Keratin Proteins Research Articles

Application of 4 × 44 Oligo Microarray to Transcriptomic Analysis of Immune Response in Rainbow Trout Infected with Aeromonas salmonicida

Rainbow trout, one of the most economically important aquaculture fish species worldwide, is affected by the pathogenic bacteria A. salmonicida, which causes furunculosis outbreaks, leading to huge economic losses. In this study, an oligonucleotide microarray was applied to identify transcriptional changes in the skin of rainbow trout individuals in response to a bacterial infection. Overall, 656 and 434 differentially expressed genes (DEGs) were identified at 2 and 6 days after a bacterial challenge (dpi), respectively. A comparison of moribund (2 dpi) and survivor fish (6 dpi) revealed 169 DEGs. Between these were many genes involved in immune response, including lysozymes, pattern recognition receptors (c-type lectins), antimicrobial peptides (cathelicidin and hepcidin), acute-phase proteins (serum amyloids and haptoglobin), complement cascade proteins (c3, c4, c6 and c7), interleukins (il11 and il1b) and chemokines (ccl19 and cxcl8). Alterations of leptin, eicosanoids and prostaglandins have been found, which suggest metabolic remodeling in conjunction with immune response. Further, the regulation of programmed cell death genes (caspase 8, bcl2 apoptosis regulator, nfkb inhibitor alpha and heme oxygenase) and structural proteins (collagens, myosins, keratins and metalloproteinases) was observed. This study provides, for the first time, a gene expression analysis of rainbow trout skin in response to A. salmonicida infection, revealing the complexity of defense strategies in response to furunculosis.

Extraction and Characterization of Keratin from Waste Broiler Chicken Feathers

The current study was to extract the keratin protein from waste broiler chicken feathers. Due to their high keratin content, chicken feathers offer a valuable source of protein. The two steps involved are treating sodium sulphide (Na2S) and then extracting the protein from the mixture. After the feathers have been broken down using reducing agents, dilute hydrochloric acid (HCl) is added to the solution to precipitate the protein. The precipitated protein was repeatedly rinsed with distilled water and dissolved in a sodium hydroxide (NaOH) solution to form a solution. The results indicate that a chicken feather has unique features. Keratin obtained from broiler chicken feathers had a yield of 81.1%, which is relatively high in cases of waste raw material conversion into value-added products. The prepared keratin was characterized by Fourier-transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), and thermogravimetric analysis (TGA) to investigate the chemical composition, thermal properties, crystallinity properties, and also physico-mechanical properties, including density. These findings suggest that chicken feathers can be used to extract protein.

A small molecule chaperone rescues keratin-8 mediated trafficking of misfolded podocin to correct genetic Nephrotic Syndrome

Podocin is a key membrane scaffolding protein of the kidney podocyte essential for intact glomerular filtration. Mutations in NPHS2, the podocin-encoding gene, represent the commonest form of inherited nephrotic syndrome (NS), with early, intractable kidney failure. The most frequent podocin gene mutation in European children is R138Q, causing retention of the misfolded protein in the endoplasmic reticulum. Here, we provide evidence that podocin R138Q (but not wild-type podocin) complexes with the intermediate filament protein keratin 8 (K8) thereby preventing its correct trafficking to the plasma membrane. We have also identified a small molecule (c407), a compound that corrects the Cystic Fibrosis Transmembrane Conductance Regulator protein defect, that interrupts this complex and rescues mutant protein mistrafficking. This results in both the correct localization of podocin at the plasma membrane and functional rescue in both human patient R138Q mutant podocyte cell lines, and in a mouse inducible knock-in model of the R138Q mutation. Importantly, complete rescue of proteinuria and histological changes was seen when c407 was administered both via osmotic minipumps or delivered orally prior to induction of disease or crucially via osmotic minipump two weeks after disease induction. Thus, our data constitute a therapeutic option for patients with NS bearing a podocin mutation, with implications for other misfolding protein disorders. Further studies are necessary to confirm our findings.

