Silk fibroin (SF) produced by the domesticated wild silkworm, Samia cynthia ricini (S. c. ricini) is attracting increasing interest owing to its unique mechanical properties, biocompatibility, and abundance in nature. However, its utilization is limited, largely due to lack of appropriate processing strategies. Various strategies have been assessed to regenerate cocoon SF, as well as the use of aqueous liquid fibroin (LF_aq) prepared by dissolution of silk dope obtained from the silk glands of mature silkworm larvae in water. However, films cast from these fibroin solutions in water or organic solvents are often water-soluble and require post-treatment to render them water-stable. Here, we present a strategy for fabrication of water-stable films from S. c. ricini silk gland fibroin (SGF) without post-treatment. Aqueous ethanol induced gelation of fibroin in the posterior silk glands (PSG), enabling its separation from the rest of the silk gland. When dissolved in 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP), the SGF-gel gave a solution from which a transparent, flexible, and water-insoluble film (SGF_HFIP) was cast. Detailed structural characterization of the SGF_HFIP as-cast film was carried out and compared to a conventional, water-soluble film cast from LF_aq. FTIR and ¹³C solid-state NMR analyses revealed both cast films to be α-helix-rich. However, gelation of SGF induced by the 40%-EtOH-treatment resulted in an imperfect β-sheet structure. As a result, the SGF-gel was soluble in HFIP, but some β-sheet structural memory remains, and the SGF_HFIP as-cast film obtained has some β-sheet content which renders it water-resistant. These results reveal a structure water-solubility relationship in S. c. ricini SF films that may offer useful insights towards tunable fabrication of novel biomaterials. A plausible model of the mechanism that leads to the difference in water resistance of the two kinds of α-helix-rich films is proposed.

Although, the impact of mulberry leaves on mortality of Bombyx mori [silkworm] and cocooning rates has been investigated, limited research has exploited the influence on the quality of B. mori pupae. This study evaluated the effects of four mulberry cultivars (Ichinose, Noi-EX-Thailand, Ex-Thika and Victory1) on the nutritional quality of B. mori pupae. The proximate composition, amino acids, vitamins, and minerals of different mulberry cultivars and silkworm pupae were analyzed using standard methods. Silkworms fed on Noi-EX-Thailand and Ex-Thika cultivars exhibited the highest crude protein (57.9%) and fat content (44.3%), respectively. Eight essential amino acids were identified in silkworm pupae, with histidine (6.02–7.12 μg/100mg) being the most abundant, followed by significant amounts of lysine (1.40–5.03 μg/100 mg), leucine (1.96–2.47μg/100 mg), and valine (0.89–2.03 μg/100 mg). Pupae raised on Noi-Ex-Thailand leaves had the highest total amino acid content (23.01 μg/100 mg). Potassium was the major mineral in pupae (505.62–665.30 mg/100 mg), with the highest levels observed in those fed on Noi-EX-Thailand. Vitamin C levels ranged from 28.84 to 61.88 mg/100 g, with pupae fed on Victory1 showing the highest concentration. There was a strong positive correlation between magnesium, crude protein and nitrogen-free extracts of mulberry leaves that mirrored the corresponding levels in silkworm pupae. These results underscore the importance of mulberry leaf composition in determining silkworm pupae quality, especially when considering their use as sustainable ingredient for the food, feed and pharmaceutical industry.

Silk fibroin (SF) has attracted attention as a base biomaterial that could be suitable in many applications because of its shape and structure. Highly functional SF has been developed to promote tissue regeneration with heparin conjugation. However, the hydrophobic three-dimensional structure of SF makes it difficult to bind to high-molecular-weight and hydrophilic compounds such as heparin. In this study, sufficient heparin modification was achieved using tyrosine residues as reaction points to improve cellular response. As it was considered that there was a trade-off between the improvement of water wettability and cell responsiveness induced by heparin modification, influences on the structure, and mechanical properties, the structure and physical properties of the SF conjugated with heparin were extensively evaluated. Results showed that increased amounts of heparin modification raised heparin content and water wettability on film surfaces even though SF formation was not inhibited. In addition, the proliferation of endothelial cells and fibroblasts were enhanced when a surface with sufficient heparin assumed its potential in assisting wound healing. This research emphasizes the importance of material design focusing on the crystal structure inherent in SF in the development of functionalized SF materials.

