Microbiological Transformation of L-Tyrosine to L-Dopa from Methanol Pretreated Biomass of a Novel Coriolus versicolor under Submerged Culture

The present study is concerned with the microbiological transformation of L-tyrosine to L-dopa by a newly isolated turkey tail mushroom Coriolus versicolor DOB-4. As tyrosinase (catechol oxidase, EC 1.10.3.1) is an extracellular enzyme, therefore biomass was used as an enzyme source in the reaction mixture. Biomass particles were pretreated with methanol and oven dried at 105 °C for 2 h. The optimal L-dopa production was achieved when 1.5 mg/ml L-tyrosine was used as the basal substrate. Thin layer chromatography and high-performance liquid chromatography analysis depicted that citric acid supports higher substrate conversion and product formation rates. A noticeable enhancement was observed when process parameters viz. L-tyrