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Caspian Journal of Environmental Sciences
The silk proteins, sericin and fibroin in silkworm, Bombyx mori Linn., - a review
M. Mondal*, K. Trivedy and S. Nirmal Kumar
Silkworm Physiology Laboratory, Central Sericultural Research and Training Institute Mysore, 570008, Karnataka, India. * Corresponding author’s E-mail: firstname.lastname@example.org
The domesticated silkworm, Bombyx mori Linn., a lepidopteran molecular model and an important economic insect that are emerging as an ideal molecular genetic resource for solving a broad range of biological problems. The silkworm, B. mori produces massive amount of silk proteins during the final stage of larval development. These proteins are stored in the middle silk gland and they are discharged through the anterior duct and spinneret, at the end of the fifth instar. Two kinds of silk proteins have been distinguished as major components of silk cocoons, the first being fibroin, a fibrous protein composed of heavy (H) chain, Light (L) chain and glycoprotein linked by disulfide bonds and the second being sericin a natural macromolecular protein, serving as an adhesive to unite fibroin for making silk cocoons of silkworm, B. mori. Recently, silkworm is being used as biofactory for the production of useful protein using the silk gland, which has promoted the technological development in sericulture. With the above background silkworm can be classified as a value added biomaterial for medical application, application of silk protein fibroin and sericin as a biomaterial and other seri-byproducts. The present paper overviews some important studies carried out on sericin and fibroin of silkworm, Bombyx mori Linn.
Keywords: Silk protein, Sericin, Fibroin, SDS polyacrylamide gel electrophoresis, Bombyx mori.
Insects mainly belong to two families, viz., Saturnidae and Bombycidae, which spins silk fibre. Bombyx mori belongs to Bombycidae produces a delicate twin thread of silk fibroin, which is coated by a protective cover of sericin. Silk protein is a kind of protein like collagen, elastin, keratin, fibroin, sporgin etc., is an essential constituent of cocoon filament (Komatsu, 1975). The silk fiber protein is synthesized by silk gland cells and stored in the lumen of the silk glands. Subsequently, it is converted into silk fibres. When the silkworms secrete the liquid silk during the spinning, it passes through the anterior gland and expelled out through the spinneret opening (Shimizu, 2000). Quantity and nature of sericin are fundamental characteristics in conferring distinctive traits to the cocoon (Sadov et al., 1987).
Sericin is insoluble in cold water, however, it is easily hydrolyzed, where by the long protein molecules brakes down to smaller fractions, which are easily dispersed, or solubilised in hot water (Gulrajani, 1988). Sericin protein is useful because of its special properties viz., resists oxidation, antib acterial, UV resistant and absorbs and release moisture easily, inhibitory activity of tyrosine and kinase etc. (Fig 1). Sericin, a major component of silk fiber, has been selectively removed from fibroin during the silk manufacturing process to make silk lustrous and the removed sericin goes as waste material. Now a days Seri- waste products and Seri- byproducts are used as a value added products. After Degumming, the leftover is fibroin made up of two brins. Silk fibre can be used for many purposes including textile, medical and industrial applications. The silk fibre is
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Ramification considerably enlarges the nuclear surface and apparently facilitates the transfer of materials related to the silk synthesis between the nucleus and the cytoplasm. which rival the most advanced synthetic polymers. According to its morphology and function. Phillips et al. It is a paired organ consisting of modified labial/salivary glands located at the two lateral sides under the alimentary canal. The anterior part about 2 cm long is a thin duct composed of about 250 cells with no known secretory function. In the mutant. a disulfide linkage between the heavy (H) chain and light (L) chain is not formed because of partial deletion of the L-chain gene. Diagrammatic representation of Attributes of sericin. light and soft Thermotolerance Fig 2. It is reported that the introduction of ionic liquids to silk processing opens an exciting avenue for controlling the microstructure to tune the macroscopic properties. If this sericin protein is recovered and recycled. sericin and fibroin in silkworm 250 to 300 tons of sericin per year (Gulrajani. Inoue et al. Diagrammatic representation of Attributes of Fibroin. Silk gland of B. mori is a typical exocrine gland secreting large amount of silk proteins. 