durability during prolonged usage. Proving stability in electromyography (EMG) signals during high-intensity exercise, the wireless PPd measurement system exhibits high signal-to-noise ratio (SNR) signals even in situations involving significant and repetitive skin deformation. Throughout continuous 1 week-long electrocardiogram (ECG) monitoring in daily life, the system consistently preserves signal quality, underscoring the heightened durability and applicability of the PPd measurement system.


Subject(s)
Adhesives , Electric Conductivity , Electrodes , Adhesives/chemistry , Humans , Wearable Electronic Devices , Polystyrenes/chemistry , Polymers/chemistry , Epidermis/physiology , Electromyography/methods , Polyvinyl Alcohol/chemistry , Electrocardiography , Signal-To-Noise Ratio , Bridged Bicyclo Compounds, Heterocyclic/chemistry , Monitoring, Physiologic/instrumentation , Monitoring, Physiologic/methods , Thiophenes/chemistry
4.
BMC Psychiatry ; 24(1): 81, 2024 Jan 30.
Article in English | MEDLINE | ID: mdl-38291403

ABSTRACT

BACKGROUND: Asenapine has unique orally-related side effects, such as a bitter taste induced by sublingual administration, which often results in discontinuation of the medication. While the FDA has approved black-cherry-flavored asenapine, several countries have prescribed only unflavored versions. Specifically, Asians commonly report experiencing the bitterness of asenapine because they are more sensitive to bitter tastes than other ethnic groups. In this study, with the aim of improving adherence by reducing the bitterness of asenapine, we investigated the effects of D-sorbitol, which reduced the bitterness parameters of taste sensors in our previous basic study on the bitterness and continuity of asenapine among patients with schizophrenia. METHODS: Twenty adult patients with schizophrenia were included in this single-blind, placebo-controlled, crossover trial. Participants rinsed their mouths with single-administration of D-sorbitol or a placebo prior to each administration of asenapine. We then conducted the questionnaires and assessed changes in the bitterness of asenapine (primary end point) and willingness to continue its use (secondary end point). RESULTS: D-sorbitol significantly improved the bitterness of asenapine (p = 0.038). Although it did not significantly increase the willingness to continue asenapine (p = 0.180), it did show improvement over the placebo in enhancing willingness to continue, especially in patients who were not accustomed to its taste. CONCLUSION: Our findings indicate that single-administration of D-sorbitol significantly reduces the bitterness of asenapine. In countries where flavored asenapine is not available, this finding could benefit patients who were not accustomed to its bitter taste. TRIAL REGISTRATION: This study was registered in the Japan Registry of Clinical Trials (jRCTs041210019) on May 14, 2021.


Subject(s)
Antipsychotic Agents , Dibenzocycloheptenes , Adult , Humans , Antipsychotic Agents/adverse effects , Taste , Single-Blind Method , Cross-Over Studies , Heterocyclic Compounds, 4 or More Rings/pharmacology , Heterocyclic Compounds, 4 or More Rings/therapeutic use , Treatment Outcome
5.
Foods ; 12(9)2023 May 02.
Article in English | MEDLINE | ID: mdl-37174415

ABSTRACT

Sustainability, low toxicity, and high solute potential are the fundamental reasons for focusing green chemistry on natural deep eutectic solvents (NADES). The application of NADES ranges from organic chemistry to the agricultural sector and the food industry. In the food industry, the desired food quality can be achieved by the extraction of small molecules, macromolecules, and even heavy metals. The compound yield in Maillard-type model reactions can also be increased using NADES. To extend the so-called "kitchen-type chemistry" field, an inert, food-grade NADES system based on sucrose/D-sorbitol was developed, characterized, and examined for its ability as a reaction medium by evaluating its temperature and pH stability. Reaction boundary conditions were determined at 100 °C for three hours with a pH range of 3.7-9.0. As proof of principle, two Maillard-type model reactions were implemented to generate the taste-modulating compounds N2-(1-carboxyethyl)guanosine 5'-monophosphate) (161.8 µmol/mmol) and N2-(furfuryl thiomethyl)guanosine 5'-monophosphate (95.7 µmol/g). Since the yields of both compounds are higher than their respective taste-modulating thresholds, the newly developed NADES is well-suited for these types of "kitchen-type chemistry" and, therefore, a potential solvent candidate for a wide range of applications in the food industry.

