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Summary Black mulberry extract (BME) is rich in phenolics; however, their health benefits are restricted by their instability and poor absorption in the small intestine. Liposomal BME‐loaded pullulan/pectin nanofibers were developed to enhance the in vitro bioaccessibility of BME. The liposomes with BME (0.8%, w/v), were produced by the thin‐film hydration and ultrasonication method with a size of 76.41 ± 1.23 nm and encapsulated 79.40 ± 0.99%.of the BME. Scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM) images showed that the uniform distribution of liposomes within the defect‐free fiber structure. Liposomal BME loading elevated the mucoadhesiveness of the nanofibers compared to free BME loading. Liposomal BME‐loaded nanofiber demonstrated a nearly two‐fold increase in the bioaccessibility of anthocyanins. The cellular release of all four different anthocyanins by Caco‐2 cells was significantly higher (3.92%–10.50%) in liposomal BME‐loaded nanofiber. Therefore, liposomal nanofibers show great potential as a method for delivering phenolics, specifically anthocyanins. [ABSTRACT FROM AUTHOR]

Pullulan is a microbial exopolysaccharide produced by Aureobasidium spp. with excellent physical and chemical properties, resulting in great application value. In this study, a novel strain RM1603 of Aureobasidium pullulans with high pullulan production of 51.0 ± 1.0 g·L− 1 isolated from rhizosphere soil was subjected to atmospheric and room temperature plasma (ARTP) mutagenesis, followed by selection of mutants to obtain pullulan high-producing strains. Finally, two mutants Mu0816 and Mu1519 were obtained, with polysaccharide productions of 58.7 ± 0.8 and 60.0 ± 0.8 g∙L− 1 after 72-h fermentation, representing 15.1 and 17.6% increases compared with the original strain, respectively. Transcriptome analysis of the two mutants and the original strain revealed that the high expression of α/β-hydrolase (ABHD), α-amylase (AMY1), and sugar porter family MFS transporters (SPF-MFS) in the mutants may be related to the synthesis and secretion of pullulan. These results demonstrated the effectiveness of ARTP mutagenesis in A. pullulans, providing a basis for the investigation of genes related to pullulan synthesis and secretion. [ABSTRACT FROM AUTHOR]

Pullulan is a biopolymer produced by various strains of the fungi Aureobasidium pullulans. It is made up of maltotriose units consisting of α-(1,6) and α-(1,4) glycosidic bonds. Pullulan was discovered by Bauer in 1938, characterized by Bernier in 1958 and named as 'pullulan' by Bender in 1959. Submerged fermentation is widely employed for pullulan production followed by downstream processing for recovery of pullulan. Factors such as nutrients, minerals, pH, temperature, surfactants, light intensity, and melanin intermediate affect its production. Various optimization methods such as Plackett Burman design, response surface methodology, and artificial neural networks have been utilised for enhancing the yield of pullulan. Pullulan yield as high as 62.52 g/L can be achieved using a 5 L fermenter under optimised conditions. It can be characterized by techniques such as nuclear magnetic resonance, Fourier transform infrared spectroscopy, X-ray diffraction, scanning electron microscopy. The biopolymer has remarkable properties which makes it an ideal candidate to be used in various applications such as the biomedical and pharmaceutical sectors, cosmetics, food, and wastewater treatment. Pullulan's applications can be expanded by chemically modifying to obtain grafted pullulan, polymer blends, nanocomposites and hydrogels which have been gaining importance in recent years. Over the years, many patents using pullulan have been granted for different applications worldwide. This review sheds light on all these aspects of pullulan including its future scope. [ABSTRACT FROM AUTHOR]

Objectives: The ascomycotic yeast-like fungus Aureobasidium exhibits the natural ability to synthesize several secondary metabolites, like polymalic acid, pullulan, or polyol lipids, with potential biotechnological applications. Combined with its polyextremotolerance, these properties make Aureobasidium a promising production host candidate. Hence, plenty of genomes of Aureobasidia have been sequenced recently. Here, we provide the annotated draft genome sequence of the polyol lipid-producing strain A. pullulans NRRL 62042. Data description: The genome of A. pullulans NRRL 62042 was sequenced using Illumina NovaSeq 6000. Genome assembly revealed a genome size of 24.2 Mb divided into 39 scaffolds with a GC content of 50.1%. Genome annotation using Genemark v4.68 and GenDBE yielded 9,596 genes. [ABSTRACT FROM AUTHOR]

One major limitation of effective vaccine delivery is its dependency on a robust cold chain infrastructure. While Vesicular stomatitis virus (VSV) has been demonstrated to be an effective viral vaccine vector for diseases including Ebola, its −70 °C storage requirement is a significant limitation for accessing disadvantaged locations and populations. Previous work has shown thermal stabilization of viral vaccines with a combination of pullulan and trehalose (PT) dried films. To improve the thermal stability of VSV, we optimized PT formulation concentrations and components, as well as drying methodology with enhanced vacuum drying. When formulated in PT films, VSV can be stored for 32 weeks at 4 °C with less than 2 log PFU loss, at 25 °C with 2.5 log PFU loss, and at 37 °C with 3.1 log PFU loss. These results demonstrate a significant advancement in VSV thermal stabilization, decreasing the cold chain requirements for VSV vectored vaccines. [ABSTRACT FROM AUTHOR]

Pullulan can not only provide a source of organic carbon but also has excellent properties. However, current research is mostly limited to the physical properties of the high-molecular-weight components of pullulan, and little is known about the application of its low-molecular-weight components. This study was designed to explore the impact of presoaking of radish seeds in a pullulan solution on seed germination and subsequent seedling growth under salt stress conditions. Pullulan soaking was found to enhance the germination rates of radish seeds subjected to salt stress, while also enhancing the aboveground growth of radish seedlings. Pullulan soaking resulted in increases in chlorophyll, soluble protein, and soluble sugar concentrations in the leaves of these seedlings, together with greater peroxidase activity and root activity as well as decreases in Na+ and malondialdehyde concentrations. This provides an important reference for the application of pullulan in plant protection. [ABSTRACT FROM AUTHOR]