Formation of the α-helical conformation in the poly-l-alanine (PA) sequence regions, subsequent structural transition to β-sheet during natural spinning, and presence of residual α-helices in Samia cynthia ricini (S. c. ricini) native silk fiber have been experimentally proven. However, the aggregation state of the residual α-helices, and their influence on the mechanical deformation behavior in native fiber remain unclear. Here we show that the α-helices form an ordered aggregation state with a hexagonal packing in the aqueous solution, some of which remain during natural spinning. X-ray scattering and differential scanning calorimetry (DSC) analyses revealed occurrence of a structural transition of the residual α-helices to the β-sheet structure, accompanied by disappearance of the plateau region in the force-strain curve, due to heat-treatment at ~220 °C. On the basis of X-ray scattering before and after tensile stretching of S. c. ricini native silk, a direct connection between the plateau region and the α-helix to β-sheet structural transition was confirmed. Our findings demonstrate the importance of the PA sequence regions in fiber structure formation and their influence on the tensile deformation behavior of S. c. ricini silk, features believed to be essentially similar in other saturniid silks. We strongly believe the residual ordered α-helices to be strategically and systematically designed by S. c. ricini silkworms to impart flexibility in native silk fiber. We anticipate that these knowledge forms a basis for fruitful strategies in the design and development of amino acid sequences for artificial silks with desired mechanical properties.

Over 60% of tea in Kenya is produced by smallholder tea farmers distributed in the East and West of the Rift Valley. With all smallholder tea farmers using the same production technologies including planting materials, the quality of tea from different agro-ecological zones should be the same. However, black teas attract different prices. This study compared levels of leaf quality, catechins, and polyphenols in green leaf and quality parameters of black tea from twelve factories in three different agro-ecological zones in Kenya and assessed the relationships between the green leaf quality parameters and black tea quality parameters. The catechins contents, the black tea quality parameters, and sensory evaluations changed (p less than or equal to0.05) between the factories and with an agro-ecological zone of production. All black teas had high antioxidant activities, which differed (p less than or equal to0.05) across the factories and zones. Despite the use of same agronomic inputs in the smallholder tea growing zones in Kenya, the quality of resultant black teas was largely influenced by the environmental factors in the different regions. It is therefore not possible to produce black teas of the same quality across the country. Despite the variations, all regions exhibited high levels of black tea quality parameters and sensory evaluation, re-affirming the Kenyan smallholder black teas are of high quality. The high antioxidant activities suggest possible beneficial pharmacological activities from consumption the teas. The green leaf quality (leaf count) were significantly (p less than or equal to0.05) correlated with black tea brightness and all sensory evaluation parameters, demonstrating high benefits from good plucking standards. Green leaf gallic acid, epigallocatechin, epicatechin, epigallocatechin gallate, and total catechins levels were related (p less than or equal to0.05) to black tea total polyphenols and all sensory evaluation parameters. In addition, the green leaf caffeine, epicatechin, epigallocatechin gallate, and total catechins influenced (p less than or equal to0.05) theaflavins in black tea. These green leaf parameters are therefore key drivers of Kenyan plain black tea quality.

Despite the use of similar cultivars in the smallholder tea sector in Kenya, resultant black tea quality varies. These variations could in part be arising from the formation of varying quantities and ratios of the black tea quality precursor compounds with agro-ecological zones of production. This study evaluated the variations in the caffeine and flavon-3-ol profiles of three cultivars (SFS 150, TRFK 31/8 and TRFK 303/577) of tea from smallholder tea farms in three agro-zones in Kenya. Overall, there were significant variations (p less than or equal to0.05) in the levels of caffeine and flavan-3-ols due to cultivars and agro-ecological zones of production. In the same cultivar, the components varied (p less than or equal to0.05) with an ecological area of production. The patterns in the changes were not systematically leading significant ecological zone x cultivar interactions effects in gallic acid, catechin, and epicatechin gallate. This demonstrated that it may not be predictable how the quality of clonal tea may vary when produced in different agro-ecological zone. Consequently, a high-quality clone in one agro-ecological zone may not replicate the same characteristics in the different agro-ecological zone. It is, therefore, necessary to test new clones in new environments before they are extensively exploited in these environments. The current findings suggest that flavon-3-ols may not be potential factors in the discrimination of tea quality within the ecological zones of Kenya.