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1.
Curr Microbiol ; 81(8): 227, 2024 Jun 16.
Artigo em Inglês | MEDLINE | ID: mdl-38879855

RESUMO

Microbial degradation of keratin is characterized by its inherent safety, remarkable efficiency, and the production of copious degradation products. All these attributes contribute to the effective management of waste materials at high value-added and in a sustainable manner. Microbial degradation of keratin materials remains unclear, however, with variations observed in the degradation genes and pathways among different microorganisms. In this study, we sequenced the transcriptome of Purpureocillium lilacinum GZAC18-2JMP mycelia on control medium and the medium containing 1% feather powder, analyzed the differentially expressed genes, and revealed the degradation mechanism of chicken feathers by P. lilacinum GZAC18-2JMP. The results showed that the chicken feather degradation rate of P. lilacinum GZAC18-2JMP reached 64% after 216 h of incubation in the fermentation medium, reaching a peak value of 148.9 µg·mL-1 at 192 h, and the keratinase enzyme activity reached a peak value of 211 U·mL-1 at 168 h, which revealed that P. lilacinum GZAC18-2JMP had a better keratin degradation effect. A total of 1001 differentially expressed genes (DEGs) were identified from the transcriptome database, including 475 upregulated genes and 577 downregulated genes. Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis of the DEGs revealed that the metabolic pathways related to keratin degradation were mainly sulfur metabolism, ABC transporters, and amino acid metabolism. Therefore, the results of this study provide an opportunity to gain further insight into keratin degradation and promote the biotransformation of feather wastes.


Assuntos
Plumas , Hypocreales , Queratinas , Transcriptoma , Queratinas/metabolismo , Hypocreales/genética , Hypocreales/metabolismo , Animais , Plumas/metabolismo , Galinhas , Perfilação da Expressão Gênica , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Peptídeo Hidrolases/metabolismo , Peptídeo Hidrolases/genética , Micélio/genética , Micélio/metabolismo , Micélio/crescimento & desenvolvimento , Fermentação , Biodegradação Ambiental
2.
PLoS One ; 19(6): e0304495, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38875228

RESUMO

Discerning assimilated diets of wild animals using stable isotopes is well established where potential dietary items in food webs are isotopically distinct. With the advent of mixing models, and Bayesian extensions of such models (Bayesian Stable Isotope Mixing Models, BSIMMs), statistical techniques available for these efforts have been rapidly increasing. The accuracy with which BSIMMs quantify diet, however, depends on several factors including uncertainty in tissue discrimination factors (TDFs; Δ) and identification of appropriate error structures. Whereas performance of BSIMMs has mostly been evaluated with simulations, here we test the efficacy of BSIMMs by raising domestic broiler chicks (Gallus gallus domesticus) on four isotopically distinct diets under controlled environmental conditions, ideal for evaluating factors that affect TDFs and testing how BSIMMs allocate individual birds to diets that vary in isotopic similarity. For both liver and feather tissues, δ13C and δ 15N values differed among dietary groups. Δ13C of liver, but not feather, was negatively related to the rate at which individuals gained body mass. For Δ15N, we identified effects of dietary group, sex, and tissue type, as well as an interaction between sex and tissue type, with females having higher liver Δ15N relative to males. For both tissues, BSIMMs allocated most chicks to correct dietary groups, especially for models using combined TDFs rather than diet-specific TDFs, and those applying a multiplicative error structure. These findings provide new information on how biological processes affect TDFs and confirm that adequately accounting for variability in consumer isotopes is necessary to optimize performance of BSIMMs. Moreover, results demonstrate experimentally that these models reliably characterize consumed diets when appropriately parameterized.


Assuntos
Teorema de Bayes , Isótopos de Carbono , Galinhas , Isótopos de Nitrogênio , Animais , Galinhas/crescimento & desenvolvimento , Feminino , Isótopos de Carbono/análise , Masculino , Isótopos de Nitrogênio/análise , Dieta/veterinária , Fígado/metabolismo , Plumas/química , Plumas/metabolismo , Cadeia Alimentar , Modelos Biológicos
3.
Commun Biol ; 7(1): 592, 2024 May 17.
Artigo em Inglês | MEDLINE | ID: mdl-38760591