Pullulan, a natural polysaccharide with unique biocompatibility and biodegradability, has gained prominence in nanomedicine. Its application in nanoparticle drug delivery systems showcases its potential for precision medicine. Aim of Study: This scientific review aims to comprehensively discuss and summarize recent advancements in pullulan-based polymeric nanoparticles, focusing on their formulation, characterization, evaluation, and efficacy. Methodology: A search on Scopus, PubMed, and Google Scholar, using "Pullulan and Nanoparticle" as keywords, identified relevant articles in recent years. Results: The literature search highlighted a diverse range of studies on the pullulan-based polymeric nanoparticles, including the success of high-selectivity hybrid pullulan-based nanoparticles for efficient boron delivery in colon cancer as the active targeting nanoparticle, the specific and high-efficiency release profile of the development of hyalgan-coated pullulan-based nanoparticles, and the design of multifunctional microneedle patches that incorporated pullulan–collagen-based nanoparticle-loaded antimicrobials to accelerate wound healing. These studies collectively underscore the versatility and transformative potential of pullulan-based polymeric nanoparticles in addressing biomedical challenges. Conclusion: Pullulan-based polymeric nanoparticles are promising candidates for innovative drug delivery systems, with the potential to overcome the limitations associated with traditional delivery methods. [ABSTRACT FROM AUTHOR]

A microwave technique was used to prepare a superabsorbent polymer (SAP) by grafting two hydrophilic monomers onto a polysaccharide substrate. The monomers used were acrylic acid (AA) or acrylamide (AM) and were grafted onto a pullulan (PUL) substrate to form PUL-g-AA (SAP1) and PUL-g-AM (SAP2), respectively. The monomers (AM/AA) were grafted together onto a PUL substrate to form PUL-g-(AM/AA) (SAP3). Grafting parameters such as grafting efficiency with the percentage, the conversion of monomer into polymer, gel content, water retention, water adsorption capacity, and swelling kinetics were determined. Additionally, the effect of environmental pH (2, 4, 7, 9, and 12) and sodium dodecylbenzene sulfonate (SDBS) surfactant was evaluated, where 1, 2, 3, 4, and 5 mM of SDBS was added to form SAP4 to SAP8. The FTIR results show that AM was grafted onto PUL through an aliphatic C-N bond, while AA grafting occurred through a single C-C bond. The grafting efficiency with AM was higher than with AA, as well as showing a superior gel content. Water absorbance capacity and water retention increased with the grafting of AA and AM together for SAP3. The highest absorbent capacity, water retention, gel content, and grafting parameters values were obtained with a 3 mM SDBS content and a pH of 7. The swelling kinetics showed that the increases in the theoretical and experimental swelling equilibriums were 72% and 82%, respectively, for SAP6 compared to the values of these parameters for SAP3. The water absorption capacity of the hydrogel increases upon increasing the pH to 7 and then gradually decreases. XRD demonstrated the improved crystallinity and crystalline size of the hydrogel after grafting polymerization of AM/AA onto PUL, in addition to enhanced thermal stability. On the contrary, FE-SEM demonstrated that SDBS improves the porosity and pore size of the hydrogel surface with SAP6. [ABSTRACT FROM AUTHOR]

Novel biocompatible systems suitable for the controlled release of active ingredients are under the spotlight within the last few years. The present investigation focuses on the preparation of nanoparticles (NPs) based on the amphiphilic copolymer pullulan‐graft‐poly(Z‐L‐lysine) bearing different amounts of lysine (10, 20, and 30 wt%), and on the evaluation of their ensuing viability to encapsulate the hydrophobic nonsteroidal anti‐inflammatory drug indomethacin (INDO). The copolymers are synthesized by ring‐opening polymerization, characterized by NMR and FTIR, and their critical aggregation concentration is determined by fluorescence. The NPs are prepared with and without INDO using different copolymer/solvent ratios and INDO/copolymer ratios. The hydrodynamic diameter and polydispersity of the NP suspensions are monitored by dynamic light scattering for 30 days. Their sizes vary between 208 and 338 nm, and some reach the micrometric range (19–73 μm). INDO‐free NPs are identified as spherical‐shaped by atomic force microscopy. Two formulations are tested in terms of encapsulation efficiency (EE = 43%–89%), and the drug crystallinity (DC = 7%–27%) which is determined by differential scanning calorimetry, indicating a reduction with respect to pristine INDO. The results suggest that the copolymers prepared herewith have potential to be applied as carriers for new drug delivery systems of class II drugs. [ABSTRACT FROM AUTHOR]

The objective of this study was to optimize the fermentation parameters by using response surface methodology for increasing pullulan production of Aureobasidium pullulans AZ-6 in synthetic fermentation medium. The optimum conditions were determined as follows: the initial sucrose concentration (X1): 100 g/L; the initial peptone concentration (X2): 11.31 g/L; the initial pH (X3): 6.48; and the temperature (X4): 24.2 °C. In the optimum conditions, the maximum exopolysaccharide (EPS) concentration (Y1), the maximum pullulan concentration (Y2), the specific growth rate (Y3), the maximum specific pullulan formation rate (Y4), and the pullulan yield (Y5) were expected to be obtained as 37.078 and 35.372 g/L, 0.062 h−1, 0.021 [g pullulan/(g mo.h)], and 53.681%, respectively. The observed values of Y1, Y2, Y3, Y4, and Y5 were 36.950 g/L, 35.470 g/L, 0.064 h−1, 0.036 [g pullulan/(g mo.sa)], and 54.480%, respectively, as a result of validation experiments. EPS samples were characterized by Fourier-transform infrared spectroscopy and scanning electron microscope analyses. The optimization process caused more than a 50% increase in EPS and pullulan concentrations. This study showed that the color-variant A. pullulans AZ-6 strain could become a significant industrial strain if it is explored further in the future to produce pullulan on a larger scale. [ABSTRACT FROM AUTHOR]

The study aimed to perform a comprehensive in vitro and in vivo evaluation of a newly developed, patent-pending, powder-to-hydrogel, film-forming polymer complex base, which possesses tissue-protective and microbiome-supportive properties, and to compare its characteristics with poloxamer 407. The study used a combination of in vitro assays, including tissue viability and cell migration, and in vivo wound healing evaluations in male diabetic mice. Microbiome dynamics at wound sites were also analyzed. The in vitro assays demonstrated that the polymer complex base was non-cytotoxic and that it enhanced cell migration over poloxamer 407. In vivo, the polymer complex base demonstrated superior wound healing capabilities, particularly in combination with misoprostol and phenytoin, as evidenced by the reduced wound area and inflammation scores. Microbiome analysis revealed favorable shifts in bacterial populations associated with the polymer complex base-treated wounds. The polymer complex base demonstrates clinical significance in wound care, potentially offering improved healing, safety and microbiome support. Its transformative properties and efficacy in drug delivery make it a promising candidate for advanced wound care applications, particularly in chronic wound management. [ABSTRACT FROM AUTHOR]

Abstract Objectives The ascomycotic yeast-like fungus Aureobasidium exhibits the natural ability to synthesize several secondary metabolites, like polymalic acid, pullulan, or polyol lipids, with potential biotechnological applications. Combined with its polyextremotolerance, these properties make Aureobasidium a promising production host candidate. Hence, plenty of genomes of Aureobasidia have been sequenced recently. Here, we provide the annotated draft genome sequence of the polyol lipid-producing strain A. pullulans NRRL 62042. Data description The genome of A. pullulans NRRL 62042 was sequenced using Illumina NovaSeq 6000. Genome assembly revealed a genome size of 24.2 Mb divided into 39 scaffolds with a GC content of 50.1%. Genome annotation using Genemark v4.68 and GenDBE yielded 9,596 genes.

