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Lectins from plants of the Diocleinae subtribe often exhibit specificity towards mannose/glucose and derived sugars, with some plants also displaying a second lectin specific to lactose/GalNAc. Here, we present a novel lectin from Collaea speciosa, named CsL, that displays specificity for GlcNAc/glucose. The lectin was extracted from Collaea speciosa seeds and purified by a single chromatographic step on a Sephadex G-50 matrix. In solution, the lectin appears as a dimeric protein composed of 25 kDa monomers. The protein is stable at pH 7–8 and dependent on divalent cations. CsL maintained its agglutination activity after heating to 90 °C for 1 h. Glycan array studies revealed that CsL binds to N-glycans with terminal GlcNAc residues, chitobiose and chitotriose moieties. The partial amino acid sequence of the lectin is similar to that of some lactose-specific lectins from the same subtribe. In contrast to other ConA-like lectins, CsL is not toxic to Artemia. Because of its remarkably different properties and specificity, this lectin could be the first member of a new group inside the Diocleinae lectins.

This study investigated the structural elucidation, anti-radical and immunomodulatory activities of polysaccharides from the roots of Glehnia littoralis. A crude polysaccharide was extracted from the roots of G. littoralis through the ultrasonic-assisted extraction and further purified by DEAE-52 cellulose and Sephadex G-100 gel column, a major polysaccharide fraction named GLP80-1 was obtained. The chemical properties and structure of GLP and GLP80-1 were characterized by acid hydrolysis, methylation analysis, along with high performance gel permeation chromatography, Fourier transform infrared, nuclear magnetic resonance, scanning electron microscope, thermogravimetric analysis and X-ray diffraction. The molecular weight distributions of GLP were determined as 1.89 × 106 and 1.26 × 104 Da. The monosaccharide composition of GLP was glucose, glucuronic acid, galactose and arabinose with molar ratios of 0.91:0.04:0.03:0.02, respectively. The average molecular weight of GLP80-1 was determined as 1.63 × 104 Da. The structure of GLP80-1 was deduced to be a homogenous glucan, comprised a main chain of (1→4)-linked-α-D-Glcp with a single α-D-Glcp branch substituted at C-6. The results of biological activities in vitro showed that GLP and GLP80-1 exhibited free radical scavenging effects, and displayed promotion for the proliferation of mouse spleen lymphocytes and RAW264.7 cells. The data indicated that GLP and GLP80-1 had the potential to be explored as novel natural antioxidant and immunomodulator for application in functional food.

This study investigated the phytochemical constituents and evaluated the total phenolic content, it also determined the antioxidant and antibacterial activities of the ethanol (EtOH) extract of the whole plant of Dicliptera verticillata. The phytochemical study was carried out using repeated silica gel column chromatography followed by Sephadex LH-20 to purify the extract which afforded three known compounds: lutein (1), shikimic acid (2) and guaiacylglycerol (3). The structures of these compounds were identified after analyses of their NMR and MS data and comparison of these data with those reported in the literature. The antioxidant and antibacterial activities were evaluated by the 2,2-diphenyl-1-picrylhydrazyl radical assay and the broth macrodilution method, respectively. The EtOH extract showed moderate antioxidant activity (63.9%; IC50 40.16 μg/mL), weak total phenolic content and had a bactericidal effect against both Gram-positive (Staphylococcus aureus) and Gram-negative bacteria (Klebsiella pneumonia and Escherichia coli). This is the first report on the isolation of compounds (1–3), as well as the total phenolic content and the antibacterial and antioxidant activities of the EtOH extract of D. verticillata.

The polysaccharide composition of dried bulbs of Orchis mascula (L.) L. is reported. The obtained polysaccharides were studied as sulfate derivatives, were called SP-S, and were purified by ion-exchange chromatography over a column of DEAE 650M to afford four fractions. Homogeneous polysaccharides were obtained by purifying these fractions using a column of Sephadex G-150. The chemical characteristics were studied. The antioxidant properties of these fractions were determined.

The antidiarrhoeal effect of Waltheria indica methanol extract and fractions have been reported earlier but, the present work examined the intestinal relaxant effects of two flavonoid-phenyl propanoids isolated from the methanol extract. The active aqueous fraction was subjected to vacuum liquid chromatography using dichloromethane with increasing concentration of ethyl acetate, and that of methanol and water successively. The ten (10) fractions obtained were combined to give seven (7). The fraction 2 (C, D) was subjected to preparative thin layer chromatography on silica gel GF254 (10-40μm) using CHCl3-CH3OH (8:2) to obtain compound coded F2. Fraction 4 (F) was subjected to column chromatography using silica gel (60-120μm mesh) and eluted with dichloromethane with increasing concentrations of methanol. Fractions 9-28 were combined and subjected to column chromatography using chloroform with increasing concentration of methanol. The fractions 1-16 of these were purified on Sephadex LH-20 to obtain compound BAA. The identities of the two compounds were established using spectroscopic methods. The antidiarrheal effect of compound F2 was evaluated on mice using charcoal transit (100,200, 400mg/kg), castor oil (40, 60 mg/kg) while the two compounds were examined for their inhibitory effects on Ach-induced ileum contraction. The effects of the compounds were compared with loperamide (3mg/kg) and atropine (80μg). Compounds F2 and BAA were identified as tiliroside and 3’’’, 5’’’-dimethoxy tiliroside respectively. Tiliroside inhibited the charcoal transition in the animals in a dose dependent pattern with 400mg/ mL eliciting 63.41% inhibition compared to 59.23% produced by loperamide. The compound also elicited significantly (P

Purpose: This study sought to find out the knowledge/awareness level of men on birth preparedness and complication readiness in Magarini Sub County. Methodology: A cross-sectional study sequential mixed methods design was used where a total of 464 men will be enrolled. Quantitative data was collected using semi structured questionnaires and interview guides were used to collect qualitative data. Quantitative data was coded, and analyzed by SPSS software. Qualitative data was analyzed using NVIVO software. Chi- square test was used to determine associations between categorical variables and Logistics regression was used to identify factors associated with birth preparedness and complication readiness. The associations between awareness and each independent variable were determined by odds ratio (OR) and 95% confidence interval (CI). Thematic content analysis was applied for qualitative data analysis. Findings: The result indicated that the odds of pregnancies resulting in a baby that was born alive were 47.306 times higher for more than two pregnancies as compared to one pregnancy(Odds=47.306,p=0.000). The odds of pregnancies resulting in a baby that was born alive were 16.25 times higher for one pregnancies as compared to no pregnancy(Odds=16.25,p=0.000). Unique contribution to theory, practice and policy Birth Preparedness and Complication Readiness (BPACR) should be endorsed as an essential component of safe motherhood programs to reduce delays for care-seeking for obstetric emergencies and this has been proven to positively impact on birth outcomes Keywords: knowledge/awareness level, birth preparedness, complication readiness, Magarini Sub County

