Influence of Relative Molecular Mass of SBS on Low-Temperature Performance of Modified Asphalt

Styrene Butadiene Styrene (SBS) is widely used in pavements because it significantly improves the performance of the modified asphalt. The dosage of SBS has an important effect on the low-temperature performance of the modified asphalt. The difference in relative molecular weight influences the magnitude of the change in low-temperature performance. In this paper, low temperature bending beams rheology test of five different relative molecular weight SBS modified asphalt is conducted at -12°C, -18°C and -24°C to evaluate the low-temperature properties of modified asphalt. Then, the experimental results are analysed. The results show that there is a certain correlation between the relative molecular mass of SBS and the low-temperature performance of modified asphalt. The creep stiffness and creep rate of the modified asphalt are influenced by the relative molecular mass of SBS and temperature. As the relative molecular mass increases, the low-temperature performance of the modified asphalt first rises and then decreases.

Bacillus velezensis Identification and Recombinant Expression, Purification and Characterization of Its Alpha-Amylase

Amylases account for about 30% of the global market of industrial enzymes, and the current amylases cannot fully meet industrial needs. This study aimed to identify a high α-amylase producing bacterium WangLB, to clone its α-amylase coding gene, and to characterize the α-amylase. Results showed that WangLB belonged to Bacillus velezensis whose α-amylase gene was 1980 bp coding 659 amino acids designated as BvAmylase. BvAmylase was a hydrophilic stable protein with a signal peptide and a theoretical pI of 5.49. The relative molecular weight of BvAmylase was 72.35 kDa, and was verified by SDS-PAGE. Its modeled structure displayed that it was a monomer composed of three domains. Its optimum temperature and pH were 70 °C and pH 6.0, respectively. It also showed high activity in a wide range of temperatures (40–75 °C) and a relatively narrow pH (5.0–7.0). It was a Ca2+-independent enzyme, whose α-amylase activity was increased by Co2+, Tween 20, and Triton X-100, and severely decreased by SDS. The Km and the Vmax of BvAmylase were 3.43 ± 0.53 and 434.19 ± 28.57 U/mg. In conclusion, the α-amylase producing bacterium WangLB was identified, and one of its α-amylases was characterized, which will be a candidate enzyme for industrial applications.

Molecular characterization and enzyme inhibition studies of NADP+- farnesol dehydrogenase from diamondback moth, Plutella xylostella (Lepidoptera: Plutellidae)

Juvenile hormone III (JH III) plays an important role in insect reproduction, development, and behavior. The second branch of JH III production includes oxidation of farnesol to farnesal by farnesol dehydrogenase. This study reported the identification and characterization of Plutella xylostella farnesol dehydrogenase (PxFoLDH). Our results showed that PxFoLDH belongs to the short-chain dehydrogenase/reductase superfamily, consisting of a single domain with a structurally conserved Rossman fold, an NAD(P) (H)—binding region and a structurally diverse C- terminal region. The purified enzyme displayed maximum activity at 55 °C with pH 9.5 and was stable in the temperature below 70 °C. PxFoLDH was determined to be a monomer with a relative molecular weight of 27 kDa and highly specific for trans, trans-farnesol and NADP+. Among analog inhibitors tested, farnesyl acetate was the most effective inhibitor with the lowest Ki value of 0.02 µM. Our findings showed this purified enzyme may represent as NADP+-farnesol dehydrogenase.

EPS364, a Novel Deep-Sea Bacterial Exopolysaccharide, Inhibits Liver Cancer Cell Growth and Adhesion

The prognosis of liver cancer was inferior among tumors. New medicine treatments are urgently needed. In this study, a novel exopolysaccharide EPS364 was purified from Vibrio alginolyticus 364, which was isolated from a deep-sea cold seep of the South China Sea. Further research showed that EPS364 consisted of mannose, glucosamine, gluconic acid, galactosamine and arabinose with a molar ratio of 5:9:3.4:0.5:0.8. The relative molecular weight of EPS364 was 14.8 kDa. Our results further revealed that EPS364 was a β-linked and phosphorylated polysaccharide. Notably, EPS364 exhibited a significant antitumor activity, with inducing apoptosis, dissipation of the mitochondrial membrane potential (MMP) and generation of reactive oxygen species (ROS) in Huh7.5 liver cancer cells. Proteomic and quantitative real-time PCR analyses indicated that EPS364 inhibited cancer cell growth and adhesion via targeting the FGF19-FGFR4 signaling pathway. These findings suggest that EPS364 is a promising antitumor agent for pharmacotherapy.

Preparation, Preliminary Characterization and Antiangiogenic Activities of Polysaccharides from Pomegranate Peels

In the present study, the crude polysaccharides from pomegranate peels (CPP) were prepared by compound enzyme-extraction technology, and the response surface methodology was used to optimize the extraction parameters. Box–Behnken design (BBD) was applied to estimate the effects of extraction temperature, extraction pH, and dosage of compound enzyme on the yield of CPP. A mathematical model with high fitness was obtained. Extraction temperature, extraction pH, and dosage of compound enzyme exhibited independent and interactive effects on CPP yield. CPP were purified by macroporous resin HP-20 and Sephadex G-100 column chromatography to afford purified fraction of CPP-2. The relative molecular weight of CPP-2 was 93.5 kDa. In CPP-2, and FT-IR spectra showed that the main components among CPP-2 may be pectic polysaccharides.The effect of CPP-2 on angiogenesis was measured in vivo by using the chick embryo chorioallantoic membrane (CAM) assay. The results demonstrated that CPP-2 suppressed angiogenesis in chicken embryos.