Abstract One major limitation of effective vaccine delivery is its dependency on a robust cold chain infrastructure. While Vesicular stomatitis virus (VSV) has been demonstrated to be an effective viral vaccine vector for diseases including Ebola, its −70 °C storage requirement is a significant limitation for accessing disadvantaged locations and populations. Previous work has shown thermal stabilization of viral vaccines with a combination of pullulan and trehalose (PT) dried films. To improve the thermal stability of VSV, we optimized PT formulation concentrations and components, as well as drying methodology with enhanced vacuum drying. When formulated in PT films, VSV can be stored for 32 weeks at 4 °C with less than 2 log PFU loss, at 25 °C with 2.5 log PFU loss, and at 37 °C with 3.1 log PFU loss. These results demonstrate a significant advancement in VSV thermal stabilization, decreasing the cold chain requirements for VSV vectored vaccines.

The quest for sustainable and functional food packaging materials has led researchers to explore biopolymers such as pullulan, which has emerged as a notable candidate for its excellent film-forming and anti-fogging properties. This study introduces an innovative anti-fog coating by combining pullulan with poly (acrylic acid sodium salt) to enhance the display of packaged food in high humidity environments without impairing the sealing performance of the packaging material—two critical factors in preserving food quality and consumers' acceptance. The research focused on varying the ratios of pullulan to poly (acrylic acid sodium salt) and investigating the performance of this formulation as an anti-fog coating on bioriented polypropylene (BOPP). Contact angle analysis showed a significant improvement in BOPP wettability after coating deposition, with water contact angle values ranging from ~60° to ~17° for formulations consisting only of poly (acrylic acid sodium salt) (P0) or pullulan (P100), respectively. Furthermore, seal strength evaluations demonstrated acceptable performance, with the optimal formulation (P50) achieving the highest sealing force (~2.7 N/2.5 cm) at higher temperatures (130 °C). These results highlight the exceptional potential of a pullulan-based coating as an alternative to conventional packaging materials, significantly enhancing anti-fogging performance. [ABSTRACT FROM AUTHOR]

During a survey of culturable microfungi from the bark of sugar maple (Acer saccharum), Atrocalyx glutinosus and Nigrograna rubescens, two novel species of Pleosporales (Dothideomycetes) were isolated from several locations in eastern Ontario, Canada. Formal species descriptions are presented based on unique colony phenotypes and micromorphological characteristics and supported using multi-locus molecular phylogenetic comparisons with similar species. Both A. glutinosus and N. rubescens produce pycnidial asexual morphs in culture. As their names imply, under specific culture conditions, A. glutinosus excretes large amounts of the glutinous polysaccharide pullulan and N. rubescens produces a dark red naphthoquinone pigment that diffuses in the culture medium. [ABSTRACT FROM AUTHOR]

Petroleum-based plastics have been widely used as packaging materials because of their low-cost availability and good mechanical properties. However, the use of plastics has become restricted as they are highly resistant to biodegradation, causing environmental problems. This work aimed to produce and characterize emulsified pullulan films incorporating geraniol for application as food packaging materials with potential to substitute the conventional plastics. When geraniol was incorporated in the films, they showed antimicrobial activity against Enterococcus faecalis ATCC 29212 (inhibition zone diameter = 15.19 ± 0.66 mm) and Pseudomonas aeruginosa ATCC 27853 (inhibition zone diameter = 10.99 ± 1.82 mm). Furthermore, scanning electron microscopy showed the inhibition of Enterococcus faecalis ATCC 29212 biofilms when they were directly formed on the emulsified pullulan films incorporating geraniol. The produced films also demonstrated high transparency (> 90%) and hydrophilic surfaces (water contact angle < 90°). This work demonstrated the viability of using geraniol to produce pullulan active films as new food packaging materials. [ABSTRACT FROM AUTHOR]

Due to its unique physicochemical properties, Pullulan is an exopolysaccharide with many applications in the food, biomedical, and pharmaceutical industries. Aiming to reduce its production cost, an interesting alternative is to consider other possibilities of raw materials, including the production of this biopolymer in a lignocellulosic biorefinery concept. Xylose is the main sugar of hemicellulosic hydrolysates obtained from different biomasses, and it is a sugar still not extensively exploited regarding its potential for pullulan production. This study aimed to evaluate the production of pullulan from sugarcane bagasse hemicellulosic hydrolysate by cultivating Aureobasidium pullulans ATCC 42023 in a bubble column reactor. The hemicellulosic hydrolysate was obtained through dilute acid treatment carried out in a stirred tank reactor before being detoxified to remove microbial growth inhibitors. The maximum concentration of 28.62 ± 1.43 g/L of pullulan was obtained after 120 h of fermentation in a bubble column reactor in batch mode. Analysis of spectroscopic properties through FTIR of the obtained pullulan revealed α-(1→6)-linked maltosyl units, similar to those of commercial samples of the biopolymer. XRD analysis showed that the prepared pullulan is amorphous, and a homogeneous morphology with a smooth surface of the pullulan was observed in SEM analysis. This study showed the potential of the production of pullulan from sugarcane bagasse hemicellulosic hydrolysate in a bubble column bioreactor, an alternative strategy for the industrial production of this biopolymer. [ABSTRACT FROM AUTHOR]