6.
BMC Psychiatry ; 23(1): 159, 2023 03 14.
Article in English | MEDLINE | ID: mdl-36918838

ABSTRACT

BACKGROUND: Antipsychotics are essential in the acute treatment of and maintenance therapy for schizophrenia, but medication adherence and long-term treatment continuity are needed to maximize their effectiveness. Each antipsychotic has various side effects, which may affect adherence. Some patients with schizophrenia are reluctant to take asenapine because of its unique oral-related side effects, such as the bitter taste caused by sublingual administration. Our previous basic research found that D-sorbitol lowered the bitterness parameters of the taste sensors. However, whether D-sorbitol has the same effect in patients remains unclear. Therefore, using a D-sorbitol solution, we aim to evaluate changes in the bitterness of asenapine among patients with schizophrenia. METHODS: In this single-blind, placebo-controlled, crossover trial, we plan to recruit 20 adult patients with schizophrenia spectrum disorder who take sublingual asenapine tablets. The participants will be divided into two groups (n = 10 each). Each group will be given a D-sorbitol or placebo solution on the first day for rinsing before taking the sublingual asenapine tablets. After a 1-day interval, the participants will rinse their mouths again with a different liquid. Questionnaires regarding changes in taste and the willingness to continue asenapine will be conducted before the start of the study and after each rinse. The primary and secondary end points will be a taste evaluation of bitterness, and the willingness to continue asenapine, respectively. Differences in questionnaire scores between the D-sorbitol and placebo solutions will be calculated and analyzed using a McNemar test. DISCUSSION: This study aims to determine the efficacy of D-sorbitol in masking the bitter taste of asenapine. To our knowledge, it is the first intervention study using D-sorbitol for bitter taste of asenapine in patients with schizophrenia. Evidence of the efficacy of D-sorbitol could result in D-sorbitol pretreatment being an easy and inexpensive means of improving adherence to asenapine. TRIAL REGISTRATION: This study was registered in the Japan Registry of Clinical Trials jRCTs041210019, on May 14, 2021. Ethics approval was obtained from the Nagoya University Clinical Research Review Board.


Subject(s)
Antipsychotic Agents , Taste , Adult , Humans , Cross-Over Studies , Single-Blind Method , Heterocyclic Compounds, 4 or More Rings/adverse effects , Treatment Outcome
7.
Brain ; 146(7): 2885-2896, 2023 07 03.
Article in English | MEDLINE | ID: mdl-36511878

ABSTRACT

Charcot-Marie-Tooth disease is the most common inherited disorder of the PNS. CMT1A accounts for 40-50% of all cases and is caused by a duplication of the PMP22 gene on chromosome 17, leading to dysmyelination in the PNS. Patient-derived models to study such myelination defects are lacking as the in vitro generation of human myelinating Schwann cells has proved to be particularly challenging. Here, we present an induced pluripotent stem cell-derived organoid culture, containing various cell types of the PNS, including myelinating human Schwann cells, which mimics the human PNS. Single-cell analysis confirmed the PNS-like cellular composition and provides insight into the developmental trajectory. We used this organoid model to study disease signatures of CMT1A, revealing early ultrastructural myelin alterations, including increased myelin periodic line distance and hypermyelination of small axons. Furthermore, we observed the presence of onion-bulb-like formations in a later developmental stage. These hallmarks were not present in the CMT1A-corrected isogenic line or in a CMT2A iPSC line, supporting the notion that these alterations are specific to CMT1A. Downregulation of PMP22 expression using short-hairpin RNAs or a combinatorial drug consisting of baclofen, naltrexone hydrochloride and D-sorbitol was able to ameliorate the myelin defects in CMT1A-organoids. In summary, this self-organizing organoid model can capture biologically meaningful features of the disease and capture the physiological complexity, forms an excellent model for studying demyelinating diseases and supports the therapeutic approach of reducing PMP22 expression.