RESUMO

Limited studies using animal models with a few natural mutations in melanophilin (Mlph) provided partial functions of Mlph in melanosome trafficking. To investigate cellular functions of Mlph, especially ZnF motif of Mlph, we analyzed all three Mlph knockout (KO) quail lines, one and two base pair (bp) deletions as models for total KO, and three bp deletion causing deletion of one Cysteine (C84del) in the ZnF motif. All quail lines had diluted feather pigmentation with impaired dendritogenesis and melanosome transport in melanocytes. In vitro studies revealed capability of binding of the ZnF motif to PIP3, and impairment of PI3P binding and mislocalization of MLPH proteins with ZnF motif mutations. The shortened melanocyte dendrites by the C84del mutation were rescued by introducing WT Mlph in vitro. These results revealed the diluted feather pigmentation by Mlph mutations resulted from congregation of melanosomes in the cell bodies with impairment of the dendritogenesis and the transport of melanosomes to the cell periphery.


Assuntos
Plumas , Melanócitos , Melanossomas , Pigmentação , Animais , Plumas/metabolismo , Melanócitos/metabolismo , Pigmentação/genética , Melanossomas/metabolismo , Codorniz , Mutação , Proteínas Adaptadoras de Transdução de Sinal/genética , Proteínas Adaptadoras de Transdução de Sinal/metabolismo
4.
Nat Commun ; 15(1): 4174, 2024 May 16.
Artigo em Inglês | MEDLINE | ID: mdl-38755126

RESUMO

The transition from natal downs for heat conservation to juvenile feathers for simple flight is a remarkable environmental adaptation process in avian evolution. However, the underlying epigenetic mechanism for this primary feather transition is mostly unknown. Here we conducted time-ordered gene co-expression network construction, epigenetic analysis, and functional perturbations in developing feather follicles to elucidate four downy-juvenile feather transition events. We report that extracellular matrix reorganization leads to peripheral pulp formation, which mediates epithelial-mesenchymal interactions for branching morphogenesis. α-SMA (ACTA2) compartmentalizes dermal papilla stem cells for feather renewal cycling. LEF1 works as a key hub of Wnt signaling to build rachis and converts radial downy to bilateral symmetry. Novel usage of scale keratins strengthens feather sheath with SOX14 as the epigenetic regulator. We show that this primary feather transition is largely conserved in chicken (precocial) and zebra finch (altricial) and discuss the possibility that this evolutionary adaptation process started in feathered dinosaurs.


Assuntos
Galinhas , Plumas , Tentilhões , Animais , Plumas/crescimento & desenvolvimento , Plumas/metabolismo , Galinhas/genética , Tentilhões/genética , Regulação da Expressão Gênica no Desenvolvimento , Matriz Extracelular/metabolismo , Epigênese Genética , Redes Reguladoras de Genes , Via de Sinalização Wnt , Queratinas/metabolismo , Queratinas/genética , Evolução Biológica , Morfogênese/genética
5.
Curr Microbiol ; 81(7): 179, 2024 May 18.
Artigo em Inglês | MEDLINE | ID: mdl-38761211

RESUMO

Enormous aggregates of keratinous wastes are produced annually by the poultry and leather industries which cause environmental degradation globally. To combat this issue, microbially synthesized extracellular proteases known as keratinase are used widely which is effective in degrading keratin found in hair and feathers. In the present work, keratinolytic bacteria were isolated from poultry farm soil and feather waste, and various cultural conditions were optimized to provide the highest enzyme production for efficient keratin waste degradation. Based on the primary and secondary screening methods, the potent keratinolytic strain (HFS_F2T) with the highest enzyme activity 32.65 ± 0.16 U/mL was genotypically characterized by 16S rRNA sequencing and was confirmed as Bacillus velezensis HFS_F2T ON556508. Through one-variable-at-a-time approach (OVAT), the keratinase production medium was optimized with sucrose (carbon source), beef extract (nitrogen source) pH-7, inoculum size (5%), and incubation at 37 °C). The degree of degradation (%DD) of keratin wastes was evaluated after 35 days of degradation in the optimized keratinase production medium devoid of feather meal under submerged fermentation conditions. Further, the deteriorated keratin wastes were visually examined and the hydrolysed bovine hair with 77.32 ± 0.32% degradation was morphologically analysed through Field Emission Scanning Electron Microscopy (FESEM) to confirm the structural disintegration of the cuticle. Therefore, the current study would be a convincing strategy for reducing the detrimental impact of pollutants from the poultry and leather industries by efficient keratin waste degradation through the production of microbial keratinase.


