Characterization of recombinant L-ribose isomerase acquired from Cryobacterium sp. N21 with potential application in L-ribulose production

l-Ribose isomerase (lRI) is an enzyme that can catalyze the reversible isomerization between l-ribose and l-ribulose. It can also perform the conversion between many aldoses into their corresponding ketoses. l-RI was produced from Cryobacterium sp. N21 (CrL-RIse), and l-ribose was utilized as a substrate. The recombinant l-RI gene was cloned and overexpressed from Cryobacterium sp. N21. The purification of CrL-RIse was performed by metal-affinity chromatography. The enzyme displayed a corresponding band with an approximate size of 35 kDa on the SDS-PAGE analysis. The protein for this gene contains 266 amino acids with an expected molecular weight (M_w) of 29.6 kDa. The measured M_w of CrL-RIse calculated by HPLC was 125 kDa. CrL-RIse was extremely active in glycine buffer at 35 °C, pH 9.0, showing a specific activity of 54.96 U mg⁻¹. CrL-RIse displayed no major increase in activity with metal ions, excluding Mn²⁺. The estimated Km, K_cat, K_cat/Km and V_max values of CrL-RIse were 37.8 mM, 10,416 min⁻¹, 275.43 min⁻¹ mM⁻¹, and 250 U mg⁻¹, respectively. The rate of l-ribulose production was 31 % (6.24, 12.11, and 20.89 g L⁻¹) at equilibrium by utilizing 20, 40, and 70 g L⁻¹ of the substrate, respectively. The results indicated that CrL-RIse has the capability to manufacture l-ribulose from l-ribose.     

SYNTHESIS AND CHARACTERIZATION OF ADENOSINE ADDUCTS OF ARYLAMINES

ABSTRACT

The synthesis of adducts of arylamines with adenosine are reported.     

Biosorption of lead by cotton shells powder: Characterization and equilibrium modeling study

Lead (Pb) is a toxic heavy metal causing serious health risks to humans and animals. In the present study, cotton (Gossypium hirsutum L.) shells powder was used as adsorbent for the treatment of synthetic Pb-contaminated water. The batch scale biosorption capacity of cotton shells powder was evaluated to study the effects of Pb concentrations, adsorbent doses and contact time at constant pH (6) and temperature (25?°C). Results revealed that sorption of Pb increased (q?=?0.09–9.60?mg/g) with increasing Pb concentration (1–15?mg/L) and contact time (15–90?min) while decreasing adsorbent dose (1–0.1?g/100?mL). The maximum Pb removal (90%) was achieved at Pb concentration (1?mg/L), contact time (90?min) and adsorbent dose (1?g/100?mL). Freundlich isotherm model proved best fit for Pb sorption (R²?=?0.99). The cotton shells powder has microporous structure confirmed by SEM, and has BET surface area (45 m²/g) and pore size (2.3 µm). These surface moieties along with various functional groups (C-H, C-O, C=O, O-H, S=O) confirmed by FTIR analysis might involve in Pb removal by complexation and ion exchange mechanisms. The cotton shells powder biomass could be considered as promising adsorbent for the removal of Pb from contaminated water.     

Ag/TiO2纳米材料对烟曲霉的抑制作用及机制

目的合成Ag/TiO_2纳米材料,对其进行表征测定,并探讨其对烟曲霉的抑制作用及具体机制。方法采用光催化还原法制备Ag/TiO_2纳米材料,紫外可见分析和扫描电镜对其进行表征测定;微量液基稀释法检测对烟曲霉的最低抑菌浓度(MIC),以及生物量的抑制作用;ELISA试剂盒检测对真菌谷胱甘肽还原酶、总谷胱甘肽、线粒体膜电位的影响,荧光显微镜检测活性氧的产生。结果成功制备Ag/TiO_2纳米材料,分布均匀;对烟曲霉的MIC值为0.5μg/mL,能完全抑制烟曲霉生物量,与单独纳米银相比,具有更好的抗菌活性。机制研究发现其主要通过降低烟曲霉体内谷胱甘肽及其还原酶的含量,诱导过量活性氧的产生,最终导致线粒体膜电位降低,使真菌细胞发生凋亡。结论 Ag/TiO2纳米材料可有效阻断烟曲霉等真菌在空气中的传播,具有广泛的应用前景。     