Subject(s)
Charcot-Marie-Tooth Disease , Induced Pluripotent Stem Cells , Humans , Myelin Sheath/metabolism , Induced Pluripotent Stem Cells/metabolism , Down-Regulation , Myelin Proteins/genetics , Myelin Proteins/metabolism , Charcot-Marie-Tooth Disease/genetics , Charcot-Marie-Tooth Disease/metabolism , Organoids/metabolism , Schwann Cells
8.
J Biomed Mater Res B Appl Biomater ; 111(1): 26-37, 2023 01.
Article in English | MEDLINE | ID: mdl-35809250

ABSTRACT

Vitamin E (VE) is currently an approved antioxidant to improve the oxidation stability of highly crosslinked ultrahigh molecular weight polyethylene (UHMWPE) insert used commercially in total joint arthroplasty. However, the decrease in crosslink density caused by VE reduces wear resistance of UHMWPE, showing an uncoordinated challenge. In this work, we hypothesized that D-sorbitol (DS) as a secondary antioxidant can improve the antioxidant efficacy of VE on chemically crosslinked UHMWPE. The combined effect of VE and DS on oxidation stability of UHMWPE was investigated at a set of controlled hybrid antioxidant content. The hybrid antioxidant strategy showed significantly synergistic enhancement on the oxidation stability of chemically crosslinked UHMWPE compared with the single VE strategy. More strikingly, the crosslink density of the blends with hybrid antioxidants stayed at a high level since DS is not sensitive to crosslinking. The relationships between oxidation stability, mechanical properties, crosslink density, and crystallinity were investigated, by which the clinically relevant overall performance of UHMWPE was optimized. This work provides a leading-edge design mean for the development of joint bearings.


Subject(s)
Antioxidants , Polyethylenes , Antioxidants/chemistry , Molecular Weight , Materials Testing , Polyethylenes/chemistry , Vitamin E/chemistry
9.
Yakugaku Zasshi ; 142(4): 401-411, 2022.
Article in Japanese | MEDLINE | ID: mdl-35370196

ABSTRACT

Ophthalmic viscosurgical devices (OVDs) are mainly divided into two general categories: cohesive and dispersive. Dispersive OVDs such as the 3% hyaluronic acid and 4% chondroitin sulfate (HA/CS) combination have excellent adhesion to ocular tissues and protect the corneal endothelium to a greater extent than cohesive OVDs. Herein, we summarize our recent findings regarding one of the properties of the HA/CS combination related to clinical performance. (i) The room temperature stability of OVDs and needle clogging by OVDs remain clinical issues. We demonstrated that adding d-sorbitol to the HA/CS combination preserved its viscosity, which was equivalent after 2 year-storage at room temperature to the viscosity of HA/CS combination stored under refrigeration for 2 years without d-sorbitol. Besides, the HA/CS combination with d-sorbitol could be used repeatedly without cleaning or replacing the needle, suggesting that the addition of d-sorbitol prevents drying and solidification of the OVD on the needle. (ii) Although it can be inferred from numerous studies that the tissue adhesion of OVDs influences their retention by the eye, little is known about the physical properties of OVDs that contribute to intraocular retention. To address this issue, we compared two types of adhesive forces, detachment force and repulsive force, for each OVD. Compared with other dispersive OVDs, the HA/CS combination showed higher values for both adhesive forces. These results suggest that adhesive forces may be used as an index of dispersive OVD retention in the eye.