Assuntos
Bacillus , Biodegradação Ambiental , Meios de Cultura , Plumas , Queratinas , Peptídeo Hidrolases , Bacillus/metabolismo , Bacillus/genética , Bacillus/enzimologia , Queratinas/metabolismo , Peptídeo Hidrolases/metabolismo , Peptídeo Hidrolases/genética , Animais , Plumas/metabolismo , Meios de Cultura/química , Aves Domésticas , RNA Ribossômico 16S/genética , Bovinos , Microbiologia do Solo , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/genética , Fermentação , Cabelo
6.
BMC Genomics ; 25(1): 505, 2024 May 22.
Artigo em Inglês | MEDLINE | ID: mdl-38778258

RESUMO

BACKGROUND: In day-old Hungarian white goose goslings, there is a noticeable difference in dorsal down coloration between males and females, with females having darker dorsal plumage and males having lighter plumage. The ability to autosex day-old goslings based on their dorsal down coloration is important for managing them efficiently and planning their nutrition in the poultry industry. The aim of this study was to determine the biological and genetic factors underlying this difference in dorsal down colorationthrough histological analysis, biochemical assays, transcriptomic profiling, and q‒PCR analysis. RESULTS: Tissue analysis and biochemical assays revealed that compared with males, 17-day-old embryos and day-old goslings of female geese exhibited a greater density of melanin-containing feather follicles and a greater melanin concentration in these follicles during development. Both female and male goslings had lower melanin concentrations in their dorsal skin compared to 17-day-old embryos. Transcriptome analysis identified a set of differentially expressed genes (DEGs) (MC1R, TYR, TYRP1, DCT and MITF) associated with melanogenesis pathways that were downregulated or silenced specifically in the dorsal skin of day-old goslings compared to 17-day-old embryos, affecting melanin synthesis in feather follicles. Additionally, two key genes (MC1R and MITF) associated with feather coloration showed differences between males and females, with females having higher expression levels correlated with increased melanin synthesis and darker plumage. CONCLUSION: The expression of multiple melanogenesis genes determines melanin synthesis in goose feather follicles. The dorsal down coloration of day-old Hungarian white goose goslings shows sexual dimorphism, likely due to differences in the expression of the MC1R and MITF genes between males and females. These results could help us better understand why male and female goslings exhibit different plumage patterns.


Assuntos
Gansos , Perfilação da Expressão Gênica , Melaninas , Pigmentação , Caracteres Sexuais , Animais , Feminino , Masculino , Gansos/genética , Gansos/metabolismo , Melaninas/metabolismo , Pigmentação/genética , Plumas/metabolismo , Plumas/crescimento & desenvolvimento , Transcriptoma
7.
PLoS Biol ; 22(5): e3002636, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38743770

RESUMO

Periodic patterning requires coordinated cell-cell interactions at the tissue level. Turing showed, using mathematical modeling, how spatial patterns could arise from the reactions of a diffusive activator-inhibitor pair in an initially homogeneous 2D field. Most activators and inhibitors studied in biological systems are proteins, and the roles of cell-cell interaction, ions, bioelectricity, etc. are only now being identified. Gap junctions (GJs) mediate direct exchanges of ions or small molecules between cells, enabling rapid long-distance communications in a cell collective. They are therefore good candidates for propagating nonprotein-based patterning signals that may act according to the Turing principles. Here, we explore the possible roles of GJs in Turing-type patterning using feather pattern formation as a model. We found 7 of the 12 investigated GJ isoforms are highly dynamically expressed in the developing chicken skin. In ovo functional perturbations of the GJ isoform, connexin 30, by siRNA and the dominant-negative mutant applied before placode development led to disrupted primary feather bud formation. Interestingly, inhibition of gap junctional intercellular communication (GJIC) in the ex vivo skin explant culture allowed the sequential emergence of new feather buds at specific spatial locations relative to the existing primary buds. The results suggest that GJIC may facilitate the propagation of long-distance inhibitory signals. Thus, inhibition of GJs may stimulate Turing-type periodic feather pattern formation during chick skin development, and the removal of GJ activity would enable the emergence of new feather buds if the local environment were competent and the threshold to form buds was reached. We further propose Turing-based computational simulations that can predict the sequential appearance of these ectopic buds. Our models demonstrate how a Turing activator-inhibitor system can continue to generate patterns in the competent morphogenetic field when the level of intercellular communication at the tissue scale is modulated.