Pullulan is a polysaccharide that has attracted the attention of scientists in recent times as a former of edible films. On the other hand, its use for the preparation of hydrogels needs more study, as well as the formation of pullulan microcapsules as active ingredient release systems for the food industry. Due to the slow gelation kinetics of pullulan with sodium trimetaphosphate (STMP), capsules cannot be formed through the conventional method of dropping into a solution of the gelling agent, as with other polysaccharides, since the pullulan chains migrate to the medium before the capsules can form by gelation. Pullulan microcapsules have been obtained by using inverse water-in-oil emulsions as templates. The emulsion that acts as a template has been characterized by monitoring its stability and by optical microscopy, and the size of the emulsion droplets has been correlated with the size of the microcapsules obtained, demonstrating that it is a good technique for their production. Although some flocs of droplets form, these remain dispersed during the gelation process and two capsule size distributions are obtained: those of the non-flocculated droplets and the flocculated droplets. The microcapsules have been evaluated as vitamin C release systems, showing zero-order release kinetics for acidic pH and Fickian mechanism for neutral pH. On the other hand, the microcapsules offer good protection of vitamin C against oxidation during an evaluation period of 14 days. [ABSTRACT FROM AUTHOR]

As antibiotic resistance increasingly undermines traditional infection management strategies, there is a critical demand for innovative wound care solutions that address these emerging challenges. This study introduces a novel antibacterial wound dressing based on Cross-Linked Pullulan (Pul) and Polyhexamethylene Biguanide (PHMB) for enhanced wound management and infection control. The dressing's adsorption rate reached 200% of its original weight within 30 min, exceeded 300% after 5 h, and exhibited significant non-Newtonian fluid properties. The dressings were able to release the loaded medication completely within 20 min; additionally, the dressing demonstrated significant antibacterial activity against a broad spectrum of bacteria. Significantly, the therapeutic effects of the Pul-PHMB/GP dressing were evaluated in a mouse model. Compared to untreated wounds, wounds treated with Pul-PHMB/GP exhibited a significant gelation process within 5 min post-treatment and showed a significant increase in wound healing rate within 12 days. This powder preparation overcomes the limitations associated with liquid and gel dressings, notably in storage and precise application, preventing the premature expansion or dissolution often caused by PHMB in high-humidity environments. The powder form can transform into a gel upon contact with wound exudate, ensuring accurate coverage of irregular wounds, such as those from burns or pressure sores, and offers excellent chemical and physical stability in a dry state, which facilitates storage and transport. This makes the dressing particularly suitable for emergency medical care and precision therapy, significantly improving the efficiency and adaptability of wound treatment and providing robust support for clinical treatments and emergency responses. [ABSTRACT FROM AUTHOR]

A high-yielding microbial polysaccharide-producing strain, named RM1603, was isolated from rhizosphere soil and identified by morphological and phylogenetic analysis. The extracellular polysaccharides (EPS) were identified by thin-layer chromatography and infrared spectroscopy. The fermentation conditions were optimized by single factor experiments in shake flasks and a 5-L fermentor. The results of morphological and phylogenetic tree analysis showed that RM1603 was a strain of Aureobasidium pullulans. Its microbial polysaccharide was identified as pullulan, and the EPS production capacity reached 33.07 ± 1.03 g L−1 in shake flasks. The fermentation conditions were optimized in a 5-L fermentor, and were found to encompass an initial pH of 6.5, aeration rate of 2 vvm, rotor speed of 600 rpm, and inoculum size of 2 %. Under these conditions, the pullulan yield of RM1603 reached 62.52 ± 0.24 g L−1. Thus, this study contributes RM1603 as a new isolation with high-yielding pullulan and potential application value in biotechnology. [ABSTRACT FROM AUTHOR]

The development of hydrogels based on dextrans, pullulan and lentinan to be used in biomedical applications including tissue engineering is reported. Despite the fact that selected polysaccharides such as hyaluronic acid are well established, little is known, how these polysaccharides can be chemically modified to create hydrogels under controlled conditions. In this study we present a small library of chemically modified polysaccharides which are used for a divergent approach to achieve biomedical relevant hydrogels. In this case the crosslinking is based on thio ether formation between thiol modified donor and vinylsulfone or maleimide modified acceptor components. Successful synthesis of the linker systems and coupling at the polysaccharides, hydrogel formation takes place under physiological conditions. We extended the study by coupling small molecules like adhesion factors for increasing cell compatibility as well as a dye for further studies. The different hydrogels were studied to their rheological properties, water uptake, their permeability, biodegrability and their cytotoxicity. [ABSTRACT FROM AUTHOR]

Film dressings adhere well to the skin, even in the joints of the human body, and can serve as a wound barrier to promote wound healing. To improve water resistance of the film dressings, sodium trimetaphosphate (STMP) was selected to crosslink pullulan. Using glycerol as plasticizer and levofloxacin as antibacterial drug under alkaline condition, the pullulan composite films were prepared. Chemical structure, transparency, water resistance, water absorption, water vapor permeability, mechanical properties, antibacterial properties, and biocompatibility of the pullulan composite films were studied. Results showed that the composite films had high transparency and good water resistance, which not only facilitated the observation of wound healing, but also maintained its integrity during use, avoiding damage caused by the dissolution of exudate and delaying wound healing, when the addition amount (relative to the mass of pullulan, the same below) of STMP was 10%, glycerol was 30%, NaOH was 0.4%, and levofloxacin was 1%. At the same time, the composite films had appropriate water absorption, moisture permeability, and mechanical properties, which could absorb wound exudate, maintainappropriate humidity conditions for the wound healing to facilitate wound healing, and better fit the skin. Moreover, the composite films had 100% antibacterial activity against both Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli), and had good biocompatibility. The resultant pullulan composite films had potential application of wound healing. [ABSTRACT FROM AUTHOR]

Mucoadhesive microparticles for oromucosal drug delivery offer several advantages, including intimate contact with the mucosa, delivery to less accessible regions, extended residence time, sustained drug release, reduced irritation, and improved patient compliance. In this study, pullulan was used to prepare mucoadhesive spray-dried microparticles for delivering benzydamine hydrochloride (BZH) to oral mucosa. The BZH-pullulan spray-dried microparticles had a mean size of <25 μm with an angle of repose values between 25.8–36.6°. Pullulan markedly extended drug-release time to >180 min, ~9 times greater than the duration (i.e., 20 min) reportedly achieved by chitosan. Kinetic analysis showed the drug-release rate was concentration dependent and jointly controlled by drug diffusion and polymer chain relaxation. Further, pullulan was mucoadhesive and was able to retain up to 78.8% w/w of microencapsulated gold nanoparticle probes at the mucosal membrane. These data strongly suggest that BZH-pullulan microparticles have great potential for oromucosal drug delivery, by providing elongated residence time in situ and sustained drug release for the treatment of local diseases. [ABSTRACT FROM AUTHOR]