Xylanases (EC 3.2.1.8) have been considered as a potential green solution for the sustainable development of a wide range of industries including pulp and paper, food and beverages, animal feed, pharmaceuticals, and biofuels because they are the key enzymes that degrade the xylosidic linkages of xylan, the major component of the second most abundant raw material worldwide. Therefore, there is a critical need for the industrialized xylanases which must have high specific activity, be tolerant to organic solvent or detergent and be active during a wide range of conditions, such as high temperature and pH. In this study, an extracellular xylanase was purified from the culture broth of Aspergillus niger VTCC 017 for primary structure determination and properties characterization. The successive steps of purification comprised centrifugation, Sephadex G-100 filtration, and DEAE-Sephadex chromatography. The purified xylanase (specific activity reached 6596.79 UI/mg protein) was a monomer with a molecular weight of 37 kDa estimating from SDS electrophoresis. The results of LC/MS suggested that the purified protein is indeed an endo-1,4-β-d-xylanase. The purified xylanase showed the optimal temperature of 55 °C, and pH 6.5 with a stable xylanolytic activity within the temperature range of 45–50 °C, and within the pH range of 5.0–8.0. Most divalent metal cations including Zn2+, Fe2+, Mg2+, Cu2+, Mn2+ showed some inhibition of xylanase activity while the monovalent metal cations such as K+ and Ag+ exhibited slight stimulating effects on the enzyme activity. The introduction of 10–30% different organic solvents (n-butanol, acetone, isopropanol) and several detergents (Triton X-100, Tween 20, and SDS) slightly reduced the enzyme activity. Moreover, the purified xylanase seemed to be tolerant to methanol and ethanol and was even stimulated by Tween 80. Overall, with these distinctive properties, the putative xylanase could be a successful candidate for numerous industrial uses.

Asparaginases important role in the treatment of leukemia. It is part of chemotherapy in the treatment of leukemia in the last three decades. l-Asparaginase is isolated from Fusarium sp. isolated from soil and purified using ammonium sulfate precipitation and Sephadex G 100. Characterization of the crude enzyme revealed it is a metalloprotease inhibited by EDTA. Hg2+, Cd2+, and Pb2+ also inhibited the enzyme. Mg2+, Zn2+, and Ca2+ activated l-asparaginase. Furthermore, kinetic studies of purified enzyme were carried out. Vmax and Km were 0.031 M and 454 U/mL, respectively. The optimum temperature was 30 °C and the optimum pH was 7. Concerning substrate specificity, gelatin and casein in addition to l-asparagine were tested. The enzyme was found to be nonspecific that could hydrolyze all tested substrates at different rates. The maximum enzyme activity was recorded in the case of l-asparagine, followed by casein and gelatin, respectively. The molecular weight of l-asparaginase was 22.5 kDa. The antileukemic cytotoxicity assay of the enzyme against RAW2674 leukemic cell lines by MTT viability test was estimated. The enzyme exhibited antileukemic activity with IC50 of 50.1 UmL−1. The current work presents additional information regarding the purification and characterization of the enzyme produced by Fusarium sp. and its evaluation as a potential antileukemic chemotherapeutic agent.

This study investigated the larvicidal activity on Culex quinquefasciatus of lectin purified from fresh fruiting bodies of woodland mushroom, Agaricus semotus. A. semotus lectin (ASL) was purified via ion-exchange chromatography on DEAE-cellulose A-25 and size exclusion chromatography on Sephadex G-100 matrix. Molecular weight (16.6 kDa) was estimated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The effects of temperature, pH, metal chelation- and larvicidal activity of ASL were also investigated. The ASL indifferently agglutinated the erythrocytes of the human ABO blood system and was stable at acidic pH and below 50 °C whereas 66% of its activity was lost at 60 °C with complete inactivation at 70 °C. ASL is a metalloprotein requiring barium ion as chelation of metals by 50 mM EDTA rendered the lectin inactive, while the addition of BaCl2, among other metal salts, restored the activity. ASL showed larvicidal activity against C. quinquefasciatus larvae after 24 h with a mortality of 5 and 95% at 5 and 25 mg/mL respectively, and LC50 of 13.80 mg/mL. This study concluded that purified A. semotus lectin showed impressive larvicidal activity, which could be exploited in its development as an insecticidal agent.

Crude polysaccharides from Spirulina platensis (SP) were isolated by maceration with a hot alkali solution and further fractionated by DEAE-52 cellulose and Sephadex G-100 chromatography into two purified fractions PSP-1 and PSP-2. The monosaccharide composition analysis indicated that SP was mainly composed of rhamnose and glucose, while PSP-1 and PSP-2 were composed only of glucose. The composition analysis of PSP-1 and PSP-2 by HPLC, FT-IR, and NMR showed that PSP-1 and PSP-2 were branching dextran, and their structures were (1 → 4)-linked-α-D-Glcp as the main chain, and C-6 replaced the single α-D-Glcp as the linear structure of the branch chain. The glucans (SP/PSP-1/PSP-2) can significantly improve the phagocytic ability of macrophages, enhance iNOS activity, promote NO production, and increase IL-6 mRNA expression, so they may possess certain immunomodulatory activity.

Four neutral polysaccharides (ESBP1-1, ESBP1-2, ESBP2-1 and ESBP3-1) were successfully purified from the water extracted crude polysaccharides of Erythronium sibiricum bulbs through the combination of DEAE Sepharose CL-6B and Sephadex G-100 chromatography; their average molecular weights were 1.3 × 104, 1.7 × 104, 9.4 × 105 and 4.1 × 105 Da, respectively. Monosaccharide component analysis indicated that ESBP1-1 and ESBP1-2 were mainly composed of glucose (Glc). ESBP2-1 was composed of Glc, galactose (Gal) and arabinose, with a molar ratio of 24.3:1.1:1, whereas ESBP3-1 comprised Glc and Gal at a molar ratio of 14.8:1. In-vitro study showed that all of the four polysaccharides were able to considerably promote the proliferation and neutral red phagocytosis of RAW 264.7 macrophage cell. They could also stimulate the production of the cell lines’ secretory molecules [nitric oxide, tumour necrosis factor-α (TNF-α) and interleukin-1β (IL-1β)] in a dose-dependent manner. However, ESBP1-2 was not included in IL-1β. Overall, these results suggested that polysaccharides from E. sibiricum bulbs can be developed as immunomodulatory ingredients for complementary medicines or functional foods. However, further animal or clinical studies are required.

Lipase enzyme plays a part in the environmental and pharmaceutical industries, microbial lipase is the most potential source for industrial application. Lipases were extracted from endogenous isolate, sequentially purified by ammonium sulphate and Sephadex G-150 methods, and immobilized on silver-alginate nanoparticles to test the ability to used preparing predigested of high-fat meat to low fat meat. Lipase producing bacteria was identified as Lactobacillus fermentum, that extracted mesophilic lipases presented maximal activity at pH 8 and 40 °C. The purified enzyme was recovered up to 16% yield in 1.4 fold purification and specific activity 4 U/mg. Lipase activity exhibited the highest inhibition rate by adding 1% of non-ionic detergent of Triton X-100, Tween 80 and Tween 20 (60, 63 and 71%, respectively). Silver-alginate nanoparticles is a potential matrix for immobilization of biomolecules, especially enzymes. Immobilized lipase was preserved; the specific activity (2.3–2 U/mg) can even be reused more than three times, which attributed to hydrophobicity and flexibility of lipases. Predigested meat was obtained from treating high-fat meat with immobilized enzymes, as shown by losses of 40% of initial meat weight after 14 days of lipid digestion. Conjugation of silver-alginate matrix with lipase can controlled the digestion of meat fat and easy to get rid of in comparing to expressed enzyme. Concept of predigested meat could revolutionize the world of high-fat diets without interfering their dietary habits.