00. (2005) reported that the Bombyx mori silk gland secretes one fibroin and three layers of sericin from the each posterior and middle silk gland in a normal larva. It is well known for its water absorbency. that have 50. It is the raw material for producing precious fabrics.64 thin. insulation properties and luster (Fig 2). about 15 cm long and is composed of about 500 secretary cells. (2005) investigated the possibility of using the NdsD mutant for the efficient production of recombinants in the silkworm. The posterior part. thermo tolerances. which is essential for the intercellular transport and secretion of fibroin. The posterior Anti Oxidant UV Protection SERICIN Moisturizer Coagulant Chemo Protective Nutrient Fig 1. SILK GLAND The natural silk synthesized by the silkworm and spun in the form of a silk cocoon is originally synthesized in the silk gland.000 tons of recoverable sericin. Indian production of 1. the protein which cements the fibroin thread of the cocoon. The cells constituting the gland are huge polyploid cells each with extremely ramified nucleus containing numerous nucleoli.600 tons of silk can be source of about . tyre lining materials. is about 7 cm long and contains about 300 secretory cells producing silk sericin. Nuclear ramification develops gradually as the larva grows and reaches conspicuous size in the 4th and 5th instars.. Water absorbency Insulation properties FIBROIN Dyeing affinity Lusture Thin. long. it would be a significant economic and social benefit. which synthesize silk fibroin. artificial blood vessels and surgical sutures.000 tons of dry cocoon are generated. Each gland is basically a tube made of glandular epithelium with two rows of cells surrounding the lumen. It is estimated that out of about 1 million tons (fresh weight) of cocoons produced world wide approximately 4. To utilize the inactivity of the mutant L-chain. 2005). parachutes. Akai et al. Proteins.. the silk gland can be divided into three distinct regions (Fig 3). long. light and soft. The Nd-sD mutant is silk fibroin secretion deficient. The silk fibers have outstanding natural properties. (2005) reported that ionic liquid could hold the key to the production of designer silk fibers with enhanced mechanical and optical properties. yet unlike synthetic polymers the production of silk does not require harsh processing conditions. dyeing affinity.. The middle silk gland in the lumen of which silk proteins are stored until spinning.
15 19.92 78.. B. although the fibroin of Saturnidae species secreted as dimer of Hchain. and Glycoprotein P25 30 k Da (Chevillard et al. 1982. mori consists of three protein component: High (H)-chain 350 k Da (Shimura et al. and P 25 is 6:6:1 for the fibroin secreted into the posterior silk gland (PSG).4-0.39 Layer 1 2 3 4 5 Fig 3. Zhou et al..Mondal et al.86 17. In order to use as a foreign proteins. Feiying et al. 1998).36 0. mori at different developmental stages.. but there is no such disorder in the middle silk glands.77 1. 1988). 1986a. L-chain.8 1.07 1. Low (L) .. Table 2.09 Sericin (%) 32. L-chain and P 25) are common among different silk producing insects in Lepidoptera.. Besides sericin.23 1. 2005)..41 23. These three types of fibroin (H-chain. Component Fibroin Sericin Wax matter Carbohydrates Inorganic matter Pigment Total % 70-80 20-30 0.6 0.15 1. Several promoters were tested to synthesize foreign proteins like procollagen III or globular ones and successfully produced in the silk glands of Bombyx mori (Chavancy. Sericin is chemically a non-filamentous protein.. Change in cocoon filament of a particular silkworm having dissimilar layers (Rui.chain 26 k Da (Yamaguchi et al. Fibroin (%) 64.7 0. COMPOSITION OF THE COCOON FILAMENT Gulrajani (1988) reported that the silk fiber is almost a pure protein fiber composed of two types of proteins viz.32 1.84 0. piggyBac vectors were developed to express transgenes in the silk gland using silk gene promoters to drive the expression of the integrated foreign genes.79 17. (2005) studied the analysis of protein variety of middle silk gland cells of the fifth instar larvae of silkworm.94 74.69 79. 2000). where in fibroin content is less (Table 2). fat and waxes. 2000). (2002) have introduced a new silkworm race which produces only sericin in Japan in the name of “Sericin Hope” introducing Nd-sD mutant. sericin and fibroin. raw silk also contain other natural impurities namely. Table 1. SILK PROTEIN FROM THE SILK GLAND Silk fibroin secreted in the lumen of posterior silk gland (PSG) of B. 1989). Yamamoto et al. inorganic salts and colouring mater (Table 1). 65 of the silk fiber and revealed that the sericin content is more in outer layer.. Composition of silk in Bombyx mori (Gulrajani. Quantitative enzyme linked immuneosorbent assay (ELISA) with specific antibody for each protein component showed that the molar ratio of H-chain.2-1.78 Ether soaked substance (%) 1.. Fibrohexamer was proposed as a functional name of P25 The silk gland has the capacity to produce large amount of silk proteins. Schematic representation of silk gland of silkworm (Bombyx mori L).2 100 silk glands are not sufficiently developed and the liquid fibroin is scarcely secreted.75 Ash content (%) 1.34 79. The N-linked oligosaccharide chain of P25 has been suggested to be involved in the later interactions (Inoue et al. 1986b). Rui (1998) studied the outer layer .09 1.