In the present study, Auricularia auricular polysaccharides (AAP) and Auricularia auricular proteins (AAPR) obtained from the waste products of Auricularia auricular were incorporated into pullulan (PUL) to obtain active packaging films/coatings. Results showed that incorporating AAP/AAPR into PUL-based films decreased their transparency, but increased the compactness, thermal stability, antioxidant, and antimicrobial properties. Adding 2% PUL films with 10%:10% of AAP/AAPR exhibiting good mechanical properties were applied to fresh-cut potatoes to avoid spoilage during eight days of storage, with significantly decreased in browning index, weight loss, microbial growth prevention and the total soluble solids was maintained. These results substantiated that pullulan containing AAP/AAPR as an active film/coating with antioxidant and antimicrobial properties has significant potential for maintaining safety and quality of fresh-cut potatoes and extending their shelf life. [ABSTRACT FROM AUTHOR]

We prepared tea tree essential oil microcapsules, and the microcapsules and pullulan were coated on kraft paper to prepare an antibacterial paper. The antibacterial activity, structural characterization, and thermal stability of the prepared microcapsules and packaging paper were then tested. We found that the retention rate of microcapsules reached 87.1% after a 70 min of high-temperature treatment. The minimum inhibitory concentrations of microcapsules to S. aureus and E. coli were 112 mg/mL and 224 mg/mL, and the bacteriostatic zones of the packaging paper to E. coli and S. aureus were 17.49 mm and 22.75 mm, respectively. The prepared microcapsules were irregular. The paper coating was formed via hydrogen bonding, which filled the pores of paper fibers. When compared with the base paper, the roughness of the paper was reduced to 7.16 nm (Rq) and 5.61 nm (Ra), and no thermal decomposition occurred at

Studies have demonstrated the protective effect of milk fat globule membrane (MFGM) on probiotics in harsh environments. However, currently, there are no reports on the encapsulation of probiotics using MFGM. In this study, MFGM and pullulan (PUL) polysaccharide fibers were prepared by electrostatic spinning and used to encapsulate probiotics, with whey protein isolates (WPI)/PUL as the control. The morphology, physical properties, mechanical properties, survival, and stability of the encapsulated Lacticaseibacillus rhamnosus GG (LGG) were studied. The results showed that the MFGM/PUL solution had significant effects on pH, viscosity, conductivity, and stability. Electrostatic spinning improved the mechanical properties and encapsulation ability of the polymer formed by MFGM/PUL. LGG encapsulated in MFGM/PUL nanofibers survived rate was higher than WPI/PUL nanofibers in mimic intestinal juice, which could be attributed to the phospholipid content contained in MFGM. These results demonstrate that MFGM is a promising material for probiotic encapsulation, providing an important basis for the potential use of MFGM/PUL nanofibers as a robust encapsulation matrix.

In recent years, the issue of electronic waste production has gained significant attention. To mitigate the environmental impact of e-waste, one approach under consideration involves the development of biodegradable electronic devices or devices that dissolve in the environment at the end of their life cycle. This study presents results related to the creation of a sensor that effectively addresses both criteria. The device was constructed using a composite material formed by impregnating a pullulan membrane (a biodegradable water-soluble biopolymer) with 1-Ethyl-3-Methylimidazolium tetrafluoroborate (a water-soluble ionic liquid) and coating the product with a conductive silver-based varnish. Capitalizing on the piezoionic effect, the device has demonstrated functionality as a vibration sensor with a sensitivity of approximately 5.5 × 10−5 V/mm and a resolution of about 1 mm. The novelty of this study lies in the unique combination of materials. Unlike the use of piezoelectric materials, this combination allows for the production of a device that does not require an external potential difference generator to function properly as a sensor. Furthermore, the combination of a biopolymer, such as pullulan, and an ionic liquid, both readily soluble in water, in creating an active electronic component represents an innovation in the field of vibration sensors. [ABSTRACT FROM AUTHOR]

In this study, polysaccharide-based nanofibrous fast dissolving oral films (FDOFs) were developed using pullulan (PUL) and xanthan gum (XG) via electrospinning. The edible, continuous, and bead-free nanofibers with average diameters ranging from 181.17 nm to 260.84 nm were prepared. The morphological, thermal, mechanical, and water-soluble properties of the nanofibrous FDOFs were characterized. For prospective future applications of the developed PUL/XG FDOFs, a model nutrient of vitamin C (VC) was encapsulated into the FDOFs. The success of VC encapsulation was confirmed by Fourier transform infrared spectroscopy. The encapsulation efficiency of VC was above 85% by ultraviolet-visible spectrophotometer. The amorphous structure of PUL/XG in the nanofibers film was demonstrated by X-ray diffractometer. In addition, the edible FDOFs could dissolve in water within 3 s. The nanofibers film we prepared could be used as nutrient or drug carriers and edible packaging film. [ABSTRACT FROM AUTHOR]

This study addresses the production enhancement of pullulan, an extracellular polysaccharide with various applications. Pullulan is primarily produced by Aureobasidium pullulans (A. pullulans), and genetic modification is commonly used to increase its yield. However, there is a need for a more efficient and safer method. To achieve this, we designed a high-throughput screening system utilizing a unique fluorescent protein specific to pullulan. Ultraviolet (UV) mutagenesis was applied to create a pool of mutant strains, and flow cytometry allowed for single-cell screening. Our approach yielded strain M1-B3, which exhibited a substantial increase in pullulan production from 26.5 g/L to 76.88 g/L. Additionally, the molecular weight of the produced pullulan significantly increased, expanding its potential commercial application. This study demonstrates an efficient and safe method to enhance pullulan production in A. pullulans. The UV mutagenesis and flow cytometry based on screening not only increased yield but also improved pullulan's molecular weight. The adaptability of this method to other polysaccharides and its potential for genomic analysis and broader applications make it a valuable tool in bioproduction. [ABSTRACT FROM AUTHOR]