Structure prediction, docking studies and molecular cloning of novel Pichia kudriavzevii YK46 metalloprotease (MetPr) for improvement of feather waste biodegradation

This study addresses the environmental risks associated with the accumulation of keratin waste from poultry, which is resistant to conventional protein degradation methods. To tackle this issue, microbial keratinases have emerged as promising tools for transforming resilient keratin materials into valuable products. We focus on the Metalloprotease (MetPr) gene isolated from novel Pichia kudriavzevii YK46, sequenced, and deposited in the NCBI GenBank database with the accession number OQ511281. The MetPr gene encodes a protein consisting of 557 amino acids and demonstrates a keratinase activity of 164.04 U/ml. The 3D structure of the protein was validated using Ramachandran's plot, revealing that 93% and 97.26% of the 557 residues were situated within the most favoured region for the MetPr proteins of template Pichia kudriavzevii strain 129 and Pichia kudriavzevii YK46, respectively. Computational analyses were employed to determine the binding affinities between the deduced protein and beta keratin. Molecular docking studies elucidated the optimal binding affinities between the metalloprotease (MetPr) and beta-keratin, yielding values of − 260.75 kcal/mol and − 257.02 kcal/mol for the template strains Pichia kudriavzevii strain 129 and Pichia kudriavzevii YK46, respectively. Subsequent molecular cloning and expression of the MetPr gene in E. coli DH5α led to a significantly higher keratinase activity of 281 ± 12.34 U/ml. These findings provide valuable insights into the potential of the MetPr gene and its encoded protein for keratin waste biotransformation, with implications for addressing environmental concerns related to keratinous waste accumulation.

A Review on the Prospective Applications of Fungal Keratinases

Keratin is the most abundant insoluble protein in the epithelial cells of vertebrates and represents the major constituents of skin, hair, feathers, wool, and nails. It is among the hardest-to-degrade animal protein, the major component in poultry feathers. Keratinases are exciting proteolytic enzymes that display the capability to degrade the insoluble protein keratin, recalcitrance to common proteolytic enzymes like pepsin, trypsin, and papain. Keratinases are versatile enzymes gaining momentum in the field of Biotechnology with potential applications. As the keratin is too hard to degrade, substrates like hair, hoof, horn, feather nail etc. can be treated effectively with keratinases, which is considered as the remarkable application of the enzyme. The poultry feather degradation using keratinase improve the nutritional value of feather meal and can be applied for the production of rare amino acids like proline, cysteine, serine, and methionine. The potential use of these enzymes have been associated with cosmetic industry and acne treatment. The other applications of keratinase include the production of biofuel, detergent, films, coatings, glues and pesticidal toxins, degumming of silk, scale removal from wool fibre, degradation of prions etc. Keratinases are mainly produced by different microorganisms like fungi and bacteria which are considered as biodegradable, eco-friendly enzymes without any harmful effects like that of the chemical counterparts. As the separation of bacterial keratinase is a cost intensive process compared to fungal keratinase, the review is focussed on isolation, purification and prospective applications of fungal keratinase.

Application and Future Perspectives of Keratin Protein Extracted from Waste Chicken Feather: A Review

In the present review, we have focused on the utilization of keratin from poultry bio-mass. Keratin is a tough, fibrous protein that is mostly found. There are millions of tonnes of keratin-containing biomass produced by the food sector, particularly the meat market, slaughterhouses and wool industry. These proteins which make up the byproduct of keratin contain between 15-18% of Nitrogen, 2-5% of sulphur, 1.20% of Fat and 90% of Protein. It is an eco-friendly and economical method for the production of uniaxially and randomly oriented polymeric products. The chemical and mechanical techniques can be utilized right away to make a wide range of cosmetics, lotions, shampoos, hair conditioners and biomedical items. The natural protein is better suited for usage on or application to human hair and skin. In the present study, various sources of keratin, purification and separation of keratin from organic waste have industrial application and their future perspectives have all been discussed.