since the orientation is unstable by hot water treatment and thus there is a reversible relationship between the orientation and non-orientation. These MFFs adhered closely together showing a higher concentration in the posterior part of the middle silk gland. In the posterior part of the posterior silk gland. By the observation of various portion of silk gland. The structure of a crystalline form of B.. But the H-L linkage is essential for the large-scale production of fibroin (Mori et al. The (phi. sericin and fibroin in silkworm fibroin consists of non. When this film is swollen in water. The crystalline structure and molecular conformation of silk sericin obtained from wild silks is not practically different from that of sericin obtained from B. silk I. 1985). The disulfide linkage between Cys-172 of the L-chain and Cys-c20 of the H-chain (Tanaka et al. respectively.. Crystalline structure of the peptide chains in silk fibroin. The part of the liquid sericin in the middle silk gland that is easily crystallized by drying is possibly made of amino acid residues with short side-chains and is folded into the globular matrix made of stretches with longer side-chains. is determined by X. crystallizing less readily and on drying. . infrared spectrometry and differential scanning calorimetry of the wild silk (Antheraea pernyi. Antheraea yamamai. by means of X ray diffraction. (71°. According to Kenji et al. it changes to oriented fiber structure. become film of unoriented crystal structure... 2001). The crystalline region tends to be oriented along the fibre axis because the fibre is drawn as it is extruded from the spinnerets of the silkworm.66 and its central role is maintaining the structure of elementary unit.. 99°C) which are in the bridge and fourth quadrant regions of the Ramchandran map. it is concluded that the cocoon filament is composed of oriented elementary fibroin fibers and these fibers are derived from MFFs as they undergo structural change during the passage through the silk gland lumen. RHC C=O H-N CHR O=C N-H RHC C=O H-N C=O H-N O=C CHR H-N C=O RHC N-H O=C N-H RHC C=O H-N CHR CHR RHC C=O X . Tsukada (1983) studied the crystalline structure and molecular conformation of silk sericin. mori. 1999) is not responsible for the maintenance of the once formed elementary unit (Inoue et al. Proteins. Komatsu (1975) postulated molecular aggregating structure of sericin and its changes.ray diffraction method.crystalline and crystalline regions. mori.like structure by combining β obtained from solid-state two dimensional spin-diffusion nuclear magnetic resonance and rotationalecho double resonance (Asakura et al.RAY DIFFRACTION ANALYSIS Konishi (2000) studied the X ray diffraction of fibroin and noticed 1 mm thick parallel silk fibroin fibres. in order to analyze the mechanism involved in the formation of a single cocoon filament. However. 1995).. Philosomia cynthia ricini) by boiling in water. the cell dimensions are essentially the same as those found in the synthetic model polypeptide (LAla-Gly). pI) values of L-Ala and Glee in the repeating unit (-112°. Only difference is that the wild silk sericin is relatively insoluble compared to B. and become homogeneous and compact in the anterior silk gland. (2001). mori silk fibroin commonly found before the spinning process (known as silk I) has been proposed as a repeated β-turn type II. It is observed that H-N Fig 4. the columnar fibroin located in the lumen consisted numerous spherical masses of fibroin fibers (MFFs). mainly due to chemical interaction between silk sericin and inorganic minor components or tannins contained originally in wild silk. 2000). (1987) observed the fine structural change of liquid columnar fibroin in the lumen of silk gland during the passage from the posterior to the anterior silk gland during the spinning stage.-6°C). stretched and dried. Akai et al. The molecular and crystal structure of the crystalline modifications of B. because fibroin is retained in endoplasmic reticulum in the absence of disulfide linkage between the H and L-chains (Gamo and Sato. mori.