The development of active edible coatings with improved mechanical and barrier properties is a huge challenge. In this study, active edible coatings for sliced cheese have been developed using pullulan (Pull) in combination with two different biopolymers, chitosan (CS) and gelatine (Gel), and a combination of hydrolats as a source of active compounds with antimicrobial effects. In comparison to the monolayer coating, the bilayer coating system demonstrates improved barrier and mechanical properties. A preliminary assessment of the antimicrobial effect of lemongrass and curry plant hydrolats has revealed that both hydrolats exhibited antimicrobial activity against the targeted bacterium Staphylococcus aureus, albeit at different levels. The obtained results suggest that a mixture of 1.56% lemongrass and 12.5% curry plant hydrolats yielded a lower fractional inhibitory concentration (FIC) value. Bilayer coating systems (Pull/CS and Pull/Gel) with an incorporated mixture of hydrolats have demonstrated effectiveness in both cases: artificial contamination before application of the coating system and after application of the coating system. In both contamination scenarios, the coating systems consistently effectively limited bacterial proliferation, indicating the antimicrobial effect of the hydrolat mixture in the coating layers. In the case of artificial contamination before applying the coating system, both coatings demonstrated antimicrobial effectiveness, but the formulation with chitosan had a biocide effect, while the other, with gelatine, had only a bacteriostatic effect in a long-term setting. In the second case, both Pull/CS and Pull/Gel coatings demonstrated effectiveness in inhibiting bacterial growth regardless of the moment of contamination of the sample; the Pull/CS coating showed slightly better antimicrobial activity, achieving complete elimination of bacteria earlier compared with the Pull/Gel coating system. [ABSTRACT FROM AUTHOR]

Purpose: This study aimed to prepare and evaluate mucoadhesive buccal patches of Sumatriptan Succinate (SUS) made from natural polymers to improve bioavailability, patient compliance, and dose-dependent adverse effects associated with currently available dosage forms. Materials and Methods: The solvent casting method was used with a 32 factorial design. The base polymer was water-soluble polysaccharide pullulan at 2% m/v. NaCMC, SA, PVA, and PVP-K30 was also added in various amounts. The polymer-plasticizer solution, SUS, and SS were combined and dried before being cut into 2 cm patches. The patch was water-resistant-backed and stored at room temperature in an airtight glass jar. Results: The patches had a mass uniformity of 61.75 mg to 84.18 mg and a film thickness of 0.2 mm to 0.5 mm. Some formulations demonstrated excellent folding endurance, with some exceeding 300 folds. The drug content ranged from 6.0 to 9.2 mg, with high drug loading efficiency in formulation SB29 (95%). The patches surface pH ranged from 6.0 to 7.2. At 120 min, formulation SA19 showed significant swelling (70±3%) but achieved maximum drug release (100.2%) in a shorter time. Permeation studies discovered a link between drug release and permeation. Various analyses and accelerated stability tests revealed no significant changes in physicochemical properties. Conclusion: This study demonstrated that the prepared mucoadhesive buccal patches of SUS might be useful in treating migraines, promising improved therapeutic outcomes, increased patient compliance, reduced side effects, and cost benefits. [ABSTRACT FROM AUTHOR]

An approach to the preparation of pullulan-graft-poly(2-methyl-2-oxazoline)s based on Cu-catalyzed azide–alkyne cycloaddition with polyoxazoline-azide was applied. All of the obtained polymers were characterized through classical molecular hydrodynamic methods and NMR. The formation of graft copolymers was accomplished by oxidative degradation of pullulan chains. Nevertheless, graft copolymers were obtained as uniform products with varied side chain lengths and degrees of substitution. [ABSTRACT FROM AUTHOR]

Pullulan, which is a microbial exopolysaccharide, has found widespread applications in foods, biomedicines, and cosmetics. Despite its versatility, most wild-type strains tend to yield low levels of pullulan production, and their mutants present genetic instability, achieving a limited increase in pullulan production. Therefore, mining new wild strains with robust pullulan-producing abilities remains an urgent concern. In this study, we found a novel strain, namely, Aureobasidium melanogenum ZH27, that had a remarkable pullulan-producing capacity and optimized its cultivation conditions using the one-factor-at-a-time method. To elucidate the reasons that drove the hyper-production of pullulan, we scrutinized changes in cell morphology and gene expressions. The results reveal that strain ZH27 achieved 115.4 ± 1.82 g/L pullulan with a productivity of 0.87 g/L/h during batch fermentation within 132 h under the optimized condition (OC). This pullulan titer increased by 105% compared with the initial condition (IC). Intriguingly, under the OC, swollen cells featuring 1–2 large vacuoles predominated during a rapid pullulan accumulation, while these swollen cells with one large vacuole and several smaller ones were prevalent under the IC. Moreover, the expressions of genes associated with pullulan accumulation and by-product synthesis were almost all upregulated. These findings suggest that swollen cells and large vacuoles may play pivotal roles in the high level of pullulan production, and the accumulation of by-products also potentially contributes to pullulan synthesis. This study provides a novel and promising candidate for industrial pullulan production. [ABSTRACT FROM AUTHOR]

Plant-based hard capsules have gained considerable attention because of their great properties. Accordingly, designing and developing of these kinds of capsules will be a difficult task. Herein, an innovative pullulan-based hard capsule formulation was prepared for the first time. A series of characterization approaches, including Fourier transform infrared, field emission scanning electron microscope, and rheology analysis, were utilized to figure out the straightforward preparation of a designed hard capsule. Many tests and experiments were performed to achieve the optimum capsule formulation. Based on the obtained results, specifications such as uniform downfall and non-desirable adhesion, and other ideal characteristics of the capsule display the critical function. The gelling promoter of divalent cationic salts is more beneficial than its single-valent counterparts. With respect to the key role of gelling promoter, the presence of chosen MgSO4.7H2O salt and the source of selected carrageenan are important parameters to achieve optimal formulation. Moreover, field emission scanning electron microscope images illustrate that the weight ratio of 3.5 (gelling agent to salt) displays uniform surface morphology without any impurities or other foreign materials. Likewise, the outcomes of the rheology test also illustrated that the weight ratio of 3.5 is preferable. Considering the different weight ratios, the benefits of a weight ratio of 3.5 outweigh the other investigated ratios. Overall, the current research addresses substantial information about developing pullulan-based hard capsules for target usage.

Abstract This article reports on the preparation of native/waxy corn starch (NCS/WCS)‐based films with the addition of pullulan (PUL). Our study investigated the effects of varying amounts of PUL and amylose contents on the structure, mechanical and physicochemical properties of corn starch films. Notably, it was observed that WCS films with low amylose content exhibited superior transparency, while NCS films with high amylose content demonstrated enhanced tensile strength (up to 7.35 ± 0.18 MPa). Fourier transform infrared spectroscopy (FTIR) analysis indicated that the addition of PUL did not change the molecular interactions within the corn starch films. The X‐ray diffraction (XRD) results supported that the WCS films were amorphous, while the NCS film exhibited B‐type and V‐type crystals. Moreover, an increase in PUL content led to a gradual reduction in the crystallinity of both WCS films and NCS films. The addition of PUL improved the mechanical properties and oxygen barrier characteristics of these films but had an adverse impact on their water vapor barrier properties. These findings offer valuable insights for the selection of additives for corn starch film, which can further enhance the practical application potential of corn starch films in food and other industries.