Substrate Specificity of Bacillus megaterium UСM B-5710 Keratinase

The specifics of the processing of livestock and poultry products is that in the process of obtaining the main marketable products, about half the feedstock at various stages of the technological process turns into waste that pollutes the environment. These by-products contain large amounts of the hard-to-digest keratin protein. The use of specific enzymes capable of degrading this protein helps not only to reduce the negative anthropogenic impact on nature but also to obtain valuable hydrolysates that can be used as a fertilizer for plants or a feed additive. The aim of this work was to study the ability of Bacillus megaterium UCM B-5710 to split various keratin-containing substrates: black and white chicken feathers, white turkey feathers, parrot feathers of various colors, sheep wool, pig bristles, and baby hair and nails. Methods. The culture was grown under conditions of submerged cultivation at 40 °C, with a nutrient medium stirring rate of 201 rpm for 6 days. For growth, a basic nutrient medium containing 0.5% defatted chicken feathers or other keratin-containing substrates as sole sources of carbon and nitrogen were used. Keratinase activity was assessed by UV absorption at 280 nm of hydrolysis products of keratin-containing raw materials. Protein was determined by the Lowry method, caseinolytic (total proteolytic) activity was determined by the Anson method modified by Petrova, and amino acid content was determined by the ninhydrin method. The degree of hydrolysis of the substrates was evaluated by the ratio of the initial and final weight of the substrate. Results. It was shown that the synthesis of keratinase by the culture of B. megaterium UCM B-5710 begins from the 6th hour of cultivation. The level of protein and proteolytic activity and the content of amino acids increased throughout the entire period of culture growth. The supernatant of the culture liquid of B. megaterium UCM B-5710 was most effective in splitting white chicken’s and turkey’s feathers, a little slower — feathers of black chicken and blue parrots, as well as wool of white sheep. According to the degree of splitting, the substrates used can be arranged in the following order: white turkey feathers > white chicken feathers > black chicken feathers > blue parrot feathers > white sheep wool > baby nails > pig bristle > baby hair. The study of the effect of feather color on the resistance to decomposition showed that black, blue, and red feathers are more resistant, which coincides with the literature data. Conclusions. B. megaterium UCM B-5710 produces keratinase capable of splitting both α- and β-keratins, however, with different efficiencies and rates.

Changes to insulin sensitivity in glucose clearance systems and redox following dietary supplementation with a novel cysteine-rich protein: A pilot randomized controlled trial in humans with type-2 diabetes

We recently developed a novel keratin-derived protein (KDP) rich in cysteine, glycine, and arginine, with the potential to alter tissue redox status and insulin sensitivity. The KDP was tested in 35 human adults with type-2 diabetes mellitus (T2DM) in a 14-wk randomised controlled pilot trial comprising three 2×20 g supplemental protein/day arms: KDP-whey (KDPWHE), whey (WHEY), non-protein isocaloric control (CON), with standardised exercise. Outcomes were measured morning fasted and following insulin-stimulation (80 mU/m2/min hyperinsulinaemic-isoglycaemic clamp). With KDPWHE supplementation there was good and very-good evidence for moderate-sized increases in insulin-stimulated glucose clearance rate (GCR; 26%; 90% confidence limits, CL 2%, 49%) and skeletal-muscle microvascular blood flow (46%; 16%, 83%), respectively, and good evidence for increased insulin-stimulated sarcoplasmic GLUT4 translocation (18%; 0%, 39%) vs CON. In contrast, WHEY did not effect GCR (-2%; -25%, 21%) and attenuated HbA1c lowering (14%; 5%, 24%) vs CON. KDPWHE effects on basal glutathione in erythrocytes and skeletal muscle were unclear, but in muscle there was very-good evidence for large increases in oxidised peroxiredoxin isoform 2 (oxiPRX2) (19%; 2.2%, 35%) and good evidence for lower GPx1 concentrations (-40%; -4.3%, -63%) vs CON; insulin stimulation, however, attenuated the basal oxiPRX2 response (4%; -16%, 24%), and increased GPx1 (39%; -5%, 101%) and SOD1 (26%; -3%, 60%) protein expression. Effects of KDPWHE on oxiPRX3 and NRF2 content, phosphorylation of capillary eNOS and insulin-signalling proteins upstream of GLUT4 translocation AktSer437 and AS160Thr642 were inconclusive, but there was good evidence for increased IRSSer312 (41%; 3%, 95%), insulin-stimulated NFκB-DNA binding (46%; 3.4%, 105%), and basal PAK-1Thr423/2Thr402 phosphorylation (143%; 66%, 257%) vs WHEY. Our findings provide good evidence to suggest that dietary supplementation with a novel edible keratin protein in humans with T2DM may increase glucose clearance and modify skeletal-muscle tissue redox and insulin sensitivity within systems involving peroxiredoxins, antioxidant expression, and glucose uptake.