Subject(s)
Chondroitin Sulfates , Phacoemulsification , Adhesives , Hyaluronic Acid , Viscosity
10.
Synth Syst Biotechnol ; 7(2): 730-737, 2022 Jun.
Article in English | MEDLINE | ID: mdl-35356389

ABSTRACT

l-Sorbose is an essential intermediate for the industrial production of vitamin C (l-ascorbic acid). However, the formation of fructose and some unknown by-products significantly reduces the conversion ratio of D-sorbitol to l-sorbose. This study aimed to identify the key D-sorbitol dehydrogenases in Gluconobacter oxydans WSH-003 by gene knockout. Then, a total of 38 dehydrogenases were knocked out in G. oxydans WSH-003, and 23 dehydrogenase-deficient strains could increase l-sorbose production. G. oxydans-30, wherein a pyrroloquinoline quinone-dependent glucose dehydrogenase was deleted, showed a significant reduction of a by-product with the extension of fermentation time. In addition, the highest conversion ratio of 99.60% was achieved in G. oxydans MD-16, in which 16 different types of dehydrogenases were inactivated consecutively. Finally, the gene vhb encoding hemoglobin was introduced into the strain. The titer of l-sorbose was 298.61 g/L in a 5-L bioreactor. The results showed that the systematic engineering of dehydrogenase could significantly enhance the production of l-sorbose.

11.
J Biotechnol ; 346: 1-10, 2022 Feb 20.
Article in English | MEDLINE | ID: mdl-35038459

ABSTRACT

Erythropoietin (EPO) is a glycoprotein hormone that has been used to treat anemia in patients with chronic kidney disease and in cancer patients who are receiving chemotherapy. Here, we investigated the accessibility of the glutamine (Gln, Q) residues of recombinant human erythropoietin (rHuEPO) towards a thermoresistant variant microbial transglutaminase (mTGase), TG16 with the aim of developing novel rHuEPO conjugates that may potentially enhance its biological efficacy. As a model bioconjugation, we studied the reactivity of rHuEPO towards TG16 with a low molar mass amine group containing substrate, monodansyl cadaverine (MDC). The reactions were carried out at a Tm of 54.3 °C, the transition temperature of rHuEPO. Characterization by SDS-PAGE and mass spectrometry confirmed the conjugates formation. Then, we examined the conjugation of rHuEPO with a biodegradable and biocompatible polyester, poly(D-sorbitol adipate) (PDSA). To achieve this, PDSA was enzymatically synthesized using lipase B from Candida antartica (CAL-B), chemically modified with side chains having free primary amine (NH2) groups that can be acyl acceptor substrate of TG16, thoroughly characterized by 1H NMR spectroscopy, and then applied for the TG16-mediated conjugation reaction with rHuEPO. rHuEPO conjugates generated by this approach were identified by SDS-PAGE proving that the amine-grafted PDSA is accepted as a substrate for TG16. The successful conjugation was further verified by the detection of high molar mass fluorescent bands after labelling of amine-grafted PDSA with rhodamine B-isothiocyanate. Overall, this enzymatic procedure is considered as an effective approach to prepare biodegradable rHuEPO-polymer conjugates even in the presence of N- and O-glycans.


Subject(s)
Anemia , Erythropoietin , Anemia/drug therapy , Humans , Polyesters , Recombinant Proteins/therapeutic use , Transglutaminases
12.
Biochem Biophys Res Commun ; 586: 81-86, 2022 01 01.
Article in English | MEDLINE | ID: mdl-34837836

ABSTRACT

1-Deoxy-D-sorbitol, the 1-deoxy analogue of D-sorbitol, has been detected in human urine as well as in natural herbs and spices. Although there are sporadic reports about 1-deoxy-D-sorbitol dehydrogenase, the complete catabolic pathway of 1-deoxy-D-sorbitol remains unsolved. Informed by the promiscuous activities of fructose-6-phosphate aldolase (FSA) which is involved in the sorbitol (glucitol) utilization (gut) operon and guided by the large scale bioinformatics analysis, we predicted and then experimentally verified the gut operon encoded by Bacillus licheniformis ATCC14580 is responsible for the catabolism of both D-sorbitol and 1-deoxy-D-sorbitol by in vitro activity assays of pathway enzymes, in vivo growth phenotypes, and transcriptomic studies. Moreover, the phylogenetic distribution analysis suggests that the D-sorbitol and 1-deoxy-D-sorbitol catabolic gene cluster is mostly conserved in members of Firmicutes phylum.