Assuntos
Comunicação Celular , Plumas , Junções Comunicantes , Animais , Junções Comunicantes/metabolismo , Plumas/crescimento & desenvolvimento , Plumas/metabolismo , Embrião de Galinha , Conexinas/metabolismo , Conexinas/genética , Padronização Corporal/fisiologia , Galinhas , Pele/metabolismo , Isoformas de Proteínas/metabolismo , Isoformas de Proteínas/genética
8.
Dev Biol ; 508: 64-76, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38190932

RESUMO

Feathers originate as protofeathers before birds, in pterosaurs and basal dinosaurs. What characterizes a feather is not only its outgrowth, but its barb cells differentiation and a set of beta-corneous proteins. Reticula appear concomitantly with feathers, as small bumps on plantar skin, made only of keratins. Avian scales, with their own set of beta-corneous proteins, appear more recently than feathers on the shank, and only in some species. In the chick embryo, when feather placodes form, all the non-feather areas of the integument are already specified. Among them, midventral apterium, cornea, reticula, and scale morphogenesis appear to be driven by negative regulatory mechanisms, which modulate the inherited capacity of the avian ectoderm to form feathers. Successive dermal/epidermal interactions, initiated by the Wnt/ß-catenin pathway, and involving principally Eda/Edar, BMP, FGF20 and Shh signaling, are responsible for the formation not only of feather, but also of scale placodes and reticula, with notable differences in the level of Shh, and probably FGF20 expressions. This sequence is a dynamic and labile process, the turning point being the FGF20 expression by the placode. This epidermal signal endows its associated dermis with the memory to aggregate and to stimulate the morphogenesis that follows, involving even a re-initiation of the placode.


Assuntos
Ectoderma , Plumas , Animais , Embrião de Galinha , Plumas/metabolismo , Ectoderma/metabolismo , Evolução Biológica , Aves , Queratinas/metabolismo , Morfogênese
9.
Int J Biol Macromol ; 260(Pt 2): 129659, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38266845

RESUMO

KerJY-23 was a novel keratinase from feather-degrading Ectobacillus sp. JY-23, but its enzymatic characterization and structure are still unclear. In this study, the KerJY-23 was obtained by heterologous expression in Escherichia coli BL21(DE3), and enzymatic properties indicated that KerJY-23 was optimal at 60 °C and pH 9.0 and could be promoted by divalent metal ions or reducing agents. Furthermore, KerJY-23 had a broad substrate specificity towards casein, soluble keratin, and expanded feather powder, but its in vitro degradation against chicken feathers required an additional reducing agent. Homology modeling indicated that KerJY-23 contained a highly conserved zinc-binding HELTH motif and a His-Asp-Ser catalytic triad that belonged to the typical characteristics of M4-family metallo-keratinase and serine-keratinase, respectively. Molecular docking revealed that KerJY-23 achieved a reinforced binding on feather keratin via abundant hydrogen bonding interactions. This work not only deepened understanding of the novel and interesting metallo-serine keratinase KerJY-23, but also provided a theoretical basis for realizing the efficient use of waste feather keratin.


Assuntos
Galinhas , Serina , Animais , Serina/metabolismo , Galinhas/metabolismo , Simulação de Acoplamento Molecular , Peptídeo Hidrolases/metabolismo , Plumas/metabolismo , Queratinas/metabolismo , Concentração de Íons de Hidrogênio , Temperatura
10.
J Sci Food Agric ; 104(9): 5176-5185, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38284560

RESUMO

BACKGROUND: The present study was conducted to investigate the effects of dietary novel alkaline protease from Bacillus licheniformis on the growth performance, meat quality, antioxidant status and intestinal morphology of broilers. In total, 4000 broilers were randomly assigned into five groups and treated with normal control, normal control + 100 mg kg-1 protease, normal control + 200 mg kg-1 protease, normal control + 300 mg kg-1 protease and normal control + 400 mg kg-1 protease. RESULTS: Supplementing protease impacted final body weight (linear, P = 0.003; quadratic, P = 0.006) and decreased feed conversion rate (linear, P = 0.036) in broilers. Moreover, dietary protease significantly increased breast muscle rate (linear, P = 0.005; quadratic, P = 0.021) and decreased drip loss (linear, P < 0.001; quadratic, P < 0.001). In addition, dietary protease notably increased protein digestibility (linear, P = 0.001; quadratic, P = 0.006) and trypsin activity (linear, P = 0.002; quadratic, P = 0.009) in jejunum. Light microscopy revealed that the jejunum villi in the 300 mg kg-1 and 400 mg kg-1 groups exhibited greater height and a denser arrangement compared to those in the control group. The addition of protease decreased malondialdehyde content (linear, P < 0.001; quadratic, P < 0.001) and increased total antioxidant capacity (linear, P = 0.001; quadratic, P < 0.001) in pectoral muscles. CONCLUSION: The results of the present study suggest that dietary novel alkaline protease from B. licheniformis improved growth performance by affecting trypsin activity, protein digestibility, antioxidant capacity and intestinal health. © 2024 Society of Chemical Industry.


