Simultaneous Extraction of Oil and Protein from Silkworm (Bombyx mori L.) Pupae (Lueng Parroj var.) and Their In Vitro Skin Moisturization.

Oil and protein from silkworm (Bombyx mori var. Leung Pairoj) pupae, by-product from sericulture, were extracted and evaluated for their potential uses as skin biomoisturizer. The silkworm pupae (SWP) oil and protein were simultaneously extracted by using three-phase partitioning (TPP) method and determined for their physicochemical properties including fatty acid and amino acid content, respectively. The highest yields of oil and protein at 8.24 ± 0.21% and 8.41 ± 0.26% w/w, respectively were obtained from 18 h extraction. Fatty acid analysis of SWP oil was rich in linolenic acid (37.81 ± 0.34%), oleic acid (28.97 ± 0.13%), palmitic acid (21.27 ± 0.05%), stearic acid (6.60 ± 0.09%) and linoleic acid (4.73 ± 0.21%). The clear yellow SWP oil possessed saponification value of 191.51 mg/g, iodine value of 119.37 g I2/g and peroxide value of 2.00 mg equivalent O2/kg. The SWP protein composed of 17 amino acids which aspartic acid, glutamic acid, glycine and serine were the major residues. SDS-PAGE analysis revealed that the SWP protein consisted of distinct protein at around 51, 70, 175 and over 175 kDa. Cytotoxicity of the SWP oil and protein was evaluated by using MTT assay and they showed low cytotoxicity toward keratinocyte cell (HaCat cell line). The SWP oil provided moisturizing effect on pig skin comparable to olive oil, while 1% and 2% of SWP protein showed higher moisturizing efficacy than 3% hydrolyzed collagen. The study indicated that the SWP oil and protein could be potential biomoisturizers for cosmetic products.

Amino acids imprinted ZIF-8s for the highly efficient and selective adsorption of antioxidant peptides from silkworm pupa protein.

Antioxidant peptides from edible protein sources have attracted great attention due to their high activity, easy adsorption and low toxicity. The purpose of this study was to develop a new MOF material with specific recognition and capture ability to achieve the aim of highly efficient and selective antioxidant peptides from silkworm pupa protein hydrolysate-N (SPPH-N). Firstly, Histidine (His) widely presented in antioxidant peptides was used to in-suit construct the modified ZIF-8 material (ZIF-His) with imprinted defective mesopores, based on "molecular imprinting strategy". Then, it was applied to extract antioxidant peptides from SPPH-N. ZIF-His exhibited high adsorption uptake for antioxidant peptides and especially presented specific adsorption property for peptides contained His, tryptophan, phenylalanine and tyrosine. Three peptides with the highest antioxidant activity (FKVPNMY, AVNMVPFPR and VNMVPFPR) from ZIF-His eluent were identified by Nano-LC-MS/MS. These peptides were synthesized and presented the excellent free radicals scavenging activity and the Ferric reducing power.

Effect of Drosophila suzukii on Blueberry VOCs: Chemical Cues for a Pupal Parasitoid, Trichopria anastrephae.

Biocontrol agents such as parasitic wasps use long-range volatiles and host-associated cues from lower trophic levels to find their hosts. However, this chemical landscape may be altered by the invasion of exotic insect species. The spotted-wing drosophila (SWD), Drosophila suzukii (Diptera: Drosophilidae), is a highly polyphagous fruit pest native to eastern Asia and recently arrived in South America. Our study aimed to characterize the effect of SWD attack on the volatile organic compounds (VOCs) of blueberries, a common host fruit, and to correlate these odor changes with the olfactory-mediated behavioral response of resident populations of Trichopria anastrephae parasitoids, here reported for the first time in Uruguay. Using fruit VOC chemical characterization followed by multivariate analyses of the odor blends of blueberries attacked by SWD, we showed that the development of SWD immature stages inside the fruit generates a different odor profile to that from control fruits (physically damaged and free of damage). These differences can be explained by the diversity, frequency, and amounts of fruit VOCs. The behavioral response of T. anastrephae in Y-tube bioassays showed that female wasps were significantly attracted to volatiles from SWD-attacked blueberries when tested against both clean air and undamaged blueberries. Therefore, T. anastrephae females can use chemical cues from SWD-infested fruits, which may lead to a successful location of their insect host. Since resident parasitoids are able to locate this novel potential host, biological control programs using local populations may be plausible as a strategy for control of SWD.