e. 1982). Thus the sericin gets modified in the direction of difficult solubility due to repeated moisture absorption and loss. but lacks α-helix. The transition of sericin from its random coil to β. Heating during moisture absorption activates the thermal motions of segments and accelerates transformation. most experimental evidences indicate that sericin exists mainly in the random coil or β-structure.In a new cycle of water absorption. rather than having the main axis of crystallites oriented at right angle to the fiber axis. a further fraction of random coils state crystalizes into a β structure there by increasing the portion of β structure and promotes crystalization.. fraction A can be separated by fractional precipitation by adding 15g of ammonium sulphate per 100 ml of sericin solution. hygroscopic conditions (Komatsu..shaped zigzag arrangement. Komatsu (1975) also reported that sericin. leading to remarkable agreement of observed and calculated structure amplitudes of both dipeptide and hexapeptide sequences. Sericin in aqueous solutions obtained on degumming silk is not an individual chemical substance but is a mixture of at least two substance. III to cover the fibroin. whereas . β-structure sericin is more insoluble than random coil sericin. 67 filament has molecular chains also arranged in cross β structure.. The intra. displays both random coil and βstructure. these reports are not in conformity with Tsukada (1983) based on analysis of circular dichroism spectrum.. MOLECULAR CONFORMATION OF SERICIN AND ITS TRANSITION Iizuka (1969) reported that sericin extracted from the liquid silk and fresh cocoon shells of a silkworm mutant. Ishikawa and Hirabayashi (1968) reported that sericins have a cross β structure when a water solution of sericin containing 50% dioxane (Iizuka. the crystalline structure already formed. Sadov et al. part of the sericin IV become a white suspension due to the coexisting cocoon yarn wax but does not coagulate and the β structure is originally present in the liquid silk. 1987). X ray structure analyses of the crystalline domains of fibroin show that the peptide chains pack in fully extended forms (Fig 4).molecular bonds of sericin having random coils are broken by the absorption of water molecules and the folded structure becomes unfolded into an extended structure and is transformed into a β structure.Mondal et al. the sericin layers are formed from the outside to inside in the order of I. II. Part of the sericin thus transformed into β structure is fixed by its new intermolecular hydrogen bonds and remains crystallized even when the water molecules are removed by drying. indicated that sericin extracted from liquid silk for 45 minutes with water contains a small fraction (10%) of α-helix. remains unaffected. The α-sericin contains lesser C and H and somewhat more N and O than the β-sericin (Bose et al.Sericin is present in the outer layer of cocoon shell and β-sericin in the inner layer. which. Some scientists described that on the cocoon thread. sericin A and Sericin B. Komatsu (1980) argued that during the dissolution of the liquid silk in water. sericin B will be precipitated. Three fractions of sericin which were different in solubility and called them sericin fractions I. .. (1987) have come to the conclusion that native sericin is mixture of two substances. β Structure is more stable regarding energy. 1969) or methanol (Ishikawa et al. Komatsu (1975) believed that the sericin fraction closest to fibroin in the cocoon SERICIN LAYER STRUCTURE OR SERICIN FRACTION Scientists have separated sericin of cocoon shell into two proportions: (1) α-sericin and (2) β -sericin. To date. which secretes only sericin. It is believed that β structure is intrinsic to liquid silk. α. is a random coil with 510 % β-structure and no α-helix. The solubility of α-sericin in the boiling water is more than β sericin. He studied the sericins from both the liquid silk in the silk gland and the cocoon filament and confirmed the relationship between solubility in hot water and molecular conformation (Komatsu .structure takes place by repeated absorption and de-absorption of moisture and by heating during the absorbed state i. 1989). and has a reasonable hydrogen bond networks. The observed molecular conformation has a “crank-shaft” or a S. in aqueous solution. When ammonium sulphate is added to the filtrate. Though. 1980).
and III. 1978). the sericin extracted from the floss showed high contents of serine.15 0.28 0. The greatest sericin content is present in the outer layer of cocoon whereas the least sericin proportion is present in the innermost layer of a cocoon. However.e.12 4.20 6.67 0. 78% of the weight of raw silk).70 5.64 2.60 0.09 0. galactosamine. III and IV by their different solubilities in hot water and assessing the degree of solubility by UV absorption. proline. cystine and phenyl amine in higher amount. glucosamine.03 Cocoon layer Outer layer 18.92 28.63 1.39 28. I. glycine. Fourth fraction of sericin was reported to have higher specific gravity and crystallinity than moisture content (Table 3).84 0.407 15.12 0.412 37.39 0.68 3.50 IV 3.60 Total sericin 100..90 0.30 8. cystine.26 0. galactosamine and histidine in lower amount and throeonine.58 0. 1975).99 3.10 0.29 40.76 16.20 1. galactosamine and glucosamine were significantly higher in the inner layer of cocoons than in the outer layer. aspartic acid.58 4. Sericin I Content (%) Co-efficient of dissolution velocity Moisture regain (%) Specific gravity Crystallinity (%) 41.53 16. Fibroin is the Proteins.70 15. It is found that of the sericin I. Further more.90 4.66 18.00 16.17 3.85 4. Properties of sericin fractionated with hot water (Komatsu. on the cocoon filament is amorphous where as II and III are crystalline.33 16. sericin layers can be called as Sericin A and Sericin B or Sericin and or Sericin I. mori. The sericin A is more soluble than the others.78 0. Amino acid Aspertic acid Threonine Serine Glutamic acid Proline Glycine Alanine Cystine Valine Methionine Isoleucine Leucine Tryrosine Phenylamine Lysine Histidine Arginine Inner 18.89 0.06 Table 4.79 0. AMINO ACID COMPOSITION IN SERICIN AND FIBROIN Yamada (1978) indicated that the sericin of mulberry wild silkworm.43 trace 3.89 0.43 1.50 1. the wild silkworm sericin contained serine. Bombyx mori mandarina. Table 3.94 3.67 0.46 9.47 2.56 16. methoinine.30 .91 0.80 5.51 16. valine and tyrosine but low contents in threonine.33 0.20 II 17.10 0. III and IV are having different solubilities harder to dissolve being sericin IV.11 0.28 4.22 14.17 4. B. sericin and fibroin in silkworm principal water insoluble protein (i. II.00 0. Based on the different degree of solubility.44 2. lysine and histidine as compared with the sericin of cocoon layer (Table 4).70 1. Amino acid composition of sericin obtained from cocoon layer and floss of the mulberry wild silkworm (Yamada.00 II 38.61 11.30 1. alanine. Komatsu (1975) observed that four fractions of sericin viz.68 II.46 0.400 3.42 2.403 18.03 3. leucine.22 10. and III based on the order of ease of dissolution in hot water and that their ratios were approximately 40:40:20.84 0.89 3.90 3..31 0. II.42 2. seem to contain the same kind of amino acids as the domesticated silkworm.05 4. Robson (1985) reported that sericin may be separated into sericin I. II. glutamic acid.44 29.41 Floss Whole 18.60 1.408 32.00 5.70 1.60 0.53 2.67 1. glucosamine.10 0. The content of threonine.