This study aimed to isolate and characterize the structures of glycoproteins from peas and determine their hypoglycemic activity. The crude pea glycoproteins (PGP) were extracted by hot water and purified by diethylaminoethyl (DEAE)-Sepharose chromatography and Sephadex G-100 size-exclusion chromatography in sequence. Then three main fractions were obtained, namely PGP1, PGP2 and PGP3, with molecular weights of 897 615, 846 740 and 1 194 692 Da, respectively. The physical and chemical properties of the three fractions were evaluated and compared by Fourier transform infrared spectroscopy (FT-IR), nuclear magnetic resonance (NMR), scanning electron microscope (SEM), high performance liquid chromatography (HPLC) and other analytical techniques. The fraction PGP2 with the highest hypoglycemic activity, was screened using the Caco-2 monolayer cell model. It can inhibit the uptake of glucose in the small intestine, as well as the activities of maltase and sucrase. After simulated gastrointestinal digestion, PGP2 significantly enhanced the inhibitory effect of α-glucosidase, and slightly reduced the inhibitory ability of α-amylase. In summary, PGP2 possessed strong hypoglycemic activity after digestion. These results indicated that PGP2 has the potential to be developed into a functional food or natural medicine for the treatment of type 2 diabetes mellitus.

In this paper, a novel polysaccharide extracted from Hericium erinaceus coded HEP was successfully isolated and purified by DEAE-Sepharose CL-6B and Sephadex G-200 column chromatography, which was applied to study the preparation process of HEP–zinc(II) chelate (HEP–Zn). Chelation rate was used as the index, effect of HEP concentration, reaction temperature and reaction time on chelation rate was investigated. The preparation process of HEP–Zn was optimized by response surface methodology based on the single factor experiments. In addition, antioxidant activities and structural characteristics of HEP and HEP–Zn were compared. The results showed that the optimal preparation process of HEP–Zn was obtained as following: HEP concentration was 15.3 mg/mL, chelating time was 3.0 h and chelating temperature was 40 ℃. Under the optimal preparation process conditions, chelation rate of HEP–Zn was 5.20%. Compared with HEP, HEP–Zn displayed irregular spongy structure that provided large surface area for chelation and processed better DPPH radical scavenging activity and stronger superoxide anion scavenging activity. The results indicated that HEP–Zn had enormous potential to be used as a novel zinc supplement or antioxidant in food industry.

The walnut protein hydrolysate (WPH) was prepared via simulated gastrointestinal digestion. The degree of hydrolysis (DH), amino acid composition, and relative molecular weight distribution of WPH were analyzed. The results showed that the DH of WPH was 11.6%, WPH was rich in Glu and Pro, and the relative average molecular weight of 572 Da accounted for 59.78%. The effects of WPH on osteoporosis were evaluated using a model of retinoic acid-induced osteoporosis rat. Treatment with WPH effectively increased the serum calcium and phosphorus contents, alleviated calcium loss, and reduced tartrate-resistant acid phosphate and alkaline phosphatase activities and bone gla protein content. WPH treatment significantly improved the biomechanical properties of the bone and increased the value of bone mineral density. In addition, WPH treatment improved the bone microstructure. WPH was isolated and purified by Sephadex G-25 gel filtration chromatography and semi-preparative reversed-phase high-performance liquid chromatography. A fraction with high calcium-binding activity was obtained and 15 peptides were identified.

A novel acidic polysaccharide, named as AWPA, was extracted form Annona squamosa residue by 0.1 M NaOH alkaline solution and purified by DEAE-cellulose and Sephadex G-150. HPLC analysis indicated that AWPA was a homogeneous polysaccharide with molecular weight of 3.08 × 103 kDa. The monosaccharide composition of AWPA, determined by ion chromatography, was consisted of L-arabinose, D-galactose, d -glucose, D-mannose, D-galacturonic acid in a percentage of 15.58:13.48:60.14:9.02:1.78, respectively. The results of FT-IR, methylation and NMR showed that the sugar residue of AWPA were mainly composed of α-L-Araf-(1→, →4)-α-D-Glcp-(1→, →4)-β-D-Galp-(1→, →6)-β-D-Glcp-(1→, →4,6)-β-D-Galp(1→, →3,6)-α-D-Manp-(1→, respectively. The Congo red experiment on AWPA showed that there was helix conformation. The microstructure of AWPA was detected by scanning electron microscopy, showing that the shape of AWPA was reticular and its structure was irregular. AWPA had effectively α-glucosidase inhibitory activity and α-amylase inhibitory activity with IC50 of 0.667 mg/mL and 1.360 mg/mL, respectively. The inhibitory effects of AWPA on α-glucosidase and α-amylase were both reversible with mixed type and competitive type competition, respectively. The significance of manuscript was not only to avoid the waste of Annona squamosa residue, but provided alternative in the developments of inhibitors of α-glucosidase and α-amylase.

The extracellular polysaccharide of Morchella esculenta cultivated under submerged fermentation was extracted. A single polysaccharide was purified through DEAE-Cellulose 52 and Sephadex G 100, and named as MEP 2a. The molecular weight of MEP 2a was determined by HPGPC and it is about 1391.5 kDa. MEP 2a is composed of mannose and glucose as the monosaccharide unit with a molar ratio of 8.15 : 1.07. The main polysaccharide chemical structure was analyzed by 1D and 2D NMR. Methylation and NMR analysis revealed that the backbone of MEP 2a consists of 1,3,4-linked-Manp, 1,2-linked-Manp and 1,6-linked-Glcp. 1D and 2D NMR results indicated that the main chain is based on →1)-β-D-Glcp-(6→, →1)-α-D-Manp-(3,4→, →1)-α-D-Manp-(2→) and the branch chain is composed of α-D-Manp-(1→, →1)-β-D-Glcp-(6→ and α-D-Glcp-(1→). MEP 2a promoted the phagocytosis function and secretion of NO, IL-1β, IL-6 and TNF-α of macrophages. In the present study, the chemical structure and immunomodulatory ability of an extracellular polysaccharide of Morchella esculenta was investigated which guarantees further research studies and promising applications.

One isoform of NADP+-dependent decarboxylating malate dehydrogenase (EC 1.1.1.40) was found in the leaf mesophyll of corn. The enzyme was purified in four stages: homogenization, fractionation with ammonium sulfate, gel filtration on Sephadex G-25, and ion exchange on Sephacel. The specific activity of the purified, electrophoretically homogeneous preparation was 92 E/mg of protein, while its yield was 15%, and the purification degree was 109. The relative electrophoretic mobility of the decarboxylating malate dehydrogenase was 0.1. The optimal pH for enzyme functioning during the catalysis of forward and reverse reactions was 8.0. The Michaelis constants were determined for the direct and reverse reactions: 5.5 mM for malate and 1.3 mM for pyruvate. Moreover, the affinity for coenzymes varied significantly. Thus, KM was equal to 4.5 mM for NADP+ and 0.9 mM for NADP.