Nutritional targeting modification of silkworm pupae oil catalyzed by a smart hydrogel immobilized lipase.

To prepare a nutritional supplement using silkworm pupae oil (SPO) as a feedstock, a microfluidic reactor with a smart hydrogel immobilized lipase was first constructed to reduce the relative content of palmitic acid at sn-1,3 and improve the nutritional function. The effects of flow rate, reaction temperature, and substrate molar ratio were investigated. In vitro digestion and pH-stat models were employed to analyze the digestion feature after the modification of SPO, while HPLC-ELSD, zeta potential, DSC, and TGA were used to evaluate the nutritional function. The relative content of "OOO" and "OPO" type triglycerides was increased by 49.48% and 107.67%, and that of palmitic acid at sn-1,3 was decreased by 49.61% in 10 s. After the verification of the in vitro digestion model, the fatty acid release rate of the modified SPO was significantly improved by 22.07%, indicating the nutritional function improvement of SPO. Therefore, the nutritional function of SPO has been improved successfully by the application of a microchannel reactor with photo-immobilized lipase, which could set a reference for the utilization of insect oil resources.

Effect of the silkworm pupa protein-glucose conjugate on the thermal stability and antioxidant activity of anthocyanins.

Anthocyanin (cyanidin-3-O-glucose) is a natural water-soluble pigment with a robust antioxidant capacity. However, its poor stability and bioavailability limits its application as a functional food ingredient. This study explored the ability of the silkworm pupa protein-glucose (Spp-Glu) conjugate, developed under wet-heating conditions, to improve the thermal stability and antioxidant activity of cyanidin-3-O-glucose (C3G) at pH 3.0 and 6.8. The characterization experiments suggested that C3G complexed with the Spp-Glu conjugate could modify the protein's microenvironment and cause unfolding of the protein's secondary structures under varied pH conditions. Spectroscopic techniques further revealed the formation of complexes via hydrophobic interactions and static quenching processes when C3G was bound to Spp or Spp-Glu. The formation of these complexes effectively attenuated C3G degradation, thereby enhancing its stability under heat treatment over a range of pH values, and the experiments measuring antioxidant activity suggested that the Spp-Glu conjugate formed does not affect the efficacy of C3G after complexation. Therefore, our study suggests that Spp-Glu has the potential to effectively protect and deliver anthocyanins during industrial application for functional food formulation.

Purification and Identification of Novel Antioxidant Peptides from Enzymatically Hydrolysed Samia ricini Pupae.

The emergence of excessive free radicals leads to the destruction of various systems within the body. These free radicals also affect nutritional values, color, taste, and emit an odor akin to rancid food. Most food industries use synthetic antioxidants, such as BHT (butylated hydroxytoluene) or BHA (butylated hydroxy anisole). However, high doses of these can be harmful to our health. Therefore, an antioxidant compounds, such as bioactive peptides from edible animals or plants, have emerged to be a very promising alternative as they reduce potential side effects. This study focused on the purification and identification of antioxidant peptides from protein hydrolysates of wild silkworm pupae (Samia ricini). Antioxidant peptides were purified from the hydrolysate by ultrafiltration and RP-HPLC. The results showed that protein hydrolysate from S. ricini pupae by trypsin with a molecular weight lower than 3 kDa and highly hydrophobic property, exhibited strong DPPH radical scavenging activity and chelating activity. Further identification of peptides from the fraction with the highest antioxidant activity was carried out using LC-MS/MS. Three novel peptides, i.e., Met-Ley-Ile-Ile-Ile-Met-Arg, Leu-Asn-Lys-Asp-Leu-Met-Arg, and Glu-Asn-Ile-Ile-Leu-Phe-Arg, were identified. The results of this study indicated that the protein hydrolysate from S. ricini pupae possessed potent biological activity, and the novel antioxidant peptides could be utilized to develop health-related antioxidants in food industry.