80 23. A small amount of cystine residues give a very small amount of sulphar in the fibre.45 7. implying that the transcription.87 28. Fibroin contains only a small amount of amino acids.69 25. which have acid side chains (Table 7).PAGE) OF SERICIN AND FIBROIN PROTEIN Kurioka and Yamazaki. The qualitative and quantitative differences in proteins expressed in the middle silk glands of male and female silkworm larvae that differential silk colour are investigated by high resolution twodimensional polyacrylamide gel electrop horesis (2-D PAGE). SDS POLYACRYLAMIDE GEL ELECTROPHORESIS (SDS . 69 significant difference in the protein composition of the same part of middle silk gland (MSG) cells among the second day. The wild silkworm sericin contained more amino acids with nonpolar side chain and less amino acid with polar side chain than the domesticated silkworm sericin (Table 5).31 43.3 and Mr 6000. Bombyx mori. Contents are express as mol %. Two dimensional polyacrylamide gel electrophoresis (2D-PAGE) resolved this protein into a single spot with pI 4. sixteen in the fourth day of fifth instar and twenty-four in the mature silkworm.49 42.92 41. Mulberry wild silkworm Groupof amino acid Inner cocoon layer 72.51 Floss 70.08 7.16 8.36 28. There is low proportion of amino acid residues with large chains in silk. thirty-three in the fourth day of fifth instar and thirty-four in the mature silkworm.43 Domestic silkworm Inner cocoon layer 76.25 40. Amino acid sequence analysis revealed that this protein consists of 55 amino acids.18 27. mori..32 2.65 Outer cocoon layer 71.26 28.35 Polar Hydroxyl Acidic Basic Nonpolar Ratio (Polar / Nonpolar) Values of the domestic silkworms are given as mean of 8 varieties. glycine and serine.. Feiying et al. 1980).66 14. seven in the fourth day of fifth instar and twenty-five in the mature silkworm. the fourth day and the mature silkworm of the fifth instar larvae of silkworm.38 Outer cocoon layer 77.36 2.88 23. Jain et al. Ten special protein spots are found in posterior part of MSG cell in the second day. (2002) purified protein with a low molecular mass of 6027 from the cocoon shell of silkworm. translation of fibroin gene may involve a complex regulation system with great deal of regulation points. Fibroin has high proportions of alanine. (2005) analyzed 2DPAGE patterns of protein from posterior silk gland of 4th day silkworm in 5th instar of different breeds of B. Content of amino acids with polar and nonpolar side chains constituting in the cocoon sericins from wild and the domestic silkworm (Yamada.51 6.75 3.Mondal et al. The result indicated a Table 5.46 21. The co-efficient of pattern-similarity in amino acid composition was high between the sericin of wild silkworm and domesticated silkworms. The isoelectric point of silk fibre is around pH 5. The results show that the amino acid composition of sericin extracted from the cocoon is species specific. Nine special protein spots are detected in the middle part of MSG cells in the second day. Eight special protein spots are expressed in anterior part of MSG cells in the second day. Protein forms of posterior silk gland are apparently different for different breeds and different protein spots are found when they compared to middle silk gland of different breed. six of these being cystine residues and is highly homologous to bovine pancreatic trypsin inhibitor type.78 3.79 2.04 50.51 5. while sericin of the wild silkworm such as Antheraea or Phylosamia showed significantly low similarity (Table 6). (2005) uses the two dimensional Polyacrylamide gel electrophoresis (2D-PAGE) and the image analyses technology.limited character. . They reported that there are no distinct differential proteins between the silk gland tissue of female and male larvae within the same variety of sex.11 22.