One hundred samples from various clinical sources were obtained. Sixty isolates of Staphylococcus aureus and one isolate of Staphylococcus gallinarum have been identified. The Capability of S. gallinarum was phenotypically tested on DNase agar medium and also by quantitative assay which showed that S. gallinarum has been able to generate the thermostable DNase. DNase was extracted, the activity and specific activity of the crude was 36 (U/ml) and 240(U/mg) respectively. The purification of the enzyme was done by precipitation with ammonium sulphate at saturation (65–85%) then by using ion exchange chromatography in CM cellulose and gel filtration by in Sephadex G150.The activity and specific activity Purified DNase was 35 (U/ml) and 3500(U/mg) respectively. The optimal PH for DNase was found to be 8 while the enzyme was stable at broad pH range (7, 8, 9 and 10) with remaining activity 94%, 100%, 97%, 86% respectively. The optimal temperature for DNase activity and stability was at 37 °C. The results indicate that DNase activity increased when 10 mM of each MnCl2, KCl, NaCl MgCl2 and CaCl2 were incubated with the enzyme. The molecular weight of DNase was estimated by gel filtration and found that it was approximately 27 KDa.

Egletes viscosa is a plant with therapeutic value due to its antibacterial, antinociceptive and gastroprotective properties. This study aimed to purify, characterize, and evaluate the cytotoxicity of a lectin (EgviL) from the floral capitula of E. viscosa. The lectin was isolated from saline extract through precipitation with ammonium sulfate followed by Sephadex G-75 chromatography. The molecular mass and isoelectric point (pI) of EgviL were determined as well as its temperature and pH stability. Physical–chemical parameters of interaction between EgviL and carbohydrates were investigated by fluorescence quenching and 1H nuclear magnetic resonance (NMR). Cytotoxicity was investigated against human peripheral blood mononuclear cells (PBMCs) and neoplastic cells. EgviL (28.8 kDa, pI 5.4) showed hemagglutinating activity stable towards heating until 60 °C and at the pH range 5.0–7.0. This lectin is able to interact through hydrophobic and electrostatic bonds with galactose and glucose, respectively. EgviL reduced the viability of PBMCs only at the highest concentration tested (100 μg/mL) while was toxic to Jurkat E6–1 cells with IC50 of 24.1 μg/mL,inducing apoptosis. In summary, EgviL is a galactose/glucose-binding protein with acidic character, stable to heating and with cytotoxic effect on leukemic cells.

Encapsulation is a vital cellular immune reaction of host insects against endoparasitoids; however, how encapsulation is regulated is still unclear. Utilizing a cell line, SYSU-OfHem C, derived from larval hemocytes of the Asian corn borer Ostrinia furnacalis to assay for encapsulation response, an encapsulation-promoting protein (OfEPP1) was isolated from the plasma of O. furnacalis larvae. OfEPP1 is a novel secretory protein, which exists only in O. furnacalis to date. The OfEpp1 gene is intronless and encodes a protein containing several groups of short repetitive sequences and a high proportion of proline residues (18.3%). OfEPP1 is a thermally stable protein that is mainly expressed in fat bodies, and its accumulation could be induced by the injection of foreign objects (Sephadex beads). Eukaryotically expressed recombinant OfEPP1 promoted hemocytes to encapsulate Sephadex beads, while prokaryotically expressed protein did not, indicating that posttranscriptional modification affects the function of OfEPP1. The encapsulation-promoting function of OfEPP1 could be neutralized by the addition of polyclonal antibodies against OfEPP1 or disrupted by the injection of dsRNA targeting OfEpp1. Eukaryotically expressed OfEPP1 promoted the aggregation, but not spreading, of both granulocytes and plasmatocytes. Immunocytochemistry analysis showed that eukaryotically expressed OfEPP1 could bind to the surface of hemocytes. Therefore, we speculate that OfEPP1 possibly promotes hemocytic encapsulation by binding to the surface of hemocytes as a ligand to induce their aggregation. This study provides evidence clarifying the mechanism of encapsulation in insects.

Ruellia tuberosa L. (RTL) has been used as a folk medicine to cure diabetes in Asia. RTL was previously reported to alleviate hyperglycemia, insulin resistance (IR), abnormal hepatic detoxification, and liver steatosis. However, the potential bioactive compounds of RTL have still not been identified. The aim of this study was to investigate the bioactive compounds in RTL ethyl acetate (EA) fractions by using a glucose uptake assay in TNF-α-treated mouse FL83B hepatocytes to discover a mechanism by which to improve IR. The bioactive compounds were identified by the high-performance liquid chromatography (HPLC) assay. Using the Sephadex LH20 gel packing chromatography column, the EAF5 fraction was isolated from RTL and significantly increased glucose uptake in TNF-α-treated FL83B cells. Moreover, the MCI gel packing chromatography column separated EAF5 into five subfractions and had no significant cytotoxic effect in FL83B cells when treated at the concentration of 25 μg/ml. Among the subfractions, EAF5-5 markedly enhanced glucose uptake in TNF-α-treated FL83B cells. The possible bioactive compounds of the EAF5-5 fraction that were identified by the HPLC assay include syringic acid, p-coumaric acid, and cirsimaritin. The bioactive compound with the best effect of increasing glucose uptake was p-coumaric acid, but its effect alone was not as good as the combined effect of all three compounds of the EAF5-5 fraction. Thus, we speculate that the antidiabetic effect of RTL may be the result of multiple active ingredients.

Probioceuticals are probiotic-derived biologically active compounds that positively influence human health. Exopolysaccharides (EPS) are long chain polymers of sugars and their derivatives with diverse biological functions. The objective of this work was to study EPS from Pediococcus acidilactici (PA) NCDC 252, a potential probiotic. In silico analysis of PA NCDC 252 genome revealed an EPS gene cluster (10 genes) and genes were further analysed for their functional domains and products. EPS from PA NCDC 252 were purified by ethanol precipitation and gel filtration chromatography on Sephadex G-100. Molecular mass of purified EPS was 89.1 KDa as determined by gel filtration chromatography. Structural and component analysis by FTIR, NMR and UPLC revealed purified EPS to be linear homopolysaccharides (α-glucans) with few α-(1→3) branches. in vitro studies showed anti-oxidant, reduction potential and anti-cancer activity in dose dependent manner. Total antioxidant potential of EPS was 11.9 %. In-vitro cell viability assay revealed anti-proliferative effect of EPS on human colon cancer cell line (HCT116). At 10 and 100 μg/mL, EPS inhibited HCT116 cells up to 67.1 % and 87.3 % respectively. These results suggest that PA EPS can be therapeutically used as anti-oxidant and anticancer agent after in vivo studies.

Lectins are a specialized group of proteins with immense biological properties and applications. This study describes the purification and characterization of a lectin from Leucaena leucocephala seeds, a plant belonging to the Fabaceae family. Leucaena leucocephala lectin (LLL) was purified by a two-step purification method involving DEAE-cellulose anion exchange chromatography and Sephadex G-75 size exclusion chromatography. The isolated lectin displayed a high haemagglutination titre upon treatment with rabbit erythrocytes. SDS-PAGE and Reverse-Phase High performance liquid chromatography (RP-HPLC) analysis experimentally revealed the presence of three bands corresponding to 37, 27 and 20 kDa indicating the presence of isolectins. Periodic Acid Schiff's (PAS) staining of LLL confirmed the presence of glycoprotein. Various biochemical parameters were analysed to study its effect on the haemagglutination activity. Sugar inhibition studies experimentally revealed that Glucose was the most potent inhibitor. Fluorescence spectrometric analysis of LLL and Glucose indicated a strong interaction with an association constant of 0.159 × 103 M−1. Circular Dichroism spectroscopy indicated a higher alpha helical content (25.27%). LLL was observed to possess mitogenic activity against Peripheral blood mononuclear cells (PBMC). The present investigation reports the isolation of a novel lectin from this plant which could contribute towards the diagnostic studies of certain diseases and for its therapeutic potential.

