Pernyi Silkworm Research Articles

Silk as templates for hydroxyapatite biomineralization: A comparative study of Bombyx mori and Antheraea pernyi silkworm silks

Silk is extensively investigated in bone tissue engineering due to its extraordinary mechanical properties and ability to regulate biomineralization. Protein templates regulate biomineralization process through chemical interaction with ions. However, the effect of structural differences in silk fibroin on biomineralization has not been studied in detail. In this study, Antheraea pernyi silk fibroin (ASF) and Bombyx mori silk fibroin (BSF) fibers were used as templates to study the effect of silk species on biomineralization. The results showed that silk fibroin could induce the formation of calcium-deficient hydroxyapatite in simulated body fluid (SBF), and the SBF treatment resulted in the formation of silk I crystals. Compared with BSF, ASF exhibited a higher ability to induce mineralization, which may depend on the differences in hydrophilic amorphous fractions between ASF and BSF. The amorphous fractions of ASF contain more acidic amino acids, which can provide more nucleation sites in the initial stage of mineralization, resulting in faster mineralization process and more mineral deposits. This study decodes the key role of silk fibroin fractions on biomineralization, and provides deeper insights for the study of silk fibroin as biomineralization template and bone repair materials.

Understanding the Mechanical Properties and Structure Transition of Antheraea pernyi Silk Fiber Induced by Its Contraction.

Like most major ampullate silks of spider, the length of Antheraea pernyi silkworm silk can shrink to a certain degree when the fiber is in contact with water. However, what happens in terms of molecule chain level and how it correlates to the mechanical properties of the silk during its contraction is not yet fully understood. Here, we investigate the water-induced mechanical property changes as well as the structure transition of two kinds of A. pernyi silk fiber, which are forcibly reeled from two different individuals (silkworm a and silkworm b; the silk fiber from either one represents the lower and upper limit of the distribution of mechanical properties, respectively). The tensile test results present that most of the mechanical parameters except the post-yield modulus and breaking strain for both silk fibers have the same variation trend before and after their water contraction. Synchrotron FTIR and Raman spectra show that the native filament from silkworm a contains more α-helix structures than that in silkworm b filament, and these α-helices are partially converted to β-sheet structures after the contraction of the fibers, while the order of both β-sheet and α-helix slightly increase. On the other side, the content and orientation of both secondary structural components in silkworm b fiber keep unchanged, no matter if it is native or contracted. 13C CP/MAS NMR results further indicate that the α-helix/random coil to β-sheet conformational transition that occurred in the silk of silkworm a corresponds the Ala residues. Based upon these results, the detailed structure transition models of both as-reeled A. pernyi silk fibers during water contraction are proposed finally to interpret their properties transformation.

Comparative Study of Strain-Dependent Structural Changes of Silkworm Silks: Insight into the Structural Origin of Strain-Stiffening.

Structure-property relationships of silk is an intriguing topic for silk-based biomaterials research since these features are related to biomimicking the processing in natural silk fiber formation which results in excellent mechanical properties. Strain-stiffening is common for spider silks and nonmulberry silkworm silks. However, the structural origin of strain-stiffening remains unclear. In this paper, the strain-dependent structural change of Antheraea pernyi silkworm silk is studied by X-ray fiber diffraction and Fourier transform infrared spectroscopy under stretching. Based on a combination of mechanical and structural analysis, the molecular origins of strain-stiffening in A. pernyi silk were determined. The relatively high content of the β-sheets within the amorphous domains in A. pernyi silk is responsible for strain-stiffening, where "molecular spindles" enhance the extensibility and toughness of the fiber.

Precise correlation of macroscopic mechanical properties and microscopic structures of animal silks-using Antheraea pernyi silkworm silk as an example.