Research Progress in Composition, Classification and Influencing Factors of Hair

Human hair is a natural fiber with keratin and keratin-related proteins as a main component. Externally, hair is a thin, pliable tube of dead, fully keratinized epithelial cells, while inside the skin, it is part of a single living hair follicle. In addition to a large amount of protein, hair also contains low levels of lipids and pigments. The hair also has its own ecological balance system. Although the lipid content in hair is much lower than the protein content, it plays an important role in hair and influences hair texture to some extent. For example, prevent hair breakage and thinning; To act as a barrier to moisture loss; and improve the gloss, elasticity and tensile strength of the hair stem. Environment, hair dressing and daily care can all cause hair damage to varying degrees. Factors such as ultraviolet light and chemicals can make hair dry, rough, dull, stiff and brittle by destroying the proteins, pigment sand lipid sin the hair. In this paper, the composition, classification and influencing factors of hair were reviewed. The composition and function of lipids in hair were investigated emphatically. It aims to improve people's understanding of the physical and chemical properties and structure of hair. It provides reference value for the research and development of hair cosmetics for different hair states and washing purposes. The function of hair lipids on hair is not accurate. In addition, there is still some controversy on the existing role of hair lipids, which needs further research.

Exhaustive Review on Nail Lacquer in the Treatment of Nail Diseases

The delivery of drugs through the transungual route is necessary because many nail diseases such as onychomycosis, psoriasis, paronychia, and onycholysis require topical drug delivery through the nail plate to avoid the systemic side effects of drugs first-pass hepatic metabolism and to provide direct benefits at the site of disease. However, topical delivery is associated with the main drawback of poor permeability of drugs through the nail plate. The nail plate contains tight networks of cross-linked protein (keratin) that provide hardness to the nail. Successful permeation of drugs through this nail barrier can be achieved by mechanical or chemical approaches. The chemical methods (penetration enhancers) are mainly associated with the disruption of nail integrity. Nail lacquers are the most popular dosage form from the formulation viewpoint. As the solvent evaporates, lacquers create polymeric drug-loaded films on the nail tissue that have a high concentration of the drug and allow for prolonged contact with the tissue. The present article focuses on the emergence of nail lacquers as prospective transungual delivery systems for the treatment of various mild to moderate nail diseases.

Sustainable Valorization of Chicken Feathers and Grocery Waste as Organic Fertilizer and its Impact on Yield and Quality of Spinach (Spinacia oleracea) Plant

ABSTRACT Overproduction of chicken feathers is responsible for serious environmental problems worldwide. The composition of feathers is keratin protein, which is resistant and difficult to monomerize. Utilization of feather waste for biogas and organic fertilizer production can help combat environmental threat and address energy and food shortage crisis. In this study, chicken feathers were hydrolyzed using the native isolated keratinolytic bacterial strain Pseudomonas aeruginosa-C1M. Reactor digestate, a by-product of the production of biogas from hydrolyzate and grocery waste, has been shown to be used as organic fertilizer. A greenhouse study was conducted to investigate the comparative effectiveness of reactor digestate and feather hydrolyzate on the growth of spinach plants. A remarkable increase in agronomic parameters was observed when applying keratin hydrolyzate based fertilizers. Plants treated with dry hydrolyzate showed a 50% increase in leaf width. The plant height, the number of leaves per plant, the root length, the chlorophyll content, the protein content, the biomass and the nutrient content of the spinach plant were also significantly improved by using fertilizers based on keratin hydrolyzate. However, the results using the Pseudomonas aeruginosa-C1M-derived hydrolyzate were more promising and therefore suggest its application as a potential biostimulant in the agricultural industry.

Physicochemical study of keratin containing impurities from human nails

The purpose of this research to study of physicochemical properties of keratin from human nails which still contains impurities. The method in this study, keratin from human nail waste was dissolved using NaOH solvent at room temperature, then neutralized to pH 7 using glacial acetic acid. On this report have studied the analysis of functional groups of keratin with impurities, viscosity, melting point, solubility and diffractogram pattern using XRD. The FTIR spectrum shows the transmittance in the wave number 1750-1200 cm−1 which is the characterization region of keratin protein. The intrinsic viscosity of nail keratin was obtained at 0.105 g/dL or 10 g/ml. The melting point of nail keratin was obtained at 210 °C. Keratin solubility test using HCl and NaOH 0.1 M and 1 M respectively, showed that keratin was soluble in strong base NaOH with high concentration. The results of XRD analysis, in a diffractogram pattern without a dominant peak and tend to widen, keratin from human nails is included in the amorphous structure type.