Subject(s)
Aldehyde-Lyases/metabolism , Bacillus licheniformis/metabolism , Bacterial Proteins/metabolism , Metabolism/genetics , Sorbitol/metabolism , Aldehyde-Lyases/genetics , Bacillus licheniformis/classification , Bacillus licheniformis/genetics , Bacterial Proteins/genetics , Computational Biology/methods , Gene Expression Regulation, Bacterial , Glycerol/chemistry , Glycerol/metabolism , Mannitol/chemistry , Mannitol/metabolism , Operon , Phylogeny , Sorbitol/analogs & derivatives
13.
Acta Biomater ; 134: 302-312, 2021 10 15.
Article in English | MEDLINE | ID: mdl-34311104

ABSTRACT

Oxidative stability of radiation crosslinked ultrahigh molecular weight polyethylene (UHMWPE) artificial joints is significantly improved by vitamin E (VE), but there is a dilemma that VE hinders crosslinking and thus jeopardizes the wear of UHMWPE. In this effort, we proposed an efficient strategy to stabilize UHMWPE under limited antioxidant contents, where VE and D-sorbitol (DS) were used as the primary antioxidant and the secondary antioxidant respectively. For non-irradiated blends with fixed antioxidant contents, oxidative stability accessed by oxidation induction time (OIT) of VE/DS/UHMWPE blends was superior to that of VE/UHMWPE blends, while DS/UHMWPE blends showed no increase in OIT. The cooperation between DS and VE exhibited a synergistic effect on enhancing the oxidative stability of UHMWPE. Interestingly, the irradiated VE/DS/UHMWPE blends showed comparable OIT but a significantly higher crosslink density than the irradiated VE/UHMWPE blends. The crystallinity, melting point, and in vitro biocompatibility of the blends were not affected by VE and DS. The quantitative relationships of mechanical properties, oxidation stability, crystallinity and crosslink density were established to unveil the correlation of these key factors. The overall properties of VE/UHMWPE and VE/DS/UHMWPE blends were compared to elucidate the superiority of the antioxidant compounding strategy. These findings provide a paradigm to break the trade-off between oxidative stability, crosslink density and mechanical properties, which is constructive to develop UHMWPE bearings with upgraded performance for total joint replacements. STATEMENT OF SIGNIFICANCE: VE-stabilized UHMWPE is the most commonly used material in total joint replacements at present. However, oxidation and wear resistance of VE/UHMWPE implants cannot be unified since VE reduces the efficiency of radiation crosslinking. It limits the use of VE. Herein, we proposed a compounding stabilization by the synergy between VE and DS. The antioxidation capability of VE was revived by DS, thus enhancing the oxidation stability of unirradiated UHMWPE. The irradiated VE/DS/UHMWPE exhibited similar oxidation stability but higher crosslink density than irradiated VE/UHMWPE, which is beneficial to combat wear of UHMWPE and to inhibit the occurrence of osteolysis. This synergistic antioxidation strategy endows the UHMWPE joint material with good overall performance, which is of clinical significance.