3 3.1 1.6 121. Pattern. Like other common biomedical textiles such as polyester. APPLICATION OF FIBROIN PROTEIN SERICIN AND Isolation of the smallest component of silk Tokutake (1980) reported a series of polymers of the smallest component. The smallest component in the sericin fraction is purified by re-chromatography and showed a single band on gel electrophoresis. . serine.friendly biodegradable polymers can be produced by blending sericin with other resins (Annamaria et al. Thus fibroin and sericin can be separated by precipitation then both the fraction of silk can be examined by polyacrylamide gel electrophoresis. 1978).06 M-cupric hydroxide) solution showed distinct bands on polyacrylamide gel electrophoresis in the presence and absence of SDS. which was rich in glycine. Amino acid composition of silk fibroins (residues/ 1000 residues) (Robson 1985).4 51. On the gel electrophoresis.B i i cos θ = i =1 n n ∑ Ai 2 i =1 ∑B i =1 i2 Table 7.0 5. and that there exists a series of polymers.946 0. Fibroin and sericin fraction were Silkworm is being used as biofactory for the production of useful protein.977 0. assamensis Philosomia cynthia ricini Pattern-similarity coefficient=S (A.0 10. On the other hand.7 1.990 0. sericin showed distinct band but fibroin did not.974 0.13 M Ethylenediamine/ 0. B ) = n Proteins. mori fibre 446. detected by polyacrylamide gel electrophoresis.976 ∑ A . Silkworm Bombyx mori mandarina Bombyx mori Antherea yamamai Antheraea pernyi A.000 0.0 9. 1998). the combined contents of which amounted to 72mol%. Biodegradable materials Environment . Silk proteins are natural polymers and are biodegradable with reactive functional groups that open up possibility to be crosslinked with other polymers to be used in controlled delivery. alanine and aspertic acid. sericin and fibroin in silkworm Pattern-similarity coefficient 1. silk contains various polar functional groups that might enhance antibiotic absorption.0 2. paphia mylitta A.70 Table 6. These polymers are reduced by 2mercaptoethanol to the smallest component of silk protein. 1980).2 3.. could be converted into the smallest component by reduction and aminoethylation.1 13. the precipitate fraction is known as sericin.970 0. The component of the fibroin and sericin are fractionated by gel filtration on sepharose 6B. The supernatant fraction known as fibroin is particularly rich in glycine and alanine. Amino acids Glycine Alanine Valine Leucine Isoleucine Serine Threonine Aspartic acid Glutamic acid Lysine Arginine Histidine Tyrosine Phenylalanine Proline Tryptophan Methionine (Cysteine)2 B.7 6. The results of electrophoresis suggest that components of sericin have define molecular weights but those of fibroin do not.6 1.5.2 4. which together accounted for 78-mol%. which had an apparent mol wt of 24000 (Tokutake.0 294.3 6. The amino acid compositions of two fractions are shown in Table 8.similarity coefficient of amino acid composition of the sericin from mulberry wild silkworm (A) and other silkworms (B) (Yamada. Silk protein solution prepared by solubilization of a whole cocoon with ED/Cu (0.0 22.0 Gly>Ala separated by precipitation of sericin at pH 5.
49 14. the bioprocessing industry and some chemical processes.22 4. dialysis. and fibers containing sericin (0. foams.50 5.99 4.21 18. Sericin.04 0.03 1.00 Precipitate Fraction Sericin 12. 71 Table 8. Sericin and fibroin can be used to make membranes for use in separation processes.01-50% w/w) can be produce by reacting a composition comprising a polyol.69 24.55 Contents are express as mol %. Use of the coated sericin film is an effective antifrosting method that can be .28 0. Other procedures have also been reported for producing sericin-containing polyurethane with excellent mechanical and thermal properties (Hatakeyama.6 2.10 0. or copolymerized with other substance are Functional biomaterials Nakajima (1994) has found that sericin film located on lay of liquid crystal can uniformly orient the liquid crystal molecules to provide distortion-free high. Yamada et al. reverse osmosis.isocyanet. di. Membrane materials Membrane based separations (e. 1996).50 2.54 0. The insolubilized silk fibroin membrane could be used to separate the mixture of water and alcohol (Chisti. blended.9 5.69 2. (1991) describe a cross.70 2... di-butyltin di-laurate (catalyst) and trichloromonofluoromethane (a blowing agent) in the presence of sericin.08 0.2 15..58 0.25 0. tolylene.07 0.96 0.82 1. Supernatant Fraction Fibroin 1. The Polyurethane foams incorporating sericin are said to have excellent moistureabsorbing and desorbing properties (Minoura et al. 1980).7 13.. production of extremely pure water.66 12. Acrylonitrile used in making certain synthetic polymers can be copolymerized with sericin to prepare a protein containing synthetic polymer film for separating water from organics (Yamada and Fuwa 1994.45 3. but membranes of sericin cross-linked.2 3..00 0.17 9.9 4. Amino acid composition of supernatant fraction. Sericin containing membranes are quite hydrophilic. made readily.11 1.76 0.46 2.00 5.g.29 0.4 31.linked thin film made of sericin for use as a separating membrane for water and ethanol.coated film is used on the surface of refrigeration equipment because of its antifrosting action (Tanaka and Mizuno.98 3. Pure sericin not easily made into membranes. the precipitate fraction and the smallest component of silk protein (Tokutake. because sericin contains large amount of amino acid with neutral polar functional groups. 1995).00 46.quality crystal displays. ultra filtration and micro filtration) are used in process such as desalination of water. 1993).0 7.34 0.02 Trace Proteins Aspartic acid Threonine Serine Glutamic acid Proline Glycine Alanine Valine Methionine Isoleucine Leucine Tryrosine Phenylamine Lysine Histidine Arginine AminoEthylcysteine Smallest component 15. molding resins.68 1.98 0.2 3.45 3.00 0.2 5.Mondal et al. Mizoguchi et al. 2001). 1998).48 0. The moister absorption/desorption rate of the sericin containing polyurethane form is two-to five fold grater than that of control. Polymer films.