Animal silks, as one type of high performance natural material, display a unique combination of modulus, tensile strength, and extensibility that gives rise to a greater toughness than any other natural or synthetic fibers. Many previous researchers have already suggested that such excellent comprehensive mechanical properties should be closely related to their special molecular structures. In this paper, we provide more direct evidence to such an assumption by using Antheraea pernyi silkworm silk (tussah silk) as an example with synchrotron radiation FTIR microspectroscopy as a major characterization tool. Being a silkworm silk, A. pernyi silk has the same function as other silkworm silks (like common Bombyx mori silk), but on the other hand, its amino acid residue sequence is similar to that of spider dragline silk. Thus, A. pernyi silk can be a bridge between silkworm silk and spider silk that is worth investigating. Hence, in this research we designed different forcibly reeled A. pernyi silk samples by controlling the reeling rate, and subsequently tested their mechanical properties and then correlated them with their molecular structures and orientation degrees. Results show that the Young's modulus and breaking stress of forcibly reeled A. pernyi silks increased with the reeling rate, whereas the breaking strain was reduced. In the meantime, structure characterization revealed that the β-sheet content and molecular chain orientation in A. pernyi silk all increased significantly with an increase in reeling rate. In addition, the mechanical performance of A. pernyi silk can be altered from close to that of spider dragline silk to that of B. mori silkworm silk, with just a change of the reeling rate. All these phenomena clearly indicate that structural changes in A. pernyi silks contrived and controlled by reeling rate have a great effect upon their final mechanical properties. These observations further confirm that the mechanical properties of animal silks are able to be tuned by structure control during harvest time. Furthermore, the results obtained in this study may provide useful guidance when designing and producing high performance regenerated silk fibers for different applications.

Directional moisture transfer through a wild silkworm cocoon wall.

A silkworm cocoon is a porous biological structure with multiple protective functions. In the current work, the authors have used both experimental and numerical methods to reveal the unique moisture transfer characteristics through a wild Antheraea pernyi silkworm cocoon wall, in comparison with the long-domesticated Bombyx mori silkworm cocoon walls. The water vapor transmission and water vapor permeability (WVP) properties show that the A. pernyi cocoons exhibit directional moisture transfer behavior, with easier moisture transfer from inside out than outside in [e.g., the average WVP is 0.057 g/(h m bar) from inside out and is 0.034 g/(h m bar) from outside in]. Numerical analysis shows that the cubic mineral crystals in the outer section of the A. pernyi cocoon wall create a rough surface that facilitates air turbulence and promotes disturbance amplitude of the flow field, leading to lengthened water vapor transfer path and increased tortuosity of the moist air. It also indicates the vortex of water vapor can be generated in the outer section of cocoon wall, which increases the diffusion distance of water vapor and enhances the turbulence kinetic energy and turbulence eddy dissipation, signifying higher moisture resistance in the outer section. The difference in moisture resistance of the multiple A. pernyi cocoon layers is largely responsible for the unique directional moisture transfer behavior of this wild silkworm cocoon. These findings may inspire a biomimicry approach to develop novel lightweight moisture management materials and structures.

Interfacial heat transfer through a natural protective fibrous architecture: a wild silkworm cocoon wall

The wild Antheraea pernyi silkworm cocoon is a thin and light-weight structure, yet it has shown effective thermal insulation characteristics against extreme temperature fluctuations, which meet the demands of humans for lighter materials with higher thermal resistance. We present a two-dimensional computational fluid dynamics model of this unique fibrous cocoon structure to simulate the heat transfer process through the cocoon wall. The model is able to predict the temperature field inside the cocoon reasonably well. The results of the model also show that the mineral crystals present in the outer layers of the Antheraea pernyi cocoon can increase air flow resistance and decrease the effect of natural convection, which further reduces the heat transfer through the cocoon wall effectively. This has practical significance for the development of thermal functional textiles and composite structures.