The Novelty of Amino Acid Production From Feathers Degradation Em-ploying Keratinase from Streptomyces Venezuelae AZ15

Keratinases are enzymes belonging to the serine hydrolases group which are capable of degradation of keratin, an insoluble and fibrous structural protein widely cross-linked with hydrogen, disulfide, and hydrophobic bonds. Attempts to find new sources of enzymes and amino acids for fundamental knowledge of enzyme evolution, structure‐function relationships, catalysis mechanisms of enzymes, and even for the identification of novel protein folds. In this study, seventy-nine samples were collected from different places in the University of Baghdad, and the best isolate for amino acid production by feathers degradation was by using Streptomyces venezuelae AZ15. The best fermentation system and the optimum culture conditions for amino acid production from feathers as substrate utilizing S. venezuelae AZ15 were elucidated. Using feathers as a substrate, the results showed that the submerged fermentation (SmF) system was the best to give the highest yield of amino acid. Furthermore, the highest yield of total amino acid was gained in feather broth supplemented with 2% glucose and 0.5 % ammonium nitrate, with a pH of 6.0 at 37 °C for 8 days.

Bifidobacterium longum 68S mediated gut-skin axis homeostasis improved skin barrier damage in aging mice

BackgroundBifidobacterium as probiotics, play important roles in skin status, while the potential mechanisms interaction remains unknown. The study further explored the potential mechanism of B. longum 68S in ameliorating skin barrier damage from the perspective of the gut-skin axis in aging mice. MethodsB. longum 68S supplied natural aging mouse model and fecal microbiota transplantation (FMT) experiment proves the key role of intestinal microbiota in B. longum 68S up-regulating the production of ceramide synthesis key enzyme (SPT1) and ceramide level and improving skin barrier damage. Moreover, B. longum 68S supplied SPT1 gene deletion mouse model to investigate the mechanism of B. longum 68S on improving skin barrier damage. ResultsTranscriptome analysis and 16S rRNA high-throughput pyrosequencing demonstrated that aging mice exhibited skin barrier dysfunction and intestinal dysbiosis. Meanwhile, aging mice exhibited an up-regulation in the trans epidermal water loss (TEWL) and a down-regulation in the level of SPT1, ceramide and skin barrier-related proteins (Loricrin, Keratin 10 and Desmoglein 1). Similarity, the FMT from aging mice to normal mice and SPT1 gene deletion mice could rebuild skin barrier damage and B. longum 68S supplementation exerted a positive effect on it. Further, B. longum 68S-mediated SPT1-derived ceramide production prevented impaired ceramide synthesis-induced endoplasmic reticulum stress and apoptotic response, ultimately improving skin barrier damage in vitro. ConclusionEmerging anti-aging therapies are necessary given the poor safety profiles of current pharmaceutical drugs. B. longum 68S may be better alternatives, considering the association between the gut microbiota and healthy aging. The findings suggested that B. longum 68S-mediated gut-skin axis homeostasis, thereby exhibiting an anti-aging effect and facilitate a better understanding of the mechanisms governing the various beneficial effects of B. longum 68S.

Process optimization, antioxidant, antibacterial, and drug adjuvant properties of bioactive keratin microparticles derived from porcupine (Hystrix indica) quills.

A structural protein called keratin is often employed in the medical industry to create medication carriers. Process improvement, antioxidant, antibacterial, and adjuvant drug studies of synthetic bioactive keratin microparticles made from lipids and keratin derived from porcupine (Hystrix indica) quills are the main objectives of this study. After coating the keratin microparticles with lipids which were obtained from the same porcupine quills, the bioactive keratin microparticles were produced. The response surface technique was applied to optimize the conditions for extraction of the keratin protein and sizing of the keratin microparticles. An infrared spectroscopy was used to analyze the chemical shifts in compositions of keratin microparticles while the optical microscopy was used to measure the size of the keratin microparticles. The results of this work revealed that a yield 27.36 to 42.25% of the keratin protein could be obtained from porcupine quills. The keratin microparticles were sized between 60.65 and 118.87 µm. Through response surface optimization, mercaptoethanol and urea were shown to be the main variables which positively affected the yield and the size of the keratin protein. The lipid stacking on the keratin microparticles' surface was confirmed by infrared spectroscopy. The 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulphonate) assay confirmed the keratin microparticle's antioxidant activity of 29.83%. Compared to lipid alone, the antibacterial properties of the keratin microparticles against Escherichia coli-a gram-negative-and Staphylococcus aureus-a gram-positive-bacteria enhanced by up to 55% following the coating of the microparticles with the lipids. The pharmacological action against these bacterial species was further improved by the lipid-loaded erythromycin that was carried on the surface of keratin microparticles. This work has demonstrated the design and uses of the keratin microparticles obtained from porcupine quills for clinical applications.