smooth. foam-forming aerosol shaving gels. The silk sericin has resemblance with the natural moisturizing factor (NMR). 2001). (1999) investigated the attachment and growth of animal cells on films made of sericin and fibroin. sericin-coated powders for cosmetics. Kato et al. Subsequently. additives to rice cooking. silk fibres have proven to be effective in many clinical applications. (1998) provided the first evidence of antioxidant action of the silk protein by showing that sericin suppressed in vitro lipid peroxidation. etc. as additives for health foods to prevent colon cancers.72 widely applied to refrigerators. some biological responses to the protein have raised questions about biocompatibility. hygroscopic moisture-releasing polyurethane foams and their manufacture for furniture and interior materials.linking agent. sericin and fibroin in silkworm Medical biomaterials Silkworm silk fibers have been the primary silk.fibroin blended film could be used to form article corneas (Murase. deep freezers and refrigerated trucks and ships. Sericin gel is prepared by using sericin solution with pluronic and carbopol as a stabilizer to prevent water loss from the upper layer of the skin.. The material coated the sericin have excellent weather ability. Silk protein can be made into a biomaterial with anticoagulant properties. Miyairi and Sugiura (1978) reported a cross-linked sericin film for enzyme immobilization with glutaraldehyde as the cross. coagulant for purification of waste waters. which brings wrinkles and dark spots. the electro-osmosis resistance and the stability of the immobilized β-glucosidase on the cross-linked sericin film are higher than the free enzyme. Sericin protein can be coated on surfaces of various durable materials to enhance functionality (Li.100 µm. The heat stability. especially with polyvinyl alcohol (PVA). (1995) and Tsukada et al. Moreover use of the coated film on roads and roof can prevent frost damage. These results suggested that sericin is the valuable natural ingredient for food and cosmetics. A membrane composed of sericin and fibroin is an effective substrate for the proliferation of adherent animal cells and can be used as a substitute for collagen. Furthermore. 1997). long lasting. Tasubouchi (1999a) developed a silk fibroin. At the same time..based wound dressing that could accelerate healing and could be peeled off without damaging the newly formed skin. artificial skin. The other uses of sericin includes. The non-crystalline fibroin film of the wound dressing had a water content of 3-16% and a thickness of 10. Excessive transepidermal water loss (TEWL) is one of the causes of dry skin and skin moisturizers have been used to overcome it. good permeability and do not warp on drying.. 2000). The use of oxygen-permeable membranes from silk fibroin and silk sericin. Sericin blends with water. Proteins. fabric care compositions. It is discovered that sericin can restrain the functions of active-oxygen (major factor of aging). A blended hydrogel made of sericin and fibroin and PVA is said to have excellent moisture absorbing and desorbing properties and elasticity (Yoshii et al. Film made of sericin and fibroin has excellent oxygen permeability and is similar to human cornea in its functional properties. by a sulfonation treatment of sericin and fibroin (Tamada.. 1994). semi-occlusive. Cell attachment and growth were dependent on maintaining a minimum of around 90% sericin in the composite membrane. A novel mucoadhesive polymer has been prepared by template polymerization of acrylic acid in the presence of silk sericin (Ahn et al. During decades of use. Minoura et al. 1999b). 1996). 2005).. It forms a moisturizing. medical composites of sericin. as dermatitis inhibitor.soluble polymers. the wound dressing was made with a mixture of both fibroin and sericin (Tsubouchi. antiwrinkle film on the skin surface imparting an immediate. sericin also found to inhibit tyrosinase activity. light and sunscreen comp ositions. containing about 60% water for contact lens. as wound protection . Sericin can be used in preparation of art pigments and for surface protection of articles. The biopolymer sericin has a strong affinity to keratin. protective.like material used in biomedical applications particularly as sutures. as a soil conditioner. That is why sericin can naturally saturate into skin and revitalize cells. silky feeling (Padamwar et al. It hoped that the sericin. The configuration of sericin is very close to the one of human beings. The hydrogel can be used as a soil conditioner and in medical materials and wound dressing.