Subject(s)
Polyethylenes , Vitamin E , Materials Testing , Molecular Weight , Sorbitol , Vitamin E/pharmacology
14.
Molecules ; 26(10)2021 May 18.
Article in English | MEDLINE | ID: mdl-34070087

ABSTRACT

Fused filament fabrication (FFF) is a process used to manufacture oral forms adapted to the needs of patients. Polyethylene oxide (PEO) filaments were produced by hot melt extrusion (HME) to obtain a filament suitable for the production of amiodarone hydrochloride oral forms by FFF 3D printing. In order to produce personalized oral forms adapted to the patient characteristics, filaments used by FFF must be controlled in terms of mass homogeneity along filament. This work highlights the relation between filament mass homogeneity and its diameter. This is why the impact of filler excipients physical properties was studied. It has been showed that the particle's size distribution of the filler can modify the filament diameter variability which has had an impact on the mass of oral forms produced by FFF. Through this work it was shown that D-Sorbitol from Carlo Erba allows to obtain a diameter variability of less than 2% due to its unique particle's size distribution. Using the filament produced by HME and an innovating calibration method based on the filament length, it has been possible to carry out three dosages of 125 mg, 750 mg and 1000 mg by 3D printing with acceptable mass uniformity.


Subject(s)
Physical Phenomena , Precision Medicine , Printing, Three-Dimensional , Sorbitol/chemistry , Calorimetry, Differential Scanning , Particle Size , Stress, Mechanical , Tensile Strength , Thermogravimetry , Time Factors , X-Ray Diffraction
15.
FEMS Microbiol Lett ; 368(2)2021 02 04.
Article in English | MEDLINE | ID: mdl-33399820

ABSTRACT

Sugar alcohols (polyols) have important roles as nutrients, anti-freezing agents and scavengers of free radicals in cold-adapted bacteria, but the characteristics of polyol dehydrogenases in cold-adapted bacteria remain largely unknown. In this study, based on the observation that a cold-adapted bacterium Pseudomonas mandelii JR-1 predominantly utilized d-sorbitol as its carbon source, among the four polyols examined (d-galactitol, d-mannitol, d-sorbitol and d-xylitol), we cloned and characterized a sorbitol dehydrogenase (SDH, EC 1.1.1.14) belonging to the short-chain dehydrogenase/reductase family from this bacterium (the SDH hereafter referred to as PmSDH). PmSDH contained Asn111, Ser140, Tyr153 and Lys157 as catalytic active site residues and existed as an ∼67-kDa dimer in size-exclusion chromatography. PmSDH converted d-sorbitol to d-fructose using nicotinamide adenine dinucleotide (NAD+) as a cofactor and, vice versa, d-fructose to d-sorbitol using nicotinamide adenine dinucleotide reduced (NADH) as a cofactor. PmSDH maintained its conformational flexibility, secondary and tertiary structures, and thermal stability at 4-25°C. These results indicate that PmSDH, which has a flexible structure and a high catalytic activity at colder temperatures, is well suited to sorbitol utilization in the cold-adapted bacterium P. mandelii JR-1.


Subject(s)
Adaptation, Physiological/genetics , Cold Temperature , L-Iditol 2-Dehydrogenase/genetics , L-Iditol 2-Dehydrogenase/metabolism , Pseudomonas/enzymology , Pseudomonas/genetics , Cloning, Molecular , NAD/metabolism , Protein Structure, Secondary , Protein Structure, Tertiary
16.
J Biosci Bioeng ; 131(5): 518-524, 2021 May.
Article in English | MEDLINE | ID: mdl-33487552