This study was aimed to purified and characterized the bacteriocin produced from Lactobacillus acidophilus HT1, in order to use it in a skin pharmaceutical formula. The optimal conditions for bacteriocin production was investigated and results showed that modified nutrient broth was the best medium with glucose (30 gm/L) and yeast extract (7 gm/L) with peptone (7 gm/L) were the optimum carbon and nitrogen sources. In addition, 2% inoculum size, 37C◦ and pH 6.4 were the optimal conditions to obtain maximum bacteriocin of 640 AU/ml after 24 hrs. The bacteriocin was purified using 70% ammonium salt saturation and gel filtration with sephadex G50 that resulted 20 % yield and 2560 AU/ml of activity, then the partial purified bacteriocin was characterized and found the bacteriocin was protein in nature and kept its activity after 10 min at 20, 30 and 40◦C, however 50% of the activity was lost at 50C◦. Moreover, it showed stability at pH 6 and 7 for 30 min whereas; no activity was observed at pH 4 and 9. In addition, results showed that bacteriocin has a bactericidal effect rather than bacteriostatic. A cream formula contained the bacteriocin was prepared which already examined in vitro and in vivo. The effectiveness of the formula was confirmed using Klebsiella sp., Staphylococcus aureus and Pseudomonas aeruginosa as indicator strains. Results established that treatment at the onset time was more effective and the time of healing was decreased.

Firstly, optimal parameters of crude polysaccharide from Buddleja officinalis were obtained as follows: ratio of water to raw material of 26 : 1, ultrasonic power of 240 W, ultrasonic time of 45 min, and ultrasonic temperature of 62°C. Secondly, acidic polysaccharide (APBOM) from Buddleja officinalis was successfully acquired with the yield of 9.57% by using DEAE-52 cellulose and Sephadex G-100 gel column chromatography. Then, we found that total polysaccharide content of APBOM was 94.37% with a sulfuric acid group of 1.68%, uronic acid content of 17.41%, and average molecular weight of 165.4 kDa. Finally, APBOM was confirmed to have significant antiangiogenic effects.

Paenibacillus bovis sp. nov BD3526, isolated from raw yak (Bos grunniens) milk, was able to produce antibacterial substances against Micrococcus luteus. The antibacterial substances produced by the strain BD3526 in 3% (w/v) wheat bran broth under aerobic conditions were precipitated from the cultivated broth with ammonium sulfate at 60% saturation. Two antibacterial compounds were obtained by Sephadex LH-20 chromatography and semi-preparative reverse-phase high-performance liquid chromatography (Semi-Pre RP-HPLC). The chemical structures of the two antibacterial compounds were further elucidated by means of ultra high-performance liquid chromatography-mass spectrometer/mass spectrometer (UHPLC-MS/MS). One compound, with a molecular mass of 883.56195 Da (M + H)+, was determined to be identical in chemical structure with that of the well-known compound fusaricidin A. The other antimicrobial compound with a molecular mass of 911.59393 Da (M + H)+ was determined to be a derivative of fusaricidin A by tandem mass spectrometry and amino acid composition analysis and was designed as bovisin. Bovisin possessed the stability against acid/alkali, heat and some proteases treatment, the same with the fusaricidin A. However, the minimal inhibition concentration (MIC) of bovisin on the tested indicator including Staphylococcus aureus, Micrococcus luteus, Listeria monocytogenes and Bacillus subtilis were 50, 50, 50, 50 μg/mL, respectively, slightly higher than those of fusaricidin A (6.25, 6.25, 6.25, 12.5 μg/mL), indicating bovisin with a weaker inhibitory activity.

A novel conotoxin, S24a, containing 26 amino acid residues was first derived from the cDNA library of Conus striatus. This conotoxin has the same conserved signal peptide as A-superfamily conotoxins and some unconserved residues in the pro-region. According to the mature peptide sequence of this conotoxin, S24a was prepared by the Escherichia coli expression system and purified by affinity chromatography, Sephadex gel filtration and reversed-phase high-performance liquid chromatography (RP-HPLC). The molecular weight of S24a was identified by MALDI-TOF-MS. Biological function experiments showed that S24a could inhibit frog sciatic nerve muscle contraction. Moreover, a high dose of S24a (100 μg/kg) had more potent and longer lasting analgesic activity compared with lidocaine and pethidine in the hotplate pain model and the formalin pain model in mice, which indicated that S24a can be further developed as a potential analgesic drug lead.

Four new prenylated flavonoids, sinoflavonoids NF–NI (1–4), were isolated from the fruits of Sinopodophyllum hexandrum by different chromatographic methods such as silica gel, Sephadex LH-20, ODS, and preparative high-performance liquid chromatography (HPLC). Their structures were elucidated on the basis of extensive spectroscopic data (UV, IR, HR-ESI-MS, 1H NMR, 13C NMR, HSQC, HMBC).

In the current work, different studies were carried out on the lipase enzyme produced by the halotolerant Oceanobacillus iheyensis strain QCS, with an expectation to be an important candidate in the industrial applications. Lipase of strain QCS was halo-alkali-thermo-detergent-solvent stable. Maximum production of lipase was obtained after 72 h of incubation, at 40oC and pH 8 and 9 in a medium containing 25 % (w/v) NaCl and 1% (v/v) olive oil as a lipid substrate. This lipase was partially purified, highest lipase activity was obtained in 80% ammonium sulfate saturation, and in fraction eight of the Sephadex G-200 gel filtration chromatography. Lipase displayed wide spectrum of activity within a broad range of conditions including salinity, temperature and pH, it was optimally active at 25% (w/v) NaCl, 40°C and pH 8 and 9, respectively. The effect of many metal ions, detergents and organic solvents on the activity of lipase was evaluated. Interestingly, lipase was able to retain the majority of its activity in the presence of Ni2+, Mg2+, Oxi and Fairy detergents, ethyl acetate, dimethyl formamide and toluene, respectively. Overall, as the lipase from O. iheyensis strain QCS has a number of interesting properties especially its stability at extreme conditions; it could be used as a potential promising candidate for detergents industry, and as a biocatalyst in low water enzymatic processes.

The bioaccumulation of keratinous wastes from poultry and dairy industries poses a danger of instability to the biosphere due to resistance to common proteolysis and as such, microbial- and enzyme-mediated biodegradation are discussed. In submerged fermentation medium, Proteus vulgaris EMB-14 utilized and efficiently degraded feather, fur and scales by secreting exogenous keratinase. The keratinase was purified 14-fold as a monomeric 49 kDa by DEAE-Sephadex A-50 anion exchange and Sephadex G-100 size-exclusion chromatography. It exhibited optimum activity at pH 9.0 and 60 °C and was alkaline thermostable (pH 7.0–11.0), retaining 87% of initial activity after 1 h pre-incubation at 60 °C. The Km and Vmax of the keratinase with keratin azure were respectively 0.283 mg/mL and 0.241 U/mL/min. Activity of P. vulgaris keratinase was stimulated by Ca2+, Mg2+, Zn2+, Na+ and maintained in the presence of some denaturing agents, except β-mercaptoethanol while Cu2+ and Pb2+ showed competitive and non-competitive inhibition with Ki 6.5 mM and 17.5 mM, respectively. This purified P. vulgaris keratinase could be surveyed for the biotechnological transformation of bioorganic keratinous wastes into valuable products such as soluble peptides, cosmetics and biodegradable thermoplastics.