An alternative solution for α-linolenic acid supplements: in vitro digestive properties of silkworm pupae oil in a pH-stat system.

α-Linolenic acid (ALA) is recognised to have a regulatory effect on cardiovascular diseases. Due to the low bioavailability of linseed oil (LINO), which is the most common ALA supplement, it is necessary to find a replacement for ALA supplements that is more easily accepted by the human body. The content of ALA in silkworm pupae oil (SPO) is 32.60 ± 0.67%, and SPO can be substituted as a dietary lipid to meet the demand of the human body. In the present study, a pH-stat system was used to investigate the release degree of free fatty acids (FFAs) from SPO and construct a first-order kinetic model. Digestion experiments in vitro with different lipids showed that the maximum release FFA levels were SPO > SO (soybean oil) > LO (lard oil) > MSO (mulberry seed oil) > LINO, and the first-order kinetic apparent rate constants were LINO > SPO > LO > SO > MSO. Triacylglycerol (TAG) and fatty acid composition are the decisive factors in determining the level of lipid digestion. Therefore, the maximum level of FFAs released from SPO (84.34 ± 1.37%) was much higher than that of LINO (49.78 ± 0.52%) when the hydrolysis rates were 0.2114 s-1 and 0.2249 s-1, respectively. In addition, the smaller emulsion droplet size (609.24 ± 43.46 nm) and weaker surface charge (-17.93 ± 0.42 mV) also resulted in higher levels of SPO under in vitro digestion conditions. Meanwhile, due to low melting and crystallisation temperature, SPO is quickly absorbed by the human body. Overall, SPO can be used as a new alternative for ALA supplements based on its superior digestive properties.

Two-step enzymatic synthesis of α-linolenic acid-enriched diacylglycerols with high purities from silkworm pupae oil.

In this study, α-linolenic acid-enriched diacylglycerols (ALA-DAGs) were prepared via a two-step enzymatic way by combi-lipase using silkworm pupae oils as substrates. Firstly, several factors including temperature, mass ratio of water to oil, pH and enzyme loading were optimized for the hydrolysis of silkworm pupae oil. The maximum fatty acid content (96.51%) was obtained under the conditions: temperature 40 °C, water/oil 3:2 (w/w), pH 7, lipase TL100L loading 400 U/g, lipase PCL loading 30 U/g. Then, ALA was enriched by urea inclusion, with an increased ALA content of 82.50% being obtained. Secondly, the ALA-enriched silkworm pupae DAG oil (SPDO) was prepared by lipase PCL-catalyzed esterification reaction. After molecular distillation, the final SPDO product contained contents of DAGs (97.01%) and ALA (82.50%). This two-step enzymatic way for production of ALA-DAGs was successfully applied in a 100-fold scale-up reaction. Overall, our study provides a promising way for the preparation of ALA-DAGs.

Changes in composition and levels of hemolymph proteins during metamorphosis of Manduca sexta.

The tobacco hornworm, Manduca sexta, is a lepidopteran model species widely used to study insect biochemical processes. Some of its larval hemolymph proteins are well studied, and a detailed proteomic analysis of larval plasma proteins became available in 2016, revealing features such as correlation with transcriptome data, formation of immune complexes, and constitution of an immune signaling system in hemolymph. It is unclear how the composition of these proteins may change in other developmental stages. In this paper, we report the proteomes of cell-free hemolymph from prepupae, pupae on day 4 and day 13, and young adults. Of the 1824 proteins identified, 907 have a signal peptide and 410 are related to immunity. Drastic changes in abundance of the storage proteins, lipophorins and vitellogenin, for instance, reflect physiological differences among prepupae, pupae, and adults. Considerably more proteins lacking signal peptide are present in the late pupae, suggesting that plasma contains relatively low concentrations of intracellular components released from remodeling tissues during metamorphosis. The defense proteins detected include 43 serine proteases and 11 serine protease homologs. Some of these proteins are members of the extracellular immune signaling network found in feeding larvae, and others may play additional roles and hence confer new features in the later life stages. In summary, the proteins and their levels revealed in this study, together with their transcriptome data, are expected to stimulate focused explorations of humoral immunity and other physiological systems in wandering larvae, pupae, and adults of M. sexta and shed light upon functional and comparative genomic research in other holometabolous insects.