Assuntos
Ração Animal , Antioxidantes , Bacillus licheniformis , Proteínas de Bactérias , Galinhas , Endopeptidases , Intestinos , Carne , Animais , Galinhas/crescimento & desenvolvimento , Galinhas/metabolismo , Bacillus licheniformis/enzimologia , Bacillus licheniformis/crescimento & desenvolvimento , Bacillus licheniformis/metabolismo , Antioxidantes/metabolismo , Endopeptidases/metabolismo , Endopeptidases/química , Ração Animal/análise , Carne/análise , Intestinos/crescimento & desenvolvimento , Proteínas de Bactérias/metabolismo , Masculino , Suplementos Nutricionais/análise , Plumas/química , Plumas/metabolismo , Plumas/crescimento & desenvolvimento , Dieta/veterinária , Digestão
11.
Bioresour Technol ; 393: 130110, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38040301

RESUMO

Feather waste, a by-product of the poultry industry, is rich in proteins, peptides, and amino acids. Improper disposal of feathers can cause environmental pollution. Solid-state fermentation (SSF) is a viable alternative to submerged fermentation due to its simplicity, productivity, and lower cost. The study goal is a biorefinery of chicken feather waste supplemented with wheat bran using a recombinant Bacillus subtilis strain to produce soluble proteins and a serine alkaline protease. Plackett-Burman Design and Central Composite Design were utilized in a statistical-mathematical model to optimize the process. Multi-factorial design optimization resulted in 80 % substrate degradation efficiency, an alkaline protease with dual activities (1423 proteolytic units and 190 keratinolytic units), 214 mg soluble proteins/g substrate, and 87 % model validation. Scaling up the SSF process to 50 g of substrate significantly enhanced the end products of feather biodegradation to 1616 proteolytic units, 2844 keratinolytic units, and 127 mg soluble proteins/g substrate. AIM AND SCOPE OF THE MANUSCRIPT: The aim of the present study is to utilize chicken feather waste (alone or supplemented with other materials) through recombinant Bacillus subtilis cells using solid state fermentation (SSF) at a laboratory scale. The plan study provides a promising waste management in the environmental field concerning biodegradation of such recalcitrant keratinous wastes supplemented with agricultural residues via recombinant microorganism. On semi-pilot scale, high production and quality of soluble protein, protease, and keratinase activity were produced according to the statistically optimised first stage fermentation in the laboratory scale. The bioconversion process took place as a major goal to obtain valuable products, with low utilities and energy requirements. Therefore, this will consider as an economically feasible and environmentally friendly alternative. Moreover, this study is considered as first step fermentation for feather waste to pave the road for directing it to a second step fermentation for biogas production and bioenergy generation through bio-electrochemical systems (Manuscript under publication).


Assuntos
Bacillus subtilis , Proteínas de Bactérias , Galinhas , Endopeptidases , Animais , Bacillus subtilis/metabolismo , Fermentação , Galinhas/metabolismo , Serina/análise , Serina/metabolismo , Plumas/química , Plumas/metabolismo , Peptídeo Hidrolases/metabolismo
12.
G3 (Bethesda) ; 14(2)2024 Feb 07.
Artigo em Inglês | MEDLINE | ID: mdl-37943814