A microwave technique was used to prepare a superabsorbent polymer (SAP) by grafting two hydrophilic monomers onto a polysaccharide substrate. The monomers used were acrylic acid (AA) or acrylamide (AM) and were grafted onto a pullulan (PUL) substrate to form PUL-g-AA (SAP1) and PUL-g-AM (SAP2), respectively. The monomers (AM/AA) were grafted together onto a PUL substrate to form PUL-g-(AM/AA) (SAP3). Grafting parameters such as grafting efficiency with the percentage, the conversion of monomer into polymer, gel content, water retention, water adsorption capacity, and swelling kinetics were determined. Additionally, the effect of environmental pH (2, 4, 7, 9, and 12) and sodium dodecylbenzene sulfonate (SDBS) surfactant was evaluated, where 1, 2, 3, 4, and 5 mM of SDBS was added to form SAP4 to SAP8. The FTIR results show that AM was grafted onto PUL through an aliphatic C-N bond, while AA grafting occurred through a single C-C bond. The grafting efficiency with AM was higher than with AA, as well as showing a superior gel content. Water absorbance capacity and water retention increased with the grafting of AA and AM together for SAP3. The highest absorbent capacity, water retention, gel content, and grafting parameters values were obtained with a 3 mM SDBS content and a pH of 7. The swelling kinetics showed that the increases in the theoretical and experimental swelling equilibriums were 72% and 82%, respectively, for SAP6 compared to the values of these parameters for SAP3. The water absorption capacity of the hydrogel increases upon increasing the pH to 7 and then gradually decreases. XRD demonstrated the improved crystallinity and crystalline size of the hydrogel after grafting polymerization of AM/AA onto PUL, in addition to enhanced thermal stability. On the contrary, FE-SEM demonstrated that SDBS improves the porosity and pore size of the hydrogel surface with SAP6.

Pullulan, a natural polysaccharide with unique biocompatibility and biodegradability, has gained prominence in nanomedicine. Its application in nanoparticle drug delivery systems showcases its potential for precision medicine. Aim of Study: This scientific review aims to comprehensively discuss and summarize recent advancements in pullulan-based polymeric nanoparticles, focusing on their formulation, characterization, evaluation, and efficacy. Methodology: A search on Scopus, PubMed, and Google Scholar, using “Pullulan and Nanoparticle” as keywords, identified relevant articles in recent years. Results: The literature search highlighted a diverse range of studies on the pullulan-based polymeric nanoparticles, including the success of high-selectivity hybrid pullulan-based nanoparticles for efficient boron delivery in colon cancer as the active targeting nanoparticle, the specific and high-efficiency release profile of the development of hyalgan-coated pullulan-based nanoparticles, and the design of multifunctional microneedle patches that incorporated pullulan–collagen-based nanoparticle-loaded antimicrobials to accelerate wound healing. These studies collectively underscore the versatility and transformative potential of pullulan-based polymeric nanoparticles in addressing biomedical challenges. Conclusion: Pullulan-based polymeric nanoparticles are promising candidates for innovative drug delivery systems, with the potential to overcome the limitations associated with traditional delivery methods.

This article reports on the preparation of native/waxy corn starch (NCS/WCS)‐based films with the addition of pullulan (PUL). Our study investigated the effects of varying amounts of PUL and amylose contents on the structure, mechanical and physicochemical properties of corn starch films. Notably, it was observed that WCS films with low amylose content exhibited superior transparency, while NCS films with high amylose content demonstrated enhanced tensile strength (up to 7.35 ± 0.18 MPa). Fourier transform infrared spectroscopy (FTIR) analysis indicated that the addition of PUL did not change the molecular interactions within the corn starch films. The X‐ray diffraction (XRD) results supported that the WCS films were amorphous, while the NCS film exhibited B‐type and V‐type crystals. Moreover, an increase in PUL content led to a gradual reduction in the crystallinity of both WCS films and NCS films. The addition of PUL improved the mechanical properties and oxygen barrier characteristics of these films but had an adverse impact on their water vapor barrier properties. These findings offer valuable insights for the selection of additives for corn starch film, which can further enhance the practical application potential of corn starch films in food and other industries. [ABSTRACT FROM AUTHOR]

Pullulan is a polymer produced by Aureobasidium spp. The yield of pullulan production can be impacted by the cellular differentiation of Aureobasidium spp., which changes with alterations in the growth environment. To improve pullulan yield, identifying key factors that regulate cellular differentiation is crucial. In this study, the main form of pullulan synthesis in Aureobasidium pullulans NG was through swollen cells (SC). The results showed that citric acid (CA) can regulate the cellular differentiation of Aureobasidium pullulans NG by accumulating higher levels of CA in the cells to maintain growth in SC form and increase pullulan production. The addition of 1.0% CA to Aureobasidium pullulans NG for 96 h resulted in a significant increase in pullulan production, producing 18.32 g/l compared to the control group which produced 10.23 g/l. Our findings suggest that controlling cellular differentiation using CA is a promising approach for enhancing pullulan production in Aureobasidium pullulans. Key points: • The regulation of cell differentiation in Aureobasidium pullulans NG is demonstrated to be influenced by citric acid. • Intracellular citric acid levels in Aureobasidium pullulans NG have been shown to support the growth of swollen cells. • Citric acid has been found to increase pullulan production in Aureobasidium pullulans NG. [ABSTRACT FROM AUTHOR]

More sustainable and smart cosmeceuticals and nutraceuticals are necessary due to the ecological transition. In this study, a pullulan-based water solution containing chitin nanofibril–nano-lignin (CN-LG) complexes that encapsulate fish collagen polypeptide, allantoin and nicotinamide was electrospun onto a nonwoven substrate made of bamboo fibers to obtain a smart nanostructured bilayer system for releasing active molecules onto the skin or other body tissues. Infrared spectroscopy was used to characterize the composition of the bilayer system before and after rapid washing of the sample with distilled water and liquids mimicking physiological fluids. The viability of keratinocytes was studied as well as the antioxidant activity, protective activity towards UV light, metalloproteinase release of aged fibroblasts and the inhibitor activity against collagen degradation. Immunomodulatory tests were performed to investigate the anti-inflammatory activity of the bilayer system as well as its indirect antimicrobial activity. The results indicate that the bilayer system can be used in the production of innovative sustainable cosmeceuticals. In general, the adopted strategy can be extended to several smart treatments for fast release that can be commercialized as solid products, thus avoiding the use of preservatives and water. [ABSTRACT FROM AUTHOR]