Pupae protein extracts exert anticancer effects by downregulating the expression of IL-6, IL-1ß and TNF-α through biomolecular changes in human breast cancer cells.

The Pupae of Bombyx mori and Samia ricini are a source of high-quality proteins and essential nutrient elements for human. Recent studies revealed that protein extracted from pupae possessed therapeutic benefits for the treatment of many diseases. However, the anticancer activity of protein extracts from the pupae of B. mori and S. ricini has been rarely reported. Our objective was to study the effect of protein extracts from the pupae of B. mori and S. ricini on cytotoxicity and expression of pro-inflammatory cytokines; IL-6, IL-1ß and TNF-α, in breast cancer cells (MCF-7). Additionally, anticancer action of protein extracted from the pupae was further investigated through biomolecular changes in MCF-7 cells using Fourier transform infrared (FTIR) spectroscopy. Pupae protein extracts of B. mori exhibited cytotoxic effects with an IC50 value of 15.23 + 0.4 µg/mL with higher selectivity than doxorubicin on MCF-7 cells. Fourier transform infrared (FTIR) spectroscopy revealed that lipid contents in MCF-7 cells treated with pupae protein extracts of B. mori were higher than untreated cells. Treatment with protein extracts from pupae of B. mori or S. ricini caused significantly reduced protein and nucleic acid contents of MCF-7 cells. The expression of IL-6, IL-1ß and TNF-α in MCF-7 treated cells was investigated using RT-qPCR and ELISA. Our results revealed that protein extracts from the pupae of B. mori or S. ricini significantly decreased IL-6, IL-1ß and TNF-α in MCF-7 cells both at mRNA and protein levels. Expression of IL-6 and IL-1ß in MCF-7 treated cells, especially IL-6, was strongly reduced compared to untreated cells, while TNF-α expression was slightly decreased. These findings suggest that pupae protein extracted from B. mori or S. ricini may play a role in breast cancer through a down-regulatory action on the expression of IL-6, IL-1ß and TNF-α, and may also exert anticancer effects by causing biochemical changes of lipids, proteins and nucleic acids. These findings indicate that pupae protein extracted from B. mori or S. ricini may provide a potential novel therapeutic target for breast cancer.

Regulation mechanism of silkworm pupa oil PUFAs on cholesterol metabolism in hepatic cell L-02.

BACKGROUND: Silkworm pupa oil polyunsaturated fatty acid (SPO PUFA) has been confirmed to have a cholesterol-lowering function. METHODS AND RESULTS: The effect of SPO PUFA and its main component, α-linolenic acid (ALA), on the metabolism of cholesterol and its regulation was investigated. The model of lipid denatured cells were constructed to carry out lipid accumulation, cholesterol metabolism and transformation. Real-time PCR and western blots were also used to analyze the expression levels of related genes and proteins to investigate the cholesterol efflux regulation mechanism. The data indicated that SPO PUFA and ALA dose-dependently decreased intracellular total cholesterol (TC) and enhanced total bile acid (TBA). They could also promote cholesterol removal by enhancing bile acid secretion and by upregulating genes LXRα, PPARγ, ABCA1, ABCG1, and CYP7A1, which were regulated by LXRα/PPARγ-ABCA1/ABCG1-CYP7A1 nuclear receptor signal pathways. CONCLUSIONS: This study is of great significance in maintaining the balance of cholesterol and lipid metabolism, and in reducing the risk of steatohepatitis. © 2019 Society of Chemical Industry.

Promotion of Metabolite Synthesis in Isaria cicadae, a Dominant Species in the Cicada Flower Microbiota, by Cicada Pupae.

Cicada flowers, which are edible and medicinal mushrooms, are the fruiting bodies of Isaria cicadae, a fungus that is parasitic on the larvae of cicada pupae. We hypothesize that host factors might possess stimulatory activity on metabolite synthesis in Isaria cicadae. Here, we first compared the microbial community structures of different wild cicada flowers across geographical regions, compartments, and growth stages via high-throughput sequencing. Isaria cicadae TZC-3, an isolate of the most abundant operational taxonomic unit (OTU6782) in all the fungal communities, was isolated from wild cicada flowers. Furthermore, the effects of cicada pupae on metabolite synthesis in Isaria cicadae TZC-3 were studied in submerged culture. The contents of intercellular polysaccharides, adenosine, N6-(2-hydroxyethyl)-adenosine, free amino acids, and hydrolyzed monosaccharides in the mycelia cultured with cicada pupa powder (4%) were significantly increased as compared with the contents in the control group. This indicates that a cicada pupa can act as an elicitor for metabolite synthesis in Isaria cicadae.

Edible larvae and pupae of honey bee (Apis mellifera): Odor and nutritional characterization as a function of diet.

Future protein demand is expected to rise with global population growth. In this study a comprehensive sensorial analysis of the odor of honey bee (Apis mellifera) larvae and pupae as function of their diet (with and without added sugar solution) was performed, as well as nutritional values and antioxidant activity analysis. Honey bee brood powder is a potentially valuable nutritional source with 20-25% protein (dry matter basis), high antioxidant activity and polyphenol content. Main volatile compounds detected using GC-MS with HS-SPME injection were odorless pheromones that represented differences between larvae and pupae. The determined active odor compounds were 2- and 3-methylbutanal, diacetyl, nonanal, dimethyl sulfide and ocimene. A trained sensory panel described honey bee brood aroma profile mainly with buttery and milky attributes, with different life stages and diets giving similar profiles. Such studies can be useful for future development of food products with desired nutritional and sensorial characteristics.

Berberis sawfly contains toxic peptides not only at larval stage.

Livestock can die from grazing in areas where larvae of certain Argidae or Pergidae species containing toxic peptides occur in mass. However, it remains unknown whether other stages also contain these compounds. Here, single specimens of larvae, prepupae, and adults of Arge berberidis, plus samples of its cocoons and larval feces, were analyzed by liquid chromatography-tandem mass spectrometry. The four peptides, pergidin (Perg), 4-valinepergidin (VPerg), dephosphorylated pergidin (dpPerg), and lophyrotomin (LGln), were detected in each of the three stages. Peptide concentrations, in percentage fresh weight, increased from larval up to adult stages, with mean values from 0.044 to 0.125% for Perg, 0.008 to 0.023% for VPerg, and 0.064 to 0.116% for LGln, whereas dpPerg never exceeded 0.001%. The concentrations of this latter peptide averaged 0.002% in the cocoon built by the prepupa, and nearly no peptides were detected in larval feces. Moreover, the concentrations of the three main peptides (Perg, LGln, and VPerg) tended to be correlated with each other in larvae and especially in adults. It is likely that peptide production, purportedly by an endosymbiont, stops at prepupal stage and that concentration of the peptides increases from prepupa to adult due to a decrease of body weight.

Protective effect of silkworm pupa oil on hydrochloric acid/ethanol-induced gastric ulcers.

BACKGROUND: Silkworm pupae are a traditional Chinese food, rich in various saturated and unsaturated fatty acids. Unsaturated fatty acids have a certain protective effect against oxidative damage. The present study used an animal model to determine the protective effect of silkworm pupa oil on hydrochloric acid / ethanol-induced gastric ulcer. RESULTS: Silkworm pupa oil is rich in unsaturated fatty acids, including palmitoleic acid 63.4 g kg-1 , oleic acid 249.1 g kg-1 , linoleic acid 47.0 g kg-1 , and linolenic acid 337.8 g kg-1 , whereas its unsaturated fatty acid content is 700 g kg-1 . Compared to a gastric ulcer control group, high and low doses of pupa oil reduced gastric ulcer area and gastric secretion, whereas gastric pH increased. It also increased serum antioxidant superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) levels, somatostatin (SST), and vasoactive intestinal peptide (VIP) levels, and reduced serum interleukin-6 (IL-6), interleukin-12 (IL-12), tumor necrosis factor (TNF-α), and interferon-γ (IFN-γ), motilin (MTL), and gastrin (GT) levels. RT-qPCR and western blot analyses indicated that silkworm pupa oil significantly increased CAT, GSH-Px, epidermal growth factor (EGF), Epidermal growth factor receptor (EGFR), vascular endothelial growth factor (VEGF), endothelial nitric oxide synthase (eNOS), Cu/Zn-SOD, Mn-SOD, and NF-kappa-B inhibitor-α (IκB-α) expression and lowered nuclear factor-kappa B (NF-κB), B-cell lymphoma-2 (Bcl-2), cyclooxygenase-2 (COX-2), and inducible nitric oxide synthase (iNOS) expression. CONCLUSION: Silkworm pupa oil treatment reduced oxidative damage and inflammation in mice, and high-dose silkworm pupa oil was superior to low-dose silkworm pupa oil, following ranitidine. © 2018 Society of Chemical Industry.

Encapsulation and Systemic Delivery of 5-Fluorouracil Conjugated with Silkworm Pupa Derived Protein Nanoparticles for Experimental Lymphoma Cancer.

Protein-based drug delivery systems have an edge over conventional drug delivery systems due to their biodegradability, non-antigenicity, and excellent biocompatibility to improve the therapeutic properties of anticancer drugs. This study describes the increased anticancer efficacy of 5-fluorouracil (5-FU) conjugated with silkworm Bombyx mori pupal biowaste derived nanoparticles. Here, we have checked the toxicity of pupa-protein nanoparticles (PpNps) and their potential as a carrier for anticancer drugs. PpNps were prepared by a desolvation method which resulted in a uniform particle size of 162.7 ± 2.9 nm. The 5-FU loaded PpNps were formulated and characterized. The drug content of the developed 5-FU conjugated nanoparticles was evaluated by HPLC analysis. The entrapment efficiency and loading capacity of 5-FU were analyzed by HPLC and determined to be 93% and 88.6%, respectively. The release studies showed the biphasic release of 5-FU at pH 7.4 where rapid drug release was achieved for first 30 min, followed by a sustained release of 5-FU from the developed Nps achieved for the next 8 h. Mice with developed ascites tumors were intraperitoneally treated with 5-FU-PpNps and sacrificed. There was a significant increase in total red blood cells and hemoglobulin in 5-FU-PpNps treated mice, whereas a significant decrease in white blood cells which indicated the reduced inflammation of cancer. Subsequently, 5-FU-PpNps decreased the tumor volume and tumor cell viability, which proved its cytotoxic property to cancer cells. This study presents a novel approach to derive B. mori pupal protein nanoparticles, which can be safely used for cancer drug delivery.

Silkworm Pupa Protein Hydrolysate Induces Mitochondria-Dependent Apoptosis and S Phase Cell Cycle Arrest in Human Gastric Cancer SGC-7901 Cells.

Silkworm pupae (Bombyx mori) are a high-protein nutrition source consumed in China since more than 2 thousand years ago. Recent studies revealed that silkworm pupae have therapeutic benefits to treat many diseases. However, the ability of the compounds of silkworm pupae to inhibit tumourigenesis remains to be elucidated. Here, we separated the protein of silkworm pupae and performed alcalase hydrolysis. Silkworm pupa protein hydrolysate (SPPH) can specifically inhibit the proliferation and provoke abnormal morphologic features of human gastric cancer cells SGC-7901 in a dose- and time-dependent manner. Moreover, flow cytometry indicated that SPPH can induce apoptosis and arrest the cell-cycle in S phase. Furthermore, SPPH was shown to provoke accumulation of reactive oxygen species (ROS) and depolarization of mitochondrial membrane potential. Western blotting analysis indicated that SPPH inhibited Bcl-2 expression and promoted Bax expression, and subsequently induced apoptosis-inducing factor and cytochrome C release, which led to the activation of initiator caspase-9 and executioner caspase-3, cleavage of poly (ADP-ribose) polymerase (PARP), eventually caused cell apoptosis. Moreover, SPPH-induced S-phase arrest was mediated by up-regulating the expression of E2F1 and down-regulating those of cyclin E, CDK2 and cyclin A2. Transcriptome sequencing and gene set enrichment analysis (GSEA) also revealed that SPPH treatment could affect gene expression and pathway regulation related to tumourigenesis, apoptosis and cell cycle. In summary, our results suggest that SPPH could specifically suppress cell growth of SGC-7901 through an intrinsic apoptotic pathway, ROS accumulation and cell cycle arrest, and silkworm pupae have a potential to become a source of anticancer agents in the future.