274–280.Mondal et al. Asakura. J. (1996) studied the properties and application of wound protective membrane made by silk fibroin..B. S. K. seric. T. I. Y. and Orio... to controlled release type dosage tablets is investigated in vitro and in vivo. 2005). Nagashima. apparently due to interference with the adsorption of virus particles to CD4+ cells.. 119. Using fibroin controlled release tablets. 2004). Dandin. Biopolymers. 1989). Int. a major silk protein. and Ando.. optical beauty. The aqueous solution of fibroin is used to prepare a membrane for immobilization of Aspergillus niger glucose-oxidase and Pseudomonas fluorescens lyophilized cells (Demura et al.. Majumdar. Silk protein sericin. (1994) Investigation of uranium recovery from dilute aqueous solutions using silk fibroin. Inoue. Nakazawa..Fibroin has been explored as a biomedicine for various applications. (1994) investigated the uranium recovery from dilute aqueous solutions using silk fibroin. Akai. K.. 58. The applicability of fibroin. Hu.K. 43. Mysore. nail cosmetics. Jpn. It is concluded that the fibroin membrane has good wound healing properties. 131-137. Biol. Res.. (1987) Fine structural changes of liquid silk in the silkgland during the spinning stage of Bombyx mori larvae. Sericin and fibroin have been recently explored in the field of drug delivery systems. Solution blending is used to incorporate RHLC with silk fibroin to enhance the blend films biocompatibility ACKNOWLEDGEMENT The authors are thankful to Dr. C. Bangalore. M. M. S.. Kobayashi. lipstick and nail enamel. Silvio.H. The sulfated silk fibroins have anti-HIV-1 activity in vitro. (2001) Novel mucoadhesive polymer prepared by template polymerrization of acrylic acid in the presence of silk sericin. Choi. 80. 737-743. Biodeterior. J. Akai.H. Lee. H. film. (2001) Structure of Bombyx mori silk fibroin before spinning in solid state studied with wide angle Xray scattering and 13 cross polarization/ magic angle spinning NMR. and chewing gums (Gulrajani. India. Trace Elem. 20th congress of the international sericultural commission. The extremely fine powder (11. H. and Sengupta.C. Maria. Wu et al. 73 and hydrophilicity while maintaining elasticity. J.. and completely blocked virus binding to the cells at a concentration of 100 micro gm/ml (Gotoh et al.S.A. gels and mesosphere have been prepared. and Tsubouchi. REFERENCES Ahn. Yamare. 42.. Biodegrad. Fibroin powder was processed in such a way to retain its natural. P. (1989) Role of the amino acids in . The fibroin hydrogels prepared either by treating a 2% (w/v) silk fibroin aqueous solution at 4 °C temperature or by adding 30% glycerol could be used as scaffolds able to promote in situ bone regeneration (Matta et al.. The fibroin protein is one kind of biological materials used for artificial skin and others medical application.. and Cho.T. Sci. R. pp. 56.. Soluble fibroin enhances Insulin sensitivity and glucose metabolism in 3T3-L1 Adiposities. S. Imai. M. 2003). Bose. 2000).. India for encouragement. 1998)... Aslani and Eral. Tullia. Appl.. Aslani. Kamada..K. eye make up. S.3 µ size) is particularly ideal for applications in pressed powders. J. Sci. T. and Eral. K. (2005) Functional recovery of transgenic silk gland.. Central Sericultural Research and Training Institute. H. Yoshimura et al. K. S. blusher. T. T. Annamaria. Nahm. The silk fibroin can be used as the substratum for the culture of animal cells in place of collagen (Inouye et al. T. Polym. T. 521-525. (2006) reported that the Recombinant human-like collagen (RHLC) is blended with fibroin to prepare a novel biocompatible film as a scaffold material for hepatic tissue engineering applications. (1998) The microbial degradation of silk: a laboratory investigation. 1518th December. and Tarmura.. 203– 211. A unique property of this silk powder is its ability to hold and release moisture depending on the temperature and humidity of the surroundings. (1989) reported that the fibroin membrane is used to immobilizing coenzed insect cell culture as a vaccine. suppress DMBA-TPA induced mouse skin tumor genesis by reducing oxidative stress. Silk fibroin membrane supports the application as photo sensor for hydrogen peroxide analysis. Director. inflammatory responses and endogenous tumor promoter TNF-alpha (Zhaorigetu et al. S.
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