Peptides from natural materials have been reported owning potential for regulating oxidative stress and hyperglycemia for human body. In this study, peptides from a favorite food, Porphyra haitanensis, were prepared by consecutive procedures including ultrasound-assisted extraction, in vitro simulated gastrointestinal digestion and fraction. Subsequently, their relevant bioactivities were investigated by several in vitro methods, such as 1,1-diphenyl-2-pycrylhydrazyl (DPPH) radical scavenging, ferric reducing antioxidant power (FRAP), and α-glucosidase enzyme inhibition assays. By single factor design experiment, the highest yield of crude protein was 3.42% extracted by alkaline solution for 20 min sonication at 35 °C. After digestion, peptides were mainly distributed in F2–F11 fractions by sephadex G-25, and the peak value was found in F5. The significant activities in three assays were displayed in F3–F7. As calculated by the polynomial curve fitting, a significantly positive correlation was found between the peptide contents and the DPPH radical scavenging rate, the FRAP value, and the α-glucosidase enzyme inhibitory rate (R2 = 0.8490, R2 = 0.7579, R2 = 0.9701, respectively). The results showed that these peptides possessed promising antioxidant activity and inhibitory activity on α-glucosidase. Therefore, peptides derived from P. haitanensis have potential anti-diabetic applications in pharmaceutical and functional food industries.

The effective utilization of okara (soybean residue) has become a considerable challenge in recent years. In this paper, the potential advantages of β-glucosidase production from okara fermented by Kluyveromyces marxianus were evaluated and the properties of the β-glucosidase were also characterized. The results showed that okara can significantly induce the production of β-glucosidase from K. marxianus. The β-glucosidase activity was up to 4.5 U/mg under optimized fermentation conditions. The optimal parameters were as follows: fermentation temperature 35 °C, cultivation time 98 h, inoculum concentration 10%, and 30 g/L of okara. After two steps of purification using ammonium sulfate precipitation and Sephadex G-75 column chromatography, the activity of β-glucosidase was 71.4 U/mg. The native enzyme was an approximately 66 kDa dimer consisting of two different subunits (22 and 44 kDa). The kinetic parameters of the K. marxianus β-glucosidase, using pNPG as substrate, were Vmax 8.34 μmol min−1 mg−1 and Km 7.42 mM. The β-glucosidase showed high thermostability and acid–alkali tolerance as well as low inhibition by DMSO (10–50%). In conclusion, this study supports the notion that okara fermentation by K. marxianus could be a useful process to produce β-glucosidase.

Objective To isolate the phenolic compounds obtained from the dried roots of Polygonum multiflorum and investigate their pharmacological activities. Methods The chemical constituents were isolated and purified by combining them with a macroporous resin (DM-8), MCI gel, and Sephadex LH-20 and by performing ODS column chromatography. Their structures were elucidated by 1D and 2D NMR analyses, as well as mass spectrometry. The isolated compounds were evaluated to determine their hepatoprotective and α-glucosidase inhibitory activities in vitro. Results Two phenolic compounds, namely, polygonimitin E (1) and polygonimitin F (2), were isolated from the dried roots of P. multiflorum. Compound 2 (10 µmol/L) only showed moderate hepatoprotective activity against N-acetyl-p-aminophenol (APAP)-induced HepG2 cell damage. Unfortunately, these two compounds exhibited no α-glucosidase inhibitory activity. Conclusion Compounds 1 and 2 were new compounds. Compound 2 could be one of the potential hepatoprotective constituents of P. multiflorum.

Purification of DPP-IV enzyme from porcine serum, is presented in this study for the first time. The high molecular weight DPP-IV from porcine serum was fractioned using Sephadex G-75 gel filtration followed by DEAE Sephadex anion exchange and Sephadex G-100 gel filtration chromatography columns with a final yield of 11.25%. The SDS-PAGE of the purified sample showed a single band of molecular mass nearing 160 kDa. Distinct single band was observed after PAS staining confirmed it to be a glycoprotein. The purified enzyme showed an optimum pH and temperature of 8 and 37 °C, respectively. The enzyme effectively cleaved fluorogenic substrate Gly-Pro-AMC with Km and Vmax of 4.578 µM and 90.84 nmoles/min, respectively. Purified DPP-IV activity was inhibited by Diprotin A with an IC50 value of 8.473 µM. Among the three plant extracts used to study DPP-IV inhibition, the aqueous hot extract of Terminalia chebula showed the highest inhibition of 87.19%, followed by the aqueous cold extract of Momordica carantia, ( 31.6%) and Azadirachta indica (34.16%) at the concentration of 25 µg.

In a recent study, anthocyanins, which have a strong free radical-scavenging activity, were examined for their potential to effectively prevent cancer. However, clinical trials are limited by the purity of the anthocyanin. Multiple methods are used to extract and purify anthocyanins. Based on previous work on Solanum nigrum, which is a widely distributed plant, in this study, DM130 macroporous resin, Sephadex LH20, and a C18 column were used to separate cis-trans anthocyanin isomers. These anthocyanins constitute the majority of total S. nigrum anthocyanins. The results showed that this "DM130-LH20-C18 system" can be used to obtain a cinnamic acid-derived cis-trans anthocyanin, petunidin-3-(p-coumaroyl)-rutinoside-5-glucoside, with a purity of 98.5%, for effective quantitation. In order to determine the antioxidant ability of the petunidin-3-(p-coumaroyl)-rutinoside-5-glucoside cis-trans isomers, three ordinary methods were adopted. The maximum antioxidant ability of the cis-trans anthocyanin was dozens of times higher than that of vitamin C.

Alkaliphilic xylanase from Neosartorya spinosa UZ-2-11 was purified using a three-step of purification scheme of ammonium sulphate precipitation followed by Sephadex G-100 gel filtration and DEAE-cellulose ion-exchange chromatography, and compared its properties with N. tatenoi KKU-CLB-3-2-4-1 of our previous report. The purified xylanase from N. spinosa UZ-2-11 exhibited maximum activity at pH 9.0 and 45 °C which was similar to endo-xylanase from N. tatenoi KKU-CLB-3-2-4-1. However, this enzyme was stable in a range of pH 6.0–11.0. It was also more stable at a high temperature of 50 °C where the activity was still up to 50% after heating for 120 min. The xylanase was purified 7.89-fold with 3.0% of yield to obtain a specific activity of 11.88 U/mg. The molecular weight of xylanase from this fungus was 27.68 kDa. The Km and Vmax values of the purified xylanase were 0.24 mg/mL and 15.85 μmol/min/mg, respectively. The xylanase activity was moderately inhibited by Hg2+ at a concentration of 10 mM, which was different to the case of N. tatenoi KKU-CLB-3-2-4-1 where Hg2+ was a strong inhibitor. In addition, the hydrolysed birchwood xylan was obtained mailnly xylobiose, xylotriose, xylotetraose and xylopentaose as end products, suggesting that it was an endo-xylanase.