ABSTRACT

The miglitol intermediate, 6-(N-hydroxyethyl)-amino-6-deoxy-α-l-sorbofuranose (6NSL), is catalyzed from N-2-hydroxyethyl glucamine (NHEG) by resting cells of Gluconobacter oxydans. One of the key factors limiting 6NSL production was the availability of oxygen during both cell cultivation and biotransformation of NHEG to 6NSL. Based on G. oxydans/pBBR1-sldAB-pqqABCDE-tldD (G. oxydans/AB-PQQ), the Vitreoscilla hemoglobin (VHb) was heterologously expressed in G. oxydans to enhance oxygen transfer efficiency and improve 6NSL production. The recombinant G. oxydans/AB-PQQ-VHb displayed higher biomass and NHEG oxidation activity than the control stain. The transcription levels of respiratory chain-related enzyme genes in G. oxydans/AB-PQQ-VHb exhibited up-regulation, indicating that the presence of VHb promoted the respiration. The dissolved oxygen (DO) concentration for cell cultivation was optimized in a 5-L stirred bioreactor. At a DO concentration of 20%, the maximum volumetric oxidation activity of NHEG of G. oxydans/AB-PQQ-VHb in the stirred bioreactor reached 168.3 ± 3.2 U/L. Furthermore, the biotransformation of NHEG to 6NSL using G. oxydans/AB-PQQ-VHb was carried out under different oxygen tensions to investigate the effect of oxygen on 6NSL production. Finally, up to 87.5 ± 5.9 g/L 6NSL was accumulated in the reaction mixture within 16 h when the DO was controlled at 30%.


Subject(s)
Bacterial Proteins/genetics , Furans/metabolism , Gluconobacter oxydans/enzymology , L-Iditol 2-Dehydrogenase/genetics , L-Iditol 2-Dehydrogenase/metabolism , Oxygen/metabolism , Protein Engineering , Truncated Hemoglobins/genetics , Bioreactors , Fermentation , Furans/chemistry , Gene Expression , Oxidation-Reduction
17.
Enzyme Microb Technol ; 141: 109670, 2020 Nov.
Article in English | MEDLINE | ID: mdl-33051020

ABSTRACT

6-(N-hydroxyethyl)-amino-6-deoxy-l-sorbofuranose (6NSL), a key precursor in the synthesis of miglitol, is produced from N-2-hydroxyethyl-glucamine (NHEG) by the regioselective oxidation of Gluconobacter oxydans. The limitation of PQQ biosynthesis became a bottleneck for improvement of PQQ-dependent D-sorbitol dehydrogenase (mSLDH) activity. Five expression plasmids were constructed for the co-expression of the pqqABCDE gene cluster and the tldD gene on the basis of pBBR1-gHp0169-sldAB in G. oxydans to increase the biosynthesis of PQQ. The G. oxydans/pGA004, in which pqqABCDE and tldD were expressed as a cluster under the control of gHp0169 promoter, showed the optimal performance. The intracellular PQQ concentration and specific activity of mSLDH in cells increased by 79.3 % and 53.7 %, respectively, compared to that in G. oxydans/pBBR-sldAB. Then, the repeated batch biotransformation of NHEG to 6NSL by G. oxydans/pGA004 was carried out. Up to 75.0 ±â€¯3.0 g/L of 6NSL production with 94.5 ±â€¯3.6 % of average conversion rate of NHEG to 6NSL was achieved after four cycles of run. These results indicated that G. oxydans/pGA004 with high productivity had great potential for 6NSL production in industrial bioprocess.


Subject(s)
Gluconobacter oxydans/metabolism , L-Iditol 2-Dehydrogenase/metabolism , PQQ Cofactor/biosynthesis , Sorbose/analogs & derivatives , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Bioreactors , Biotransformation , Gene Expression , Gluconobacter oxydans/genetics , Gluconobacter oxydans/growth & development , L-Iditol 2-Dehydrogenase/genetics , Multigene Family , Nitrosamines/metabolism , PQQ Cofactor/genetics , PQQ Cofactor/metabolism , Promoter Regions, Genetic , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Sorbose/biosynthesis
18.
ACS Appl Mater Interfaces ; 12(42): 47677-47683, 2020 Oct 21.
Article in English | MEDLINE | ID: mdl-33023290

ABSTRACT

For hole-conductor-free, fully printable mesoscopic perovskite solar cells (MPSCs), it is difficult to achieve free and efficient diffusion of perovskite precursors in micron-scale porous structures. Thus, the wettability of the perovskite precursor is one of the most crucial factors that determine the performance of MPSCs. Here, d-sorbitol hexaacetate (DSHA) is introduced as an additive for fabricating hole-conductor-free, fully printable MPSCs based on methylammonium lead iodide (MAPbI3). The fabricated MPSCs exhibited an efficiency of 14.33%. Moreover, the influence of DSHA on the optical properties, morphology, and filling of perovskite in the MPSCs has been systematically investigated. The results revealed that DSHA effectively optimized the morphology, improved the pore-filling, and passivated the defects of perovskite films. Remarkably, the unencapsulated MPSCs retained 93% of their original power-conversion efficiency (PCE) after 45 days of storage in air with humidity of 50 ± 5%.