RESUMO

Bird plumage coloration is a complex and multifactorial process that involves both genetic and environmental factors. Diverse pigment groups contribute to plumage variation in different birds. In parrots, the predominant green color results from the combination of 2 different primary colors: yellow and blue. Psittacofulvin, a pigment uniquely found in parrots, is responsible for the yellow coloration, while blue is suggested to be the result of light scattering by feather nanostructures and melanin granules. So far, genetic control of melanin-mediated blue coloration has been elusive. In this study, we demonstrated that feather from the yellow mutant rose-ringed parakeet displays loss of melanosome granules in spongy layer of feather barb. Using whole genome sequencing, we found that mutation in SLC45A2, an important solute carrier protein in melanin synthetic pathway, is responsible for the sex-linked yellow phenotype in rose-ringed parakeet. Intriguingly, one of the mutations, P53L found in yellow Psittacula krameri is already reported as P58A/S in the human albinism database, known to be associated with human OCA4. We further showed that mutations in SLC45A2 gene affect melanin production also in other members of Psittaculidae family such as alexandrine and plum-headed parakeets. Additionally, we demonstrate that the mutations associated with the sex-linked yellow phenotype, localized within the transmembrane domains of the SLC45A2 protein, affect the protein localization pattern. This is the first evidence of plumage color variation involving SLC45A2 in parrots and confirmation of associated mutations in the transmembrane domains of the protein that affects its localization.


Assuntos
Melaninas , Papagaios , Humanos , Animais , Melaninas/genética , Plumas/química , Plumas/metabolismo , Mutação , Papagaios/metabolismo , Fenótipo , Pigmentação/genética , Antígenos de Neoplasias/genética , Antígenos de Neoplasias/metabolismo , Proteínas de Membrana Transportadoras/genética
13.
Gen Comp Endocrinol ; 345: 114390, 2024 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-37844650

RESUMO

Wild animals have been increasingly exposed to a wide range of stressors, mainly due to the intensification of human activities and habitat modifications. Consequently, new tools in order to assess the physiological and health status of wild animals have been developed. In particular, glucocorticoids have received a special attention. Primarily metabolic hormones, they are also used to evaluate the stress level of organisms. While historically measured in blood samples, new less-invasive methods have been recently developed to measure glucocorticoids in matrices such as faeces, hairs/feathers, or saliva. To date, measurements in saliva are still in their infancy despite the numerous advantages of the matrix: non-invasive, reflects the biologically active portion of glucocorticoids, allows to measure both baseline and stress-induced levels. In addition, most studies using saliva have been performed on domestic and captive animals, and recent development in wild animals have focused on mammals. Here, we show, for the first time, that saliva could also be reliably used in free-ranging birds, as glucocorticoid levels in saliva strongly correlated with plasma levels. This promising result opens new avenues for a non-invasive sampling method to assess health status of wild birds in conservation biology and ecology.


Assuntos
Corticosterona , Glucocorticoides , Animais , Humanos , Glucocorticoides/metabolismo , Animais Selvagens/metabolismo , Aves/metabolismo , Plumas/metabolismo , Mamíferos/metabolismo
14.
Sci Rep ; 13(1): 19989, 2023 11 15.
Artigo em Inglês | MEDLINE | ID: mdl-37968282

RESUMO

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.


Assuntos
Escherichia coli , Plumas , Animais , Plumas/metabolismo , Escherichia coli/genética , Simulação de Acoplamento Molecular , Pichia/metabolismo , Metaloproteases/metabolismo , Queratinas/genética , Queratinas/metabolismo , Clonagem Molecular
15.
Int J Biol Macromol ; 253(Pt 6): 127317, 2023 Dec 31.
Artigo em Inglês | MEDLINE | ID: mdl-37820911

RESUMO

Purified calcium serine metalloprotease from Stenotrophomonas maltophilia strain SMPB12 exhibits highest enzyme activity at pH 9 and temperature range between 15 °C-25 °C. Enzyme supplemented with 40 µM Ca-Hap-NP (NP-protease) showed maximum elevated activity of 17.29 µmole/min/ml (1.9-fold of original protease activity). The thermostability of the enzyme was maintained for 1 h at 60 °C over an alkaline pH range 7.5-10, as compared to the NP untreated enzyme whose activity was of 8.97 µmole/min/ml. A significant loss of activity with EDTA (1.05 µmole/min/ml, 11.75 %), PMSF (0.93 µmole/min/ml, 10.46 %) and Hg2+ (3.81 µmole/min/ml, 42.49 %) was also observed. Kinetics study of NP-protease showed maximum decreases in Km (28.11 %) from 0.28 mM (NP untreated enzyme) to 0.22 mM (NP-protease) along with maximum increase in Vmax (42.88 %) from 1.25 µmole/min/ml to 1.79 µmole/min/ml at varying temperatures. The enhanced activity of NP-protease was able to efficiently degrade recalcitrant solid wastes like feather to produce value-added products like amino acids and helps in declogging recalcitrant solid wastes. The nano-enabled protease may be utilized in a smaller amount for degrading in bulk recalcitrant solid proteinaceous waste at 15 °C temperature as declogging agents providing an eco-friendly efficient process.


