Optimization of Scorpion Protein Extraction and Characterization of the Proteins' Functional Properties.

Scorpion has long been used in traditional Chinese medicine, because whole scorpion body extract has anti-cancer, analgesic, anti-thrombotic blood anti-coagulation, immune modulating, anti-epileptic, and other functions. The purpose of this study was to find an efficient extraction method and investigate some of physical and chemical parameters, like water solubility, emulsification, foaming properties, and oil-holding capacity of obtained scorpion proteins. Response surface methodology (RSM) was used for the determination of optimal parameters of ultrasonic extraction (UE). Based on single factor experiments, three factors (ultrasonic power (w), liquid/solid (mL/g) ratio, and extraction time (min)) were used for the determination of scorpion proteins (SPs). The order of the effects of the three factors on the protein content and yield were ultrasonic power > extraction time > liquid/solid ratio, and the optimum conditions of extraction proteins were as follows: extraction time = 50.00 min, ultrasonic power = 400.00 w, and liquid/solid ratio = 18.00 mL/g. For the optimal conditions, the protein content of the ultrasonic extraction and yield were 78.94% and 24.80%, respectively. The solubility, emulsification and foaming properties, and water and oil holding capacity of scorpion proteins were investigated. The results of this study suggest that scorpion proteins can be considered as an important ingredient and raw material for the creation of water-soluble supramolecular complexes for drugs.

Biochemical characterization, and anti-inflammatory and antitumor activities of glycoprotein from lamb abomasum.

ETHNOPHARMACOLOGICAL RELEVANCE: Lamb abomasum is used as an edible medicinal source in traditional Chinese medicine for the treatment of gastrointestinal disorders. Lamb abomasum sourced biochemical drug Lamb's trip extract and Vitamin B12 capsule used for the clinical treatment of chronic gastritis, gastric ulcer, and reversal of intestinal metaplasia. Therefore, claimed to have prevention of gastric cancer activity. AIM OF THE STUDY: In this study, we aim to assess whether the glycoprotein has biological activity in the cure of gastric disorder and conduct a structure-activity relationship. MATERIALS AND METHODS: Glycoproteins' extraction conditions were optimized by the response surface method and purified with DEAE-cellulose and Sephadex G-50 chromatography. Two homogenous glycoproteins' physiochemical structures were studied with electrophoresis, HPLC analysis, peroxide oxidation, and ß-elimination, FT-IR, CD, LC-MS/MS, and EDS analysis. The antiinflammation activity of the glycoprotein was determined against COX-2 and LOX-15 enzyme inhibitory ability in vitro, and antitumor activity against HT-29 and HGC-25, and cytotoxicity on L-02 cells was determined in vivo with the MTT method. RESULTS: The abomasum was abundant in glycoprotein and the extraction yield of glycoprotein was up to 24.6 ± 2.1% under optimized conditions. Two homogeneous glycoproteins SAGP-I and SAGP-II determined to be ribose-conjugated and sulfated glycoproteins with a molecular weight of 15.6 kDa and 6.4 kDa. And according to the structural analysis, SAGP-I was a mucin-type ribose-conjugated glycoprotein with 14 O-glycosylation and one N- glycosylation site. SAGP-I and SAGP-II have remarkable anti-inflammatory activity against COX-2 enzyme with the IC50 of 17.64 ± 1.25 µg/mL and 16.14 ± 1.11 µg/mL, respectively. Meanwhile, the two glycoproteins showed strong antitumor activity against HT-29 with the EC50 of 19.19 ± 1.46 µg/mL and 184.9 ± 5.6 µg/mL, respectively. CONCLUSION: The Highly purified glycoprotein SAGP-1 and SAGP-II showed anti-inflammatory activity against the COX-2 enzyme, and antitumor activity against HT-29 human colon cancer cells and noun-inhibitory activity against LOX-15 enzyme and HGC-25. Both glycoproteins are ribose conjugated and sulfated whose characters are related to their anti-inflammatory and anti-tumor activity. Such results suggest the possibility of anti-inflammatory and pre-cancer activity. And in some degree explains the pharmacy of abomasum's traditional use in gastric disorder and clinical use of lamb abomasum APIs drugs' in gastric disorders and gastric cancer development. This study provides a preliminary basis for the further study of the per-cancer mechanism of lamb abomasum glycoprotein. And, would be the material basis of the clinical use of Lamb's trip extract and Vitamin B12 capsule.

Covalent binding of flavonoids with silk sericin hydrolysate: Anti-inflammatory, antioxidant, and physicochemical properties of flavonoid-sericin hydrolysate conjugates.