Microstructure and mechanical properties of silk from different components of the Antheraea pernyi cocoon

Silk fibres from different components of the Antheraea pernyi silkworm cocoon, namely peduncle, outer floss, and cocoon shells (outermost layer and pelade layer) were studied in detail to gain insights into the structure–property–function relationship. Among the fibres from different components, peduncle fibres are the softest with the largest viscoelastic lag, which may reduce the oscillation amplitude when a cocoon hangs on a twig. Fibres from the outermost layer are the toughest and have the largest breaking energy. Outer floss fibres have the highest content of sericin (about 11.98%) but their hardness and elasticity are intermediate. Pelade fibres are shape – preservable and stable with superior hardness and elasticity. The understanding of the properties of different silk fibres is essential for understanding their respective roles in the function of a silk cocoon and will also inspire new designs of protective materials under stringent environmental conditions.

Calcium Nitrate/에탄올 혼합 용액에 의한 작잠견사(繭絲)의 용해

작잠은 가잠과는 종속이 다른 섬유 곤충으로 우수한 섬유용 재료를 만든다. 최근 비섬유용 소재로 작잠 견단백질을 활용하기 위한 기초 연구의 일환으로 작잠 견단백질을 용해 조건에 대하여 연구 결과 작잠견사의 연감율은 16.8%로 나타났으며, 작잠견사는 염화칼슘 에탄올 혼합 용매에는 용해되지 않으나 질산칼슘수화물의 용융액에는 용해되었다. 질산칼슘과 에탄올의 조성비를 조정하여 낮은 농도의 질산칼슘 농도에서도 작잠견사가 용해되었다. 작잠 실크단백질의 주요 아미노산 조성은 알라닌, 글리신, 세리신으로 작잠 정련사와 유사한 조성을 보였다. 따라서, 금후 작잠 견단백질의 적절한 용해시스템 확보 및 소재개발을 위한 기초 자료로 활용될 수 있을 것으로 사료된다. Antheraea pernyi silkworm is one of typical wild silkworms, which spins a tawny color cocoon. The cocoon has been used as a resource for textile material due to strong chemical stability and good mechanical properties. In this study, to increase the solubility efficiency of A. pernyi silk fibroin, the composition of dissolution solvent were examined. Calcium chloride tertiary system, normally used for dissolution of Bombyx mori silk fibroin, does not act on A. pernyi silk fibroin. Calcium nitrate system dissolves A. pernyi silk fibroin, and calcium nitrate ethanol system do more easily than calcium nitrate system. Amino acid composition of A. pernyi silk fibroin obtained after dissolution is mainly composed of alanine, glycine, and serine. A. pernyi silk fibroin would be used for non-textile applications near future.

Cocoon Characteristics of Antheraea pernyi Silkworm Reared in Korean Oak Field

Antheraea pernyi silkworm is a well known wild silkworm to produce a valuable silk fiber. A. pernyi silkworm was reared in Korean oak field and examined the cocoon characteristics, such as cocoon weight, cocoon shell weight, and percentage of cocoon shell weight. Degumming loss was also measured after alkali degumming process. A. pernyi silkworm spins tawny color cocoon in oval shape. Cocoon shell weight of A. pernyi silkworm, 0.78 g, was heavier than that of B. mori silkworm, 0.51 to 0.63 g. Cocoon shell percentage of A. pernyi silkworm, 32.8%, was higher than that of B. mori silkworm, 23.4 to 25.2%. Degumming loss percentage of A. pernyi silkworm, 17.1%, was lower than that of B. mori silkworm, 25.0%. SEM showed that the surface of the cocoon filament was coarse and oriented with longitudinal direction.

Structural and physical properties ofAntheraea pernyi silk fibroin fiber treated with I2-KI aqueous solution