Anti-osteoporosis effect and purification of peptides with high calcium-binding capacity from walnut protein hydrolysate

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. Results showed that...

Identification of a Novel Thermostable Alkaline Protease from Bacillus megaterium-TK1 for the Detergent and Leather Industry

An increased need by the green industry for enzymes that can be exploited for eco-friendly industrial applications led us to isolate and identify a unique protease obtained from a proteolytic Bacillus megaterium-TK1 strain from a seawater source. The extracellular thermostable serine protease was processed by multiple chromatography steps. The isolated protease displayed a relative molecular weight (MW) of 33 kDa (confirmed by zymography), optimal enzyme performance at pH 8.0, and maximum enzyme performance at 70 °C with 100% substrate specificity towards casein. The proteolytic action was blocked by phenylmethylsulfonyl fluoride (PMSF), a serine hydrolase inactivator. Protease performance was augmented by several bivalent metal cations. The protease tolerance was studied under stringent conditions with different industrial dispersants and found to be stable with Surf Excel, Tide, or Rin detergents. Moreover, this protease could clean blood-stained fabrics and showed dehairing activity for cow skin with significantly reduced pollution loads. Our results suggest that this serine protease is a promising additive for various eco-friendly usages in both the detergent and leather industries.

UHPLC Q-Exactive MS-Based Serum Metabolomics to Explore the Effect Mechanisms of Immunological Activity of Astragalus Polysaccharides With Different Molecular Weights

Astragalus polysaccharides (APS) have a wide range of biological activities. Most researchers discuss total APS as the main research object. However, because the relative molecular weight of APS has a wide distribution, in-depth studies on the mechanisms of the biological activity of notable molecules are limited. For example, the relationship between the immunomodulatory effect of APS and its relative molecular weight has not been clearly defined. Therefore, in this paper, we separated and obtained APS of different molecular weights by ultrafiltration technology and then constructed a mouse cyclophosphamide-induced immunosuppression model to investigate the immune activity of APS of different molecular weights. The immune enhancement mechanism of APS was explored by examining changes in routine blood indicators, body weight, immune organs, and differential metabolites in mouse serum. Results showed that APS-I (molecular weight, >2,000 kDa), APS-II (molecular weight, 1.02 × 104 Da) and APS-III (molecular weight, 286 Da) could increase the number of immune cells in mouse serum and improve immune organ damage to varying degrees. Among the samples obtained, APS-II showed the best effects. Compared with those in the blank group, 29 metabolites determined by UHPLC Q-Exactive MS in the serum of the model group changed remarkably, and APS-I, APS-II, and APS-III respectively restored 13, 25, and 19 of these metabolites to normal levels. Metabolomics analysis revealed that APS-II is mainly responsible for the immunomodulatory activity of APS. Metabolomics analysis revealed that the mechanisms of this specific molecule may involve the regulation of phenylalanine metabolism, cysteine and methionine metabolism, tricarboxylic acid cycle (TCA cycle) and arginine and proline metabolism.

Laboratory Study of Flocculation and Pressure Filtration Dewatering of Waste Slurry

In the process of large-scale urban construction, large amounts of waste slurry are produced. The slurry has a high water content and is difficult to precipitate naturally, resulting in low treatment efficiency. To improve the treatment efficiency of slurry, a variety of inorganic and organic polymer flocculants were used to carry out flocculation settlement tests on the slurry. The changes in the slurry properties and the filtration dewatering effect after flocculation were tested. The results show that the addition of flocculant makes the slurry particles form aggregates, which leads to rapid precipitation of the slurry. The use of an inorganic flocculant significantly reduced the zeta potential of the slurry. Organic polymer flocculant, however, had little effect on the slurry potential, but did cause the slurry to produce larger size aggregates, resulting in a better flocculation effect than inorganic flocculant. Inorganic flocculants and organic flocculants can improve the pressure filtration dewatering performance of slurry. CPAM12 (cationic polyacrylamide, with a relative molecular weight of 12 million Daltons) had the best overall effect. The formation of aggregates after flocculation and the change in the nonuniformity coefficient (Cu) were the main cause of improvement of pressure filtration dewatering performance of the slurry. When Cu decreases from 11.85 to 8.75, the time required for pressure filtration stabilization is shortened by 70%. The nonuniformity coefficient of flocculated slurry can be used to evaluate flocculants, determine the optimal dosage, and predict the dewatering effect.

The expression and purification of LpxA of Chlamydia trachomatis and preparation of its polyclonal antibody

The purpose of this study is to purify the LpxA protein of Chlamydia trachomatis (Ct) and prepare the polyclonal antibody against LpxA protein, so as to lay a foundation for studying the function of LpxA protein. The LpxA gene was amplified by PCR. The expression plasmid pET28a-LpxA was constructed by using pET28a as the vector. The fusion protein containing 6 histidine tag was induced by IPTG and purified by Ni2+ chromatography gel. The purified His-LpxA protein was used as an immunogen to immunize New Zealand rabbits subcutaneously through the back to prepare polyclonal antibody. Immunoblotting was used to detect the reaction between the antibody and His-LpxA. The determination of polyclonal antibody titer was detected by ELISA. The relative molecular weight of His-LpxA was 32.8 kDa, and it could be expressed in Escherichia coli. The purity of the purified protein was about 95%. After immunizing New Zealand rabbits, the antiserum was able to recognize the recombinant His-LpxA protein with a titer greater than 1:10240. In this study, LpxA protein was successfully purified and antiserum was prepared, which provided an experimental basis for studying the function of LpxA protein.