Keratin 19 binds and regulates cytoplasmic HNRNPK mRNA targets in triple-negative breast cancer.

Heterogeneous nuclear ribonucleoprotein K (HNRNPK) regulates pre-mRNA processing and long non-coding RNA localization in the nucleus. It was previously shown that shuttling of HNRNPK to the cytoplasm promotes cell proliferation and cancer metastasis. However, the mechanism of HNRNPK cytoplasmic localization, its cytoplasmic RNA ligands, and impact on post-transcriptional gene regulation remain uncharacterized. Here we show that the intermediate filament protein Keratin 19 (K19) directly interacts with HNRNPK and sequesters it in the cytoplasm. Correspondingly, in K19 knockout breast cancer cells, HNRNPK does not localize in the cytoplasm, resulting in reduced cell proliferation. We comprehensively mapped HNRNPK binding sites on mRNAs and showed that, in the cytoplasm, K19-mediated HNRNPK-retention increases the abundance of target mRNAs bound to the 3' untranslated region (3'UTR) at the expected cytidine-rich (C-rich) sequence elements. Furthermore, these mRNAs protected by HNRNPK in the cytoplasm are typically involved in cancer progression and include the p53 signaling pathway that is dysregulated upon HNRNPK knockdown (HNRNPK KD) or K19 knockout (KRT19 KO). This study identifies how a cytoskeletal protein can directly regulate gene expression by controlling the subcellular localization of RNA-binding proteins to support pathways involved in cancer progression.

Deacetylation via SIRT2 prevents keratin-mutation-associated injury and keratin aggregation.

Keratin (K) and other intermediate filament (IF) protein mutations at conserved arginines disrupt keratin filaments into aggregates and cause human epidermolysis bullosa simplex (EBS; K14-R125C) or predispose to mouse liver injury (K18-R90C). The challenge for more than 70 IF-associated diseases is the lack of clinically utilized IF-targeted therapies. We used high-throughput drug screening to identify compounds that normalized mutation-triggered keratin filament disruption. Parthenolide, a plant sesquiterpene lactone, dramatically reversed keratin filament disruption and protected cells and mice expressing K18-R90C from apoptosis. K18-R90C became hyperacetylated compared with K18-WT and treatment with parthenolide normalized K18 acetylation. Parthenolide upregulated the NAD-dependent SIRT2, and increased SIRT2-keratin association. SIRT2 knockdown or pharmacologic inhibition blocked the parthenolide effect, while site-specific Lys-to-Arg mutation of keratin acetylation sites normalized K18-R90C filaments. Treatment of K18-R90C-expressing cells and mice with nicotinamide mononucleotide had a parthenolide-like protective effect. In 2 human K18 variants that associate with human fatal drug-induced liver injury, parthenolide protected K18-D89H- but not K8-K393R-induced filament disruption and cell death. Importantly, parthenolide normalized K14-R125C-mediated filament disruption in keratinocytes and inhibited dispase-triggered keratinocyte sheet fragmentation and Fas-mediated apoptosis. Therefore, keratin acetylation may provide a novel therapeutic target for some keratin-associated diseases.

Water distribution in human hair microstructure elucidated by spin contrast variation small-angle neutron scattering

Dynamic nuclear polarization (DNP) is effective for controlling the neutron scattering length of protons and can be utilized for contrast variation in small-angle neutron scattering (SANS). Using the TEMPOL solution soaking method as electron spin doping, the DNP–SANS technique was applied to human hair fiber for the first time. For dry and D2O-swollen hair samples, a drastic change in the SANS profile was observed at high polarization conditions (|P H P N| ∼ 60%, where P H and P N are the proton and neutron spin polarization, respectively). The SANS profile as a function of the magnitude of the scattering vector, q, was composed of a low-q upturn, a middle-q oscillation and a high-q flat region. The low-q upturn was assumed to be a combination of two power-law functions, q −4 due to a large structure interface (Porod's law) and q −2 due to random coil. The middle-q oscillation was well reproduced by numerical calculation based on the structure model of intermediate filaments (IFs) as proposed by Er Rafik et al. [Biophys. J. (2004), 86, 3893–3904]: one pair of keratin coiled-coils is located at the center and surrounded by seven pairs of keratin coiled-coils located in a circle (called the `7 + 1' model), and a collection of IFs is arranged in a quasi-hexagonal manner. For the observed SANS profiles for different P H P N, the IF term contribution maintained a constant q-dependent profile, despite significant changes in intensity. This indicates that the macrofibril is composed of two domains (keratin coiled-coils and matrix). In addition, D2O swelling enhanced the IF term intensity and shifted the polarization-dependent local minimum to higher P H P N. This behavior was reproduced by contrast factor calculation based on the two-domain model. Scattering length densities of keratin coiled-coil and surrounding matrix domains were calculated by use of the known amino acid composition, considering the hydrogen–deuterium exchange reaction during soaking with D2O solution of TEMPOL. As a result, it was found that for keratin coiled-coil domains, about 40% of the peptide backbone amide NH protons were replaced with deuterons. This means that 68% of the α-helix domain is rigid, but the rest is flexible to allow dynamic dissociation of the hydrogen bond. Furthermore, the local mass density of each domain was precisely evaluated. The obtained data are expected to be a guide for further detailed investigation of keratin and keratin-associated protein distribution. This approach is expected to be applied to a wide variety of bio-derived materials, which are water absorbing in general.

An in vitro analysis of the hemostatic efficacy of fibrinogen precipitation with varied keratin fraction compositions

In preclinical studies, human hair has demonstrated effective hemostatic properties, potentially attributed to keratin proteins facilitating rapid conversion of fibrinogen to fibrin during coagulation. However, the rational use of human hair keratin for hemostasis remains unclear, given its complex mixture of proteins with diverse molecular weights and structures, leading to variable hemostatic capacity. To optimize the rational utilization of human hair keratin for hemostasis, we investigated the effects of different keratin fractions on keratin-mediated fibrinogen precipitation using a fibrin generation assay. Our study focused on high molecular weight keratin intermediate filaments (KIFs) and lower molecular weight keratin-associated proteins (KAPs) combined in various ratios during the fibrin generation. Scanning electron microscope analysis of the precipitates revealed a filamentous pattern with a broad distribution of fiber diameters, likely due to the diversity of keratin mixtures involved. An equal proportion of KIFs and KAPs in the mixture yielded the most extensive precipitation of soluble fibrinogen in an in vitro study, potentially due to structure-induced exposure of active sites. However, all hair protein samples exhibited diverse catalytic behaviors compared to thrombin, highlighting the potential of utilizing specific hair fractions to develop hair protein-based hemostatic materials with optimized capacity.

Current Progress and Biotechnological Applications of Microbial Keratinases

Keratin is a fibrous and recalcitrant protein found in feathers, nails, horns, hooves, and the epidermis of the skin. The presence of the high degree of disulfide bonds, hydrogen bonds, and hydrophobic interactions makes them resistant to mechanical stress and are not degraded by common proteases such as trypsin, pepsin, and papain. Due to the slow degradation of keratinous protein, accumulation of solid wastes from the poultry, slaughterhouse, textile, and leather industries leads to solid waste problems and other environmental and health related problems. In this review, efficient biodegradation of keratinous wastes by microorganisms, as a low-cost, environmentally friendly strategy has been discussed. Keratinases are the microbial proteases and hydrolyze the hard keratin. The decomposition of keratin by keratinases maintains the original structure of the final products, including short peptides, amino acids, and organic nitrogen which are deteriorated when traditional or chemical method is implemented. In this article, the role of keratinases producing bacterial and fungal species and their attributes has been elaborated, along with the biochemical characteristics of keratinases, and further, protein engineering approaches has been discussed, with the prospects to enhance keratinases activity for their biotechnological applications.