Silk fibroin (SF) fiber from theAntheraea pernyi silkworm was treated with a 1.23 N iodine-potassium iodide (I2-KI) aqueous solution, and the structure and physical properties were investigated to clarify the effects of the iodine treatment. The noticeably high weight gain value of SF fiber, about 25 wt% was attributed to the absorption of polyiodide ions in the form of I3− and I5−. Fourier transform infrared spectroscopy and X-ray diffraction measurements suggested that polyiodide ions mainly entered the amorphous region. In addition, a new sharp reflection on the meridional direction, corresponding to a period of 7.0 A, was observed and indicated the possibility of the formation of mesophase structure ofβ-conformation chains. Dynamic viscoelastic measurements showed that the damping tanδ peak at 270°C gradually shifted to lower temperature in the iodinated SF fibers, suggesting an enhancement of the molecular motion of the fibroin chains induced by the presence of polyiodide ions. With heating above 254 °C, the iodine component introduced intermolecular cross-linking of SF, and the melt flow of the sample was inhibited. The thermal decomposition stability of fibroin molecules was greatly enhanced by iodine treatment.

Occurrence of a cathepsin B-like acid cysteine proteinase in the eggs of silkworm moth, Antheraea pernyi

In the crude extract of mature eggs of the pernyi silkworm moth, Antheraea pernyi, proteolytic activity was detected in the acidic pH region, with maximal activity at pH 3.5., however, no appreciable activity was observed in the neutral to alkaline pH region. The enzyme activity was strongly inhibited by cysteine proteinase inhibitors, such as iodoacetic acid (IAA) and N-[N-(L-3-trans-carboxyoxirane-2-carbony)- l-leucyl] agmatine (E-64); whereas, pepstatin, diisopropyl fluorophosphate (iPr 2P-F), and EDTA were without effect, suggesting the enzyme to be a cysteine proteinase. From the substrate specificity, major proteinase in the eggs is likely to be a cathepsin B-like proteinase. When the crude extract of eggs was incubated at pH 4.0, enzymatic activity appeared after a few minutes lag period, indicating that the enzyme was stored in a latent form in the eggs and thus was activated during the incubation. Pepstatin completely inhibited the activation but had no effect on the activated enzyme, suggesting that aspartic protease (s) was involved in this activation. The proteinase was found in vitro to readily degrade yolk polypeptides, as well as hemoglobin and other substrates. In the pupal ovary, this activity gradually increased during embryogenesis, reaching a maximum at days 10 after pupal ecdysis. After reaching a maximum, the level of cysteine proteinase remained approximately constant and the high activity was maintained during embryogenesis. The results altogether suggest that an acid cysteine proteinase (ACP) is stored as a latent form in the eggs and is implicated in yolk protein degradation of Antheraea pernyi eggs during embryogenesis.

Structure and physical properties of epoxide‐treated tussah silk fibers

AbstractThe structural characteristics and physical properties of epoxide‐treated tussah silk fibers from Antheraea pernyi silkworm are discussed in relation to the increasing weight gain values. Ethyleneglycol diglycidylether (E) and glycerin diglycidylether (G) were used as modifying agents. The noticeably high weight gain values (about 140%) obtained were attributed to the catalytic effect of SCN− anion absorbed by the fibers during the pretreatment under reduced pressure conditions. The amino acid analysis showed that epoxide G exhibited a slightly higher reactivity toward tyrosine, while arginine preferably reacted with epoxide E. The peak of loss modulus (E″) determined by dynamic viscoelastic measurements became broader and its position linearly shifted to lower temperature when the weight gain increased, and a minor peak appeared in the low‐temperature region below 50°C. Differential scanning calorimetry (DSC) thermograms showed that the position of the decomposition peak of modified silk fibers shifted to lower temperature with increasing weight gain values. The minor and broad endothermic peaks, appearing in the reference sample at about 234 and 290°C, disappeared by epoxide treatment. X‐ray diffraction patterns of tussah silk fibers suggested that the epoxide treatment does not affect directly the crystalline regions but causes a decrease of molecular orientation in the amorphous regions. Both briefringence (Δn) and isotropic refractive index (niso) of tussah silk fibers decreased by the reaction with epoxides, although with different rate and extent, confirming the decrease of average molecular orientation. The extent of decrease of strength and elongation depends on the kind of epoxide and on the weight gain value. Epoxide‐treated tussah silk fibers did not show significant changes of surface characteristics as the weight gain values attained up to 60%.