The objective of this study was to examine the inhibition of the activity of enzymes associated with development of the metabolic syndrome by peptide fractions received from simulated gastrointestinal digestion and absorption of heat-treated edible insects. The inhibitory activities of insect-derived peptides were determined against key enzymes relevant to the metabolic syndrome such as the angiotensin-converting enzyme (ACE), pancreatic lipase, and α-glucosidase. After the in vitro absorption process, all hydrolysates showed high inhibitory activity; however, the most effective metabolic syndrome-inhibitory peptides were received after separation on Sephadex G10. The best results were found for peptide fractions obtained from Schistocerca gregaria. The highest enzymes inhibitory activities were obtained for peptide fractions from S. gregaria: boiled for ACE (IC50 3.95 µg mL−1), baked for lipase (IC50 9.84 µg mL−1), and raw for α-glucosiadase (IC50 1.89 µg mL−1) S. gregaria, respectively. Twelve sequences of peptides from the edible insects were identified and their chemical synthesis was carried out as well. Among the synthesized peptides, the KVEGDLK, YETGNGIK, AIGVGAIR, IIAPPER, and FDPFPK sequences of peptides exhibited the highest inhibitory activity. Generally, the heat treatment process applied to edible insects has a positive effect on the properties of the peptide fractions studied.

This study was focused on partial purification and characterization of lipase from Streptomyces acrimyciniNGP 1, isolated from marine sediment of south Indian coastal region. In purification steps, 4.53 fold purification was achieved after 85% ammonium sulphate precipitation with 0.97 percent recovery. In further purification steps, 1.33 fold purification was achieved by Sephadex G-100 chromatography with 1.61 percent of recovery. The specific activity of purified enzyme was 1525 U/mg. Zymogram of crude enzyme on native-PAGE presented bands with lipase activity of molecular weight and Isoelectric point were 50 kDa and 7.4 respectively. These features render this lipase of interest as a biocatalyst for applications such as biodiesel production and detergent formulations.

Bioactive peptides with blood pressure-lowering functions have received increasing attention. In recent years, many ACE-inhibiting peptides have been widely purified from various food-derived proteins and have received considerable interest owing to their potential role in cardiovascular diseases and in the reduction of side effects. In this study, we hydrolyzed a three-spot seahorse (Hippocampus trimaculatus Leach) protein by alcalase to obtain a hydrolysate containing angiotensin I-converting enzyme (ACE) inhibitory peptide. Then, the hydrolysate was fractionated by dialysis, Sephadex G-25 gel filtration chromatography, and reverse-phase high performance liquid chromatography. After consecutive purification, a potent ACE-inhibiting peptide composed of 8 amino acids (Pro-Ala-Gly-Pro-Arg-Gly-Pro-Ala; MW: 721.39 Da; IC(50) value: 7.90 μM) was successfully isolated from three-spot seahorse protein. For the first time, a novel ACE-inhibiting peptide (PAGPRGPA) was isolated from the seahorse. Circular dichroism (CD) analyses suggested that the secondary structure of the purified peptide was mainly composed of random coil. Therefore, the peptide from seahorse protein may be used as a favorable ingredient in nutraceuticals, medicines, and functional foods against antihypertensive and related diseases.

Although the extracellular polysaccharides have been analyzed in the previous period, the biochemical, enzymological characters and stimulation and inhibition effect on glucansucrase are not fully understood. After three steps purification, salting out, DEAE-Sepharose and Sephadex G-75, the final specific activity was 264.84 U/mg protein with 4.31-fold. The SDS-PAGE analysis of fraction gave a single band 170.35 kDa in the stained gel. The active band was analyzed with LC–MS/MS to identify glucansucrase. The highest coverage rate of dextransucrase from Leu. citreum (ACY92456.2) was 55.60%, the results were speculated that the glucansucrase secreted from Leu. citreum B-2 may be a novel glucansucrase. The purified enzyme was optimally active at 20–30 °C and pH 6.0–8.0. Metal ions K+, Na+, Ca2+, Mn2+, Mg2+, and Cr+ had an apparent stimulating effect on enzyme activity, especially in divalent ions Ca2+ and Mn2+, the residual activities were higher than 200%. In a reverse, Hg+, acetonitrile, SDS, salt, and guanidine expressed inhibition effect on enzyme residual activity. The KM and Vmax were detected to be 4.82 mM and 0.97 U/mg, respectively. All these data collectively indicate that B-2 glucansucrase is a novel one, which have good properties and may applied to new food areas.

Pesticides introduced inadvertently or deliberately into environment by anthropogenic activity have caused growing global public concern, therefore the search of approaches for elimination of such xenobiotics should be encouraged. A cypermethrin-degrading bacterial strain Bacillus licheniformis B-1 was found to efficiently degrade carbaryl in LB medium at concentrations of 50–300 mg L−1 within 48 h, during which temperature and pH played important roles as reflected by increase in pollutant depletion. A stimulatory effect of Fe3+ and Mn2+ on microbial growth was observed, whereas Cu2+ caused inhibition of degradation. Results showed that 1-naphthol was a major transformation product of carbaryl which was further metabolised. An approximately 29 kDa carbaryl-degrading enzyme was purified from B-1 with 15.93-fold purification and an overall yield of 6.02% was achieved using ammonium sulphate precipitation, DEAE-Sepharose CL-6B anion-exchange chromatography and Sephadex G-100 gel filtration. The enzyme was identified through nano reversed-phase liquid chromatography coupled with hybrid triple quadrupole time-of-flight mass spectrometry as a phosphodiesterase (PDE). This is the first report on the characterization of carbaryl-degrading by Bacillus spp. and the role of a PDE in carbaryl-detoxifying. Also, strain B-1 showed versatile in carbosulfan, isoprocarb and chlorpyrifos degradation, demonstrating as ideal candidate for environment bioremediation.

Cassava peels are waste generated from cassava processing and are mostly disposed of in Nigeria by dumping indiscriminately in landfills/waste dumps where they remain as huge mounds and constitute nuisance to the environment. This study was carried out using cassava peels as substrate for the production of α – amylase from three fungi (Aspergillus flavus, Aspergillus niger and Penicillium expansum) selected from twelve microbial isolates obtained from biodegrading cassava peels. The fungi were isolated using cassava peel agar medium and the α – amylase was produced by the solid state fermentation process. The α – amylase was assayed by measuring the decrease in staining power of starch with iodine reagent at 620nm and purified by using Sephadex G-100 and Sephadex C-50. The α – amylase was characterized by examining the effect of temperature, stability at 70oC, pH, substrate concentration, metal ions and EDTA. The results show that the purification fold and specific activity were 95.727, 1.053 Units/mg protein; 81.830, 0.982 Units/mg protein and 85.784, 0.686 Units/mg protein for A. flavus, A. niger and P. expansum respectively. The optimum temperature and pH were 45oC and 4.5 respectively. It was observed that the α – amylase still retained some activity after heating at 70oC for 35 min. The α – amylase activity increased with increase in substrate concentration and metal ion concentration (Na+, K+, Mg2+ and Ca2+) but decreased with increase in heavy metal ion concentration (Hg2+ and Pb2+) and EDTA. Key words: Cassava peels, α – amylase, fungi, starch and Sephadex C – 50.

A Box-Behnken design was employed to optimize ultrasound-assisted enzymatic extraction of polysaccharides from perilla leaves. Four independent variables, namely, liquid-to-solid ratio, enzymatic time, enzymatic temperature, and ultrasonic power were investigated. Under the optimal liquid-to-solid ratio of 41:1, enzymatic time of 40 min, enzymatic temperature of 49 °C, and ultrasonic power of 204 W, the experimental polysaccharide yield was 3.84% (n = 3), which was close to the value predicted by response surface methodology (3.9%). Four polysaccharides (PLP1, PLP2, PLP3, and PLP4) were obtained with DEAE-cellulose-52 and Sephadex G-100 chromatography, and their antioxidant activities were investigated with various antioxidant assays in vitro. All the four polysaccharides possessed good antioxidant properties in a concentration-dependent manner, and PLP3 presented the highest antioxidant activity. Thus, the novel polysaccharides extracted from perilla leaves could be promising bioactive macromolecules that can be applied as potential antioxidants in medical and food industries.

Highlights • Ethyl acetate extraction resulted in the highest bikaverin yield. • Kinetic study revealed a saturation of bikaverin extraction after 256 min. • Three sequential ethyl acetate extractions was the most economical to recover bikaverin. • Open column chromatography or nanofiltration were not suitable to purify bikaverin. • Bikaverin was successfully purified on semi-preparative HPLC., Microbial pigments have a distinguished potential for applications in food and pharmaceutical industries, stimulating the research in this field. The present study evaluated the ideal conditions for extracting bikaverin (red pigment) from the biomass of Fusarium oxysporum CCT7620. Among the solvents tested, ethyl acetate extraction resulted in the highest bikaverin concentration and the kinetic study revealed a saturation in bikaverin concentration from 256 min on. Based on a preliminary economic study, three sequential extractions with ethyl acetate was considered the ideal protocol to recover bikaverin. After extraction, chromatographic methods were tested to purify bikaverin. The use of silica gel or Sephadex (open column) could not successfully purify bikaverin, but the semi-preparative HPLC resulted in a bikaverin-enriched fraction with a purity degree equivalent to the commercial analytical standard. This work provides relevant information regarding the extraction and purification of bikaverin, which may be useful for other downstraming processes.

Alcalase, neutral protease, and pepsin were used to hydrolyze the skin of Takifugu flavidus. The T. flavidus hydrolysates (TFHs) with the maximum degree of hydrolysis (DH) and angiotensin-I-converting enzyme (ACE)-inhibitory activity were selected and then ultra-filtered to obtain fractions with components of different molecular weights (MWs) (50 kDa). The components with MWs < 1 kDa showed the strongest ACE-inhibitory activity with a half-maximal inhibitory concentration (IC50) of 0.58 mg/mL. Purification and identification using semi-preparative liquid chromatography, Sephadex G-15 gel chromatography, RP-HPLC, and LC–MS/MS yielded one new potential ACE-inhibitory peptide, PPLLFAAL (non-competitive suppression mode; IC50 of 28 μmmol·L−1). Molecular docking and molecular dynamics simulations indicated that the peptides should bind well to ACE and interact with amino acid residues and the zinc ion at the ACE active site. Furthermore, a short-term assay of antihypertensive activity in spontaneously hypertensive rats (SHRs) revealed that PPLLFAAL could significantly decrease the systolic blood pressure (SBP) and diastolic blood pressure (DBP) of SHRs after intravenous administration. These results suggested that PPLLFAAL may have potential applications in functional foods or pharmaceuticals as an antihypertensive agent.

β-amylase-rich preparation produced by the soil fungal strain Penicillium citrinum AS-9 was completely purified within the succeeding steps; ultrafiltration, acetone fractionation and gel filtration on Sephadex G-150 column. Acetone fractionation recovered the highest β-amylase activity and the 85% acetone fraction exhibited 3.5-fold activity that of the crude enzyme. The column affected 8.6-fold purification for β-amylase, which produced maltose as the major product of starch hydrolysis. The pure enzyme showed a Km value of 17.5 mM and Vmax of 17.5 Umg-1 protein, applying Woolf plot and exhibited its maximum velocity at pH 7.1 and 50°C. In absence of substrate, thermal treatments of the enzyme solution at pH 5.2 and 5.5 had the most adverse effects on the enzyme activity. CaCl2 (1mM) activated the enzyme, while each of Ca2+, Fe3+, cysteine, cystine, I2 and p-chloromercuribenzoate (PCMB) had different inhibitory effects. Maltose as the enzyme product at final concentration of 80 mM strongly inhibited the enzyme.

Soybean cyst nematode (SCN) (Heterodera glycines Ichinohe) is responsible for causing a major soybean disease globally. The fungal strain Penicillium janthinellum Snef1650 was evaluated against H. glycines. However, the effective determinants of the P. janthinellum strain are unknown. By performing pot experiments, a functioning compound was isolated from P. janthinellum Snef1650 through organic solvent extraction, semi-preparative HPLC, Sephadex LH-20 column chromatography, and silica gel column chromatography, and the isolated compound was identified to be scopoletin through 1H NMR, 13C NMR, and HPLC–MS. The pot experiments indicated that the treatment of soybean seeds with scopoletin drastically reduced the SCN population. The field experiments performed in 2017 and 2018 revealed that scopoletin decreased over 43.7% juveniles in the roots and over 61.55% cysts in the soil. Scopoletin treatment also promoted soybean growth and improved its yield, with an increase in plot yield by &gt, 5.33%. Scopoletin obtained from P. janthinellum Snef1650 could be used as an anti-H. glycines biocontrol agent.

Nepeta bracteata Benth. is used clinically to treat tracheal inflammation, coughs, asthma, colds, fevers, adverse urination, and other symptoms, along with functions in clearing heat and removing dampness. However, there have been few studies characterizing the material basis of its efficacy. Therefore, the aim of this study was to screen for compounds with anti-inflammatory activities in N. bracteata Benth. Using silica gel, ODS C18, and Sephadex LH-20 column chromatography, as well as semipreparative HPLC, 10 compounds were separated from&nbsp, N. bracteata Benth. extract, including four new diterpenoids (1–4), one amide alkaloid (5), and five known diterpenoids (6–10). The structures of all the isolates were elucidated by HR-ESI-MS, NMR, and CD analyses. Using lipopolysaccharide (LPS)-stimulated RAW&nbsp, 264.7 cells, we investigated the anti-inflammatory&nbsp, activities of compounds 1–10. It is worth noting that all were able to inhibit nitric oxide (NO) production with IC50 values &lt, 50 μM and little effect on RAW 264.7 macrophage viability. Compounds 2 and 4 displayed remarkable inhibition with IC50 values of 19.2 and 18.8 μM, respectively. Meanwhile, screening on HCT-8 cells demonstrated that compounds 2 and 4 also had moderate cytotoxic activities with IC50 values of 36.3 and 41.4 μM, respectively, which is related to their anti-inflammatory effects.

Synsepalum msolo commonly known as Bang Bali in Bali-Nguemba, Cameroon is used in traditional medicine against various diseases. The leaves and stem bark extracts were subjected to silica gel and Sephadex LH20 column chromatography to yield pure compounds. The structures of the compounds were determined by detail analysis of NMR and Mass spectroscopic data and by comparison with data reported in the literature. Amongst the isolates, were two new sphingolipids: synsepaloside B (1), synsepaloside C (2), and five known compounds: (+)-catechin (3), (−)-epicatechin (4), myricitrin (5), triacontanol (6), and aurantiamide acetate (7). Compounds 1–5 were screened for their antibacterial and anti-yeast activities on several microorganisms. All the tested compounds exhibited weak antibacterial (MIC ≥ 200 μg/mL) and anti-yeast (MIC > 200 μg/mL) activities as compared to standard: ciprofloxacin 0.468 0.234 μg/mL and fluconazole MIC = 0.05 μg/mL, respectively.