Reproductive biology and morphology of Apis mellifera jemenitica (Apidae) queens and drones

The current study aimed to investigate the important reproductive biology and morphology of A.m. jemenitica queens and drones through measuring the weight of virgin and mated queens, size and weight of spermathecae, weight of ovaries, number of ovarioles, quantity and viability of semen in queen and drones. Accordingly, the average weights of 0.139 ± 0.01 g and 0.143 ± 0.013 g recorded for virgin and mated queens respectively. The sizes of spermathecae were 1.248 ± 0.103 mm and 1.25 ± 0.022 mm for virgin and mated queens respectively. The mean weight of ovaries was 0.013 ± 0.003 g and the numbers of ovarioles varied from 124 to 163 with the mean of 142.9 ± 9.47 and with no significant difference between virgin and mated queens. The average number of stored sperm per spermathecae of mated queen was estimated to be 4.202 ± 0.613 million with the viability of 80.39%. The average number of sperm per drone recorded was 8,763,950 ± 1,633,203.15 with viability of 79.54 ± 6.70%. In general, the current study revealed that the values recorded for reproductive biology and morphological characters of A. m. jemenitica queens and drones were relatively lower than values recorded for other Apis mellifera races. This mainly could be associated with the body size of the race which is known to be the smallest race among A. mellifera races. Moreover, the harsh environmental conditions of the regions, high temperature, low humidity and limited resources may have contributed for the smaller biological and morphological values. The information will serve as a base in future selection and breeding of program of the race.     

A bone preservation protocol that enables evaluation of osseointegration of implants with micro- and nanotextured surfaces

Development of surface treatments has enabled secure attachment of dental implants in less than 1 month. Consequently, it is necessary to characterize accurately the osseointegration of the implant surface in the region of the bone-implant contact (BIC). We developed a method for sample preparation that preserves both bone and BIC to permit analysis of the contact interface. We prepared eight nanotextured implants and implanted them in rabbit tibias. After healing for 30 days, outcomes were analyzed using both our bone preservation protocol and routine decalcification followed by preparation of histological sections stained by hematoxylin and eosin (H & E). Pull-out tests for implant osseointegration were performed after healing. Non-implanted samples of rabbit mandible were used as a control for assessing organic and mineralized bone characteristics and bone structure. Our bone preservation protocol enabled evaluation of many of the same bone characteristics as histological sections stained with H & E. Our protocol enables analysis of implant samples, implant surfaces and osseointegration without risk of BIC damage.     

A rapid approach for characterization of thiol-conjugated antibody–drug conjugates and calculation of drug–antibody ratio by liquid chromatography mass spectrometry

We present the demonstration of a rapid “middle-up” liquid chromatography mass spectrometry (LC–MS)-based workflow for use in the characterization of thiol-conjugated maleimidocaproyl-monomethyl auristatin F (mcMMAF) and valine-citrulline-monomethyl auristatin E (vcMMAE) antibody–drug conjugates. Deconvoluted spectra were generated following a combination of deglycosylation, IdeS (immunoglobulin-degrading enzyme from Streptococcus pyogenes) digestion, and reduction steps that provide a visual representation of the product for rapid lot-to-lot comparison—a means to quickly assess the integrity of the antibody structure and the applied conjugation chemistry by mass. The relative abundance of the detected ions also offer information regarding differences in drug conjugation levels between samples, and the average drug–antibody ratio can be calculated. The approach requires little material (<100 μg) and, thus, is amenable to small-scale process development testing or as an early component of a complete characterization project facilitating informed decision making regarding which aspects of a molecule might need to be examined in more detail by orthogonal methodologies.