Ultrafast Screening of a Novel, Moderately Hydrophilic Angiotensin-Converting-Enzyme-Inhibitory Peptide, RYL, from Silkworm Pupa Using an Fe-Doped-Silkworm-Excrement-Derived Biocarbon: Waste Conversion by Waste.

A novel, moderately hydrophilic peptide (RYL) with high ACE-inhibitory activity was screened ultrafast via a concept of waste conversion using waste. This novel peptide was screened from silkworm pupa using an Fe-doped porous biocarbon (FL/Z-SE) derived from silkworm excrement. FL/Z-SE possessed magnetic properties and specific selection for peptides due to Fe's dual functions. The selected RYL, which has moderate hydrophilicity (LogP = -0.22), exhibited a comparatively high ACE-inhibitory activity (IC50 = 3.31 ± 0.11 µM). The inhibitory kinetics and docking-simulation results show that, as a competitive ACE inhibitor, RYL formed five hydrogen bonds with the ACE residues in the S1 and S2 pockets. In this work, both the screening carbon material and the selected ACE-inhibitory peptide were derived from agricultural waste (silkworm excrement and pupa), which offers a new way of thinking about the development of advanced uses of the silkworm byproducts and wastes.

Angiotensin-converting enzyme inhibitory and antioxidant peptides from digestion of larvae and pupae of Asian weaver ant, Oecophylla smaragdina, Fabricius.

BACKGROUND: Mixed larvae and pupae of weaver ant (Oecophylla smaragdina) are widely used as an important food ingredient in regions of Thailand. They have high nutritional values and comprise 53% protein and 13% lipid. Peptides derived from food proteins have been shown to possess biological activities. RESULTS: Peptides derived from pepsin and trypsin digestion of these weaver ant larvae and pupae were purified based on angiotensin-converting enzyme (ACE) inhibitory and antioxidant activities, and their amino acid sequences were identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS). In silico docking of peptides with ACE successfully predicted the inhibitory peptides as confirmed by their chemical synthesis. Two peptides with sequences of FFGT and LSRVP showed IC50 values for ACE inhibition of 19.5 ± 1.7 and 52.7 ± 4.0 µmol L-1 , respectively. In addition, one potent antioxidant peptide with a sequence of CTKKHKPNC showed IC50 values of 48.2 ± 2.1 µmol L-1 for DPPH assay and 38.4 ± 0.2 µmol L-1 for ABTS assay, respectively. CONCLUSION: These results indicate that proteins from larvae and pupae of weaver ants are potential sources of peptides with anti-ACE and antioxidation bioactivities. © 2016 Society of Chemical Industry.

Silkworm pupae oil exerts hypercholesterolemic and antioxidant effects in high-cholesterol diet-fed rats.

BACKGROUND: Silkworm pupae is a good resource of edible oil that is especially rich in unsaturated fatty acids and is considered to be an excellent dietary supplement for hyperlipidemia. RESULTS: Groups fed a high-cholesterol diet (HCD) with silkworm pupae oil (SPO) supplementation (1, 2, or 4 mL kg-1 day-1 ) orally had significantly lower levels of serum total cholesterol (P < 0.05) and low-density lipoprotein cholesterol (P < 0.05) compared to the HCD group. With regard to antioxidant parameters, except for levels of glutathione peroxidase (GSH-Px) in the liver, 2 and 4 mL kg-1 day-1 of SPO supplementation leaded to higher total antioxidant capacity (P < 0.05), superoxide dismutase (P < 0.05) and GSH-Px levels (P < 0.05), as well as lower malondialdehyde levels (P < 0.05), both in serum and liver compared to the HCD group. CONCLUSION: The results of the present study indicate that supplementation with SPO can improve lipid profiles and alleviate oxidative stress in high-cholesterol diet-fed rats. © 2016 Society of Chemical Industry.