To construct food ingredients with improved bioactivities and physicochemical properties, two sericin hydrolysate-flavonoid conjugates, bearing quercetin and rutin covalently bound to sericin, were prepared under alkaline conditions. UV spectroscopy and SDS-PAGE confirmed that sericin hydrolysate and oxidized flavonoids formed conjugates, which were primarily the result of covalent interactions. Changes and differences in molecular weight distribution, secondary and tertiary structures, surface hydrophobicity, and surface morphology were observed. Anti-inflammatory activities were evaluated by basing on inhibitory activity against nitric oxide (NO) production and 15-lipoxygenase (15-LOX). The conjugates showed significantly improved (p < .05) anti-inflammatory and emulsifying properties than unmodified sericin hydrolysate. Meanwhile, the covalent interaction had a positive effect on the antioxidant activity of sericin hydrolysate. PRACTICAL APPLICATIONS: We had prepared flavonoid-sericin hydrolysate conjugates and evaluated the effect of covalent binding of flavonoids on the physicochemical, anti-inflammatory, antioxidant, and emulsifying properties of sericin hydrolysate, which is beneficial to broaden the range of applications of sericin and flavonoids in the food, cosmetics, and pharmaceutical industries.

Effects of trypsin-induced limited hydrolysis on the structural, functional, and bioactive properties of sericin.

The effects of trypsin-induced hydrolysis on the structural, functional, and antioxidant properties of sericin were studied. The structural properties of sericin and its hydrolysates were characterized by using SDS-PAGE, SEC-HPLC, surface hydrophobicity, and circular dichroism. Antioxidative properties were evaluated based on quenching capacity against hydroxyl, DPPH, and ABTS, and metal (Fe2+, Cu2+) chelating activity. The enzymatic hydrolysis raised the flexibility, changed emulsifying and foaming properties, and improved the solubility and antioxidant activity of sericin. Meanwhile, the hydrolysis led to a decline in gelation capacity, oil holding capacity, and water holding capacity. Sericin and its hydrolysates exhibited excellent function with regard to oil holding, emulsifying, and foaming. Sericin and its hydrolysates had clear effects on the growth of both Enterococcus faecalis and Lactobacillus bulgaricus strains.

Study on the bioactive ingredients of lamb abomasum medicine / 中华消化杂志

Objective:To study the material basis of the drug effect of the raw material of lamb′s tripe and vitamin B12 capsule.Methods:Rennet and pepsin were extracted with 0.9% sodium chloride and were purified by saturated ammonium sulfate precipitation, diethylaminoethyl-cellulose 52 and high pressure chromatography (HPLC) chromatography. Relative molecular weight was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) electrophoresis and liquid chromatography-triple quadrupole mass spectrometer. The amino acid composition and antioxidant activity of raw materials were analyzed with HPLC. The raw materials were completely extracted with water, phosphate buffer and sodium bicarbonate in turn, and in vitro 1, 1-diphenyl-2-picrylhydrazyl radil 2, 2-diphenyl-1-(2, 4, 6-trinitrophenyl)hydrazyl (DPPH) and 2, 2′-azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS) radical activity were measured. Glycoprotein from raw material was extracted with hot water and determined its growth-promoting activity gaginst Bifidobacterium adolescentis , Lactobacillus delbrueckii subsp. Bulgaricus and Enterococcus faecalis was measured. The composition of amino acid and monosaccharide were analyzed by HPLC and gas chromatography-mass spectrometry, respectively. Results:The enzyme activity of two purified rennet F6-2 and F7-2 which had different ionic strengths and pepsin F7-1 were 27 557.10, 17 532.60 and 17 728.15 U/g, respectively. The results of SDS-PAGE indicated that the molecular weights of three enzymes were similar, ranging from 35 000 to 40 000. The raw material contained 16 kinds of amino acids, of which hydrophobic amino acids accounted for 33.03% of the total amino acid content. When the sample concentration was 5 mg/mL, ABTS free radical scavenging activity of the three extracts was (37.80±0.45)%, (23.20±0.78)% and (62.80±0.74)%; DPPH free radical scavenging activity was (57.87±0.55)%, (5.03±0.25)% and (26.67±3.10)%, respectively. Glycoprotein extracts had promoted the growth of Bifidobacterium adolescentis, Lactobacillus delbrueckii subsp. Bulgaricus and Enterococcus faecalis, and there was statistically significant difference in the promotion of Lactobacillus delbrueckii subsp. Bulgaricus and Enterococcus faecalis ( P<0.05). The protein chain of glycoprotein was composed of 15 amino acids and the polysaccharide chain was composed of two monosaccharides, glucose and lactose. Conclusions:Two rennet and one pepsin are isolated and analyzed from raw material of lamb abomasum by various chromatographic methods. The raw material is rich in antioxidant active ingredients. The glycoprotein components of lamb abomasum has the activity of promoting the growth of probiotics.