Introduction: Guided bone regeneration (GBR) procedures require selecting suitable membranes for oral surgery. Pullulan and/or dextran-based polysaccharide materials have shown encouraging results in bone regeneration as bone substitutes but have not been used to produce barrier membranes. The present study aimed to develop and characterize pullulan/dextran-derived membranes for GBR. Materials and methods: Two pullulan/dextran-based membranes, containing or not hydroxyapatite (HA) particles, were developed. In vitro, cytotoxicity evaluation was performed using human bone marrow mesenchymal stem cells (hBMSCs). Biocompatibility was assessed on rats in a subcutaneous model for up to 16 weeks. In vivo, rat femoral defects were created on 36 rats to compare the two pullulan/dextran-based membranes with a commercial collagen membrane (Bio-Gide®). Bone repair was assessed radiologically and histologically. Results: Both polysaccharide membranes demonstrated cytocompatibility and biocompatibility. Micro-computed tomography (micro-CT) analyses at two weeks revealed that the HA-containing membrane promoted a significant increase in bone formation compared to Bio-Gide®. At one month, similar effects were observed among the three membranes in terms of bone regeneration. Conclusion: The developed pullulan/dextran-based membranes evidenced biocompatibility without interfering with bone regeneration and maturation. The HA-containing membrane, which facilitated early bone regeneration and offered adequate mechanical support, showed promising potential for GBR procedures. [ABSTRACT FROM AUTHOR]

Polyaniline (PANI), a conducting polymer, has great interest for a large number of applications. However, poor processability and mechanical properties limits its usage and many methods like blending, grafting etc. are used to overcome this disadvantage. We have carried out grafting of PANI onto pullulan (PULL) via chemical oxidative polymerization technique. The percentage of grafting is favored by increasing concentration of aniline monomer. The formation of PANI is confirmed through UV–Vis spectroscopic studies. The possible grafting mechanism is studied using Fourier transform infrared spectroscopy and validated by Hartree‐Fock density functional theory (HF‐DFT) calculations. Further, thermal properties of grafted polymers are studied using differential scanning calorimetry and thermo gravimetric analysis. Using FESEM and x‐ray diffraction, structural properties of graft polymer were studied. DC electrical conductivity of grafted polymer is measured from I‐V characteristics, shows a significant conductivity which is the highlight of this work. [ABSTRACT FROM AUTHOR]

Colon cancer is the second most invasive cancer and fourth most common malignant neoplasm worldwide. Targeted oral colonic drug delivery systems have attracted considerable attention in the treatment of colon cancer due to their superior properties. However, the delivery of drugs safely and effectively to the target site of the colon tumor is a hindrance due to the complexity of the gastrointestinal structure. Herein, to achieve an effective delivery system specifically targeting the colon, we have taken concern by using natural polymer such as pullulan, signifying its flexibility and relevance in biomaterials science to design antineoplastic approaches. Here, we summarize the physicochemical properties, different pullulan derivatives and their biomedical application, several colon cancer‐related treatment, and pullulan and its derivatives‐based delivery systems towards colonic tissue. [ABSTRACT FROM AUTHOR]

A nutrient medium was selected for the efficient production of exopolysaccharide (EPS) by A. pullulans C7 strain. The production of pullulan polysaccharide was evaluated on nutrient media with traditional carbon sources and cheap substrates that were plant wastes. For maximum EPS accumulation, we proposed an optimized Czapek-Dox medium with glucose as a carbon source, sodium nitrate as a nitrogen source, and C/N=232:1 ratio (EPS yield 12.79±0.64 g/l). Medium with grape pomace 5% (EPS yield was 15.08±0.34 g/l) and medium with topinambour tuber hydrolysate 5% (EPS yield was 14.44±0.21 g/l) was proposed as a cheap substrate. Edible films with antimicrobial activity were obtained on the basis of the isolated polysaccharide. The antibacterial activity of films against Escherichia coli 603 and Staphylococcus aureus ST228 was shown when essential oils of rosemary (zones of growth inhibition from 8.41±0.71 to 9.98±0.32 mm) and oregano (zones of growth inhibition from 8.09±0.51 to 9.54±0.24 mm) were added to pullulan. The addition of xanthan gum and glycerol to the films increased their strength and elasticity. The infrared spectrum of the pullulan film showed absorption bands characteristic of polysaccharide structures. [ABSTRACT FROM AUTHOR]

Collagen electrospun fibers are promising materials for food packaging and tissue engineering. The conventional electrospinning of collagen, however, is usually carried out by dissolving it in organic reagents, which are toxic. In this study, collagen/pullulan (COL/PUL) ultra-thin fibers were prepared by electrospinning using acetic acid as a solvent. Compared to the conventional preparation method, the proposed method is safe and does not produce toxic solvent residues. The introduction of PUL increased the degree of molecular entanglement in the solution, so the viscosity of the COL/PUL electrospun solution increased from 0.50 ± 0.01 Pa∙s to 4.40 ± 0.08 Pa∙s, and the electrical conductivity decreased from 1954.00 ± 1.00 mS/cm to 1372.33 ± 0.58 mS/cm. Scanning electron microscopy analysis confirmed that PUL improved the spinnability of COL, and smooth, defect-free COL/PUL ultra-thin fibers with diameters of 215.32 ± 40.56 nm and 240.97 ± 53.93 nm were successfully prepared at a viscosity of greater than 1.18 Pa∙s. As the proportion of PUL increased, intramolecular hydrogen bonds became the dominant interaction between COL and PUL. The intermolecular hydrogen bonding content decreased from 52.05 % to 36.45 %, and the intramolecular hydrogen bonding content increased from 46.11 % to 62.95 %. The COL was gradually unfolded, the content of α-helices decreased from 33.57 % to 25.91 % and the random coils increased from 34.22 % to 40.09 %. More than 36 % of the triple helix fraction of COL was retained by the COL/PUL ultra-thin fibers, whereas only 16 % of the triple helix fraction of COL was retained by the COL nanofibers prepared with 2.2.2-trifluoroethanol. These results could serve as a reference for the development of green food COL-based fibers.