19.
J Biotechnol ; 300: 55-62, 2019 Jul 20.
Article in English | MEDLINE | ID: mdl-31100333

ABSTRACT

6-(N-hydroxyethyl) amino-6-deoxy-l-sorbofuranose (6NSL) is the direct precursor of miglitol for diabetes therapy. The regio- and stereo-selective dehydrogenation offered by the membrane-bound d-sorbitol dehydrogenase (mSLDH) from Gluconobacter oxydans provides an elegant enzymatic method for 6NSL production. In this study, two subunits sldA and sldB of mSLDH were introduced into G. oxydans ZJB-605, and the specific enzyme activity of mSLDH towards NHEG was enhanced by 2.15-fold. However, the endogenous PQQ level was dramatically reduced in the recombinant strain and became a bottleneck to support the holo-enzyme activity. A combined supplementation of four amino acids (Glu, Ile, Ser, Arg) involved in biosynthesis of PQQ in conventional media effectively increased extracellular accumulation of PQQ by 1.49-fold, which further enhanced mSLDH activity by 1.33-fold. The synergic improvement of mSLDH activity provided in this study supports the superior high dehydrogenate activity towards substrate N-2-hydroxyethyl-glucamine, 184.28 g·L-1 of 6NSL was produced after a repeated bioconversion process catalyzed by the resting cells of G. oxydans/pBB-sldAB, all of which presenting a great potential of their industrial application in 6NSL biosynthesis.


Subject(s)
Bacterial Proteins/metabolism , Gluconobacter oxydans/metabolism , L-Iditol 2-Dehydrogenase/metabolism , PQQ Cofactor/biosynthesis , Sorbose/analogs & derivatives , 1-Deoxynojirimycin/analogs & derivatives , 1-Deoxynojirimycin/metabolism , Amino Acids/analysis , Bacterial Proteins/genetics , Bioreactors , Culture Media/chemistry , Fermentation , Gene Expression , Gluconobacter oxydans/enzymology , Gluconobacter oxydans/genetics , Hypoglycemic Agents/metabolism , L-Iditol 2-Dehydrogenase/genetics , PQQ Cofactor/metabolism , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Sorbitol/metabolism , Sorbose/biosynthesis
20.
Food Chem ; 289: 436-442, 2019 Aug 15.
Article in English | MEDLINE | ID: mdl-30955634

ABSTRACT

Citric acid, glycerol, and d-sorbitol are used as important food additives. In this research work, viscosities and refractive indices (the physico-chemical properties) for aqueous solution of citric acid, as well as ternary solutions of (water + d-sorbitol + citric acid) and (water + glycerol + citric acid) were measured in mass fractions of citric acid (0.03-0.21) and at temperatures (T = 293.15, 303.15, 313.15, and 323.15) K and atmospheric pressure. For these solutions, the experimental refractive indices were fitted using a semi-empirical equation which its constant, Kr, was introduced by Koohyar in 2011. This constant can be applied to investigate the power of interactions between solute and solvent molecules in aqueous solutions. Also, the experimental viscosities were fitted by the Jones-Dole and Arrhenius-like equations. Obtained data of this research work can be applied in food industries.


Subject(s)
Citric Acid/chemistry , Glycerol/chemistry , Refractometry , Sorbitol/chemistry , Viscosity , Water/chemistry , Atmospheric Pressure , Food Additives , Solutions , Temperature
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