Assuntos
Durapatita , Plumas , Animais , Plumas/metabolismo , Durapatita/metabolismo , Resíduos Sólidos , Peptídeo Hidrolases/metabolismo , Bactérias/metabolismo , Temperatura , Florestas , Concentração de Íons de Hidrogênio , Estabilidade Enzimática
16.
Appl Microbiol Biotechnol ; 107(23): 7055-7070, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37750916

RESUMO

Feathers become hazardous pollutants when deposited directly into the environment. The rapid expansion of the poultry industry has significantly increased feather waste, necessitating the development of new ways to degrade and utilize feathers. This study investigated the ability of Bacillus licheniformis WHU to digest intact chicken feathers in water. The results indicated that yields of free amino acids, bioactive peptides, and keratin-derived nano-/micro-particles were improved in bacteria- versus purified keratinase-derived feather hydrolysate. Bacteria-derived feather hydrolysate supplementation induced health benefits in mice, including significantly increased intestinal villus height and zonula occludens-1 protein expression, as well as increased secretory immunoglobulin A levels in the intestinal mucosa and superoxide dismutase activity in serum. Additionally, feather hydrolysate supplementation modulated the mouse gut microbiota, reflected by increased relative abundance of probiotics such as Lactobacillus spp., decreased relative abundance of Proteobacteria at the phylum level and pathogens such as Staphylococcus spp., and increased Bacteroidota/Firmicutes ratio. This study developed a simple, cost-effective method to degrade feathers by B. licheniformis WHU digestion, yielding a hydrolysate that can be directly used as a bioactive nutrient resource. The study findings have applications in the livestock, poultry, and aquaculture industries, which have high demands for cheap protein. KEY POINTS: • Bacillus licheniformis could degrade intact feather in water. • The resulting feather hydrolysate shows prebiotic effects on mouse.


Assuntos
Bacillus licheniformis , Animais , Camundongos , Bacillus licheniformis/metabolismo , Plumas/química , Plumas/metabolismo , Plumas/microbiologia , Água/metabolismo , Galinhas , Peptídeo Hidrolases/metabolismo , Aves Domésticas , Bactérias/metabolismo , Nutrientes , Queratinas/metabolismo
17.
Artigo em Inglês | MEDLINE | ID: mdl-37390888

RESUMO

Measuring corticosterone in feathers allows researchers to make long-term, retrospective assessments of physiology with non-invasive sampling. To date, there is little evidence that steroids degrade within the feather matrix, however this has yet to be determined from the same sample over many years. In 2009, we made a pool of European starling (Sturnus vulgaris) feathers that had been ground to a homogenous powder using a ball mill and stored on a laboratory bench. Over the past 14 years, a subset of this pooled sample has been assayed via radioimmunoassay (RIA) 19 times to quantify corticosterone. Despite high variability across time (though low variability within assays), there was no effect of time on measured feather corticosterone concentration. In contrast, two enzyme immunoassays (EIA) produced higher concentrations than the samples assayed with RIA, though this difference is likely due to different binding affinities of the antibodies used. The present study provides further support for researchers to use specimens stored long-term and from museums for feather corticosterone quantification, and likely applies to corticosteroid measurements in other keratinized tissues.


Assuntos
Corticosterona , Plumas , Animais , Corticosterona/metabolismo , Plumas/metabolismo , Estudos Retrospectivos
18.
Anim Biotechnol ; 34(9): 5001-5010, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37300547

RESUMO

To explore the relationship between PMEL gene and quail plumage color, to provide a reference for subsequent quail plumage color breeding. In this experiment, RT-qPCR technology was used to analyze the relative mRNA expression levels of Korean quail (maroon) and Beijing white quail embryos at different developmental stages. Two SNPs in PMEL gene were screened based on the RNA-Seq data of skin tissues of Korean quail and Beijing white quail during embryonic stage. The KASP technology was used for genotyping in the resource population and correlation analysis was carried out with the plumage color traits of quail. Finally, the bioinformatics technology was used to predict the effects of these two SNPs on the structure and function of the encoded protein. The results showed that the expression levels of PMEL gene during the embryonic development of Beijing white quail were extremely significantly higher than that of Korean quail (p < 0.01). The frequency distribution of the three genotypes (AA, AB, and BB) of the Beijing white quail at the c. 1030C > T and c. 1374A > G mutation sites were extremely significantly different from that of the Korean quail (p < 0.01). And there was a significant correlation between the c. 1374A > G mutation site with white plumage phenotype. Bioinformatics analysis showed that SNP1 (c. c1030t) located in exon 6 was a harmful mutation site, and SNP2 (c. a1374g) located in exon 7 was a neutral mutation site. Protein conservation prediction showed that the coding protein P344S site caused by SNP1 (c. c1030t) site and the coding protein I458M site caused by SNP2 (c. g2129a) site were non-conservative sites. The results of this experiment showed that the PMEL gene was associated with the plumage color traits of quail and could be used as a candidate gene for studying the plumage color of quail.


Assuntos
Polimorfismo de Nucleotídeo Único , Codorniz , Animais , Polimorfismo de Nucleotídeo Único/genética , Codorniz/genética , Plumas/metabolismo , Coturnix/genética , Pigmentação/genética , Expressão Gênica
19.
Genes (Basel) ; 14(6)2023 05 29.
Artigo em Inglês | MEDLINE | ID: mdl-37372372

RESUMO

Leveraging computation in the development of peptide therapeutics has garnered increasing recognition as a valuable tool to generate novel therapeutics for disease-related targets. To this end, computation has transformed the field of peptide design through identifying novel therapeutics that exhibit enhanced pharmacokinetic properties and reduced toxicity. The process of in-silico peptide design involves the application of molecular docking, molecular dynamics simulations, and machine learning algorithms. Three primary approaches for peptide therapeutic design including structural-based, protein mimicry, and short motif design have been predominantly adopted. Despite the ongoing progress made in this field, there are still significant challenges pertaining to peptide design including: enhancing the accuracy of computational methods; improving the success rate of preclinical and clinical trials; and developing better strategies to predict pharmacokinetics and toxicity. In this review, we discuss past and present research pertaining to the design and development of in-silico peptide therapeutics in addition to highlighting the potential of computation and artificial intelligence in the future of disease therapeutics.


Assuntos
Inteligência Artificial , Plumas , Animais , Simulação de Acoplamento Molecular , Plumas/metabolismo , Peptídeos/farmacologia , Peptídeos/uso terapêutico , Peptídeos/química , Proteínas/metabolismo
20.
Microbiol Res ; 274: 127439, 2023 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-37364416

RESUMO

Microbial keratinases have prominent potential in biotransformation of recalcitrant keratin substrates to value-added products which has made keratinases a research focus in the past decades. In this study, an efficient feather-degrading bacterium was isolated and identified as a novel species in Ectobacillus genus and designated as Ectobacillus sp. JY-23. The degradation characteristics analysis revealed that Ectobacillus sp. JY-23 could utilize chicken feathers (0.4% w/v) as the sole nutrient source and degraded 92.95% of feathers in 72 h. A significant increase in sulfite and free sulfydryl group content detected in the feather hydrolysate (culture supernatant) indicated efficient reduction of disulfide bonds, which inferred that the degradation mechanism of isolated strain was a synergetic action of sulfitolysis and proteolysis. Moreover, abundant amino acids were also detected, among which proline and glycine were the predominant free amino acids. Then, the keratinase of Ectobacillus sp. JY-23 was mined and Y1_15990 was identified as the keratinase encoding gene of Ectobacillus sp. JY-23 and designated as kerJY-23. Escherichia coli strain overexpressing kerJY-23 degraded chicken feathers in 48 h. Finally, bioinformatics prediction of KerJY-23 demonstrated that it belonged to the M4 metalloprotease family, which was a third keratinase member in this family. KerJY-23 showed low sequence identity to the other two keratinase members, indicating the novelty of KerJY-23. Overall, this study presents a novel feather-degrading bacterium and a new keratinase in the M4 metalloprotease family with remarkable potential in feather keratin valorization.


Assuntos
Galinhas , Plumas , Animais , Plumas/metabolismo , Plumas/microbiologia , Peptídeo Hidrolases/metabolismo , Metaloproteases/metabolismo , Queratinas/metabolismo , Aminoácidos/metabolismo , Concentração de Íons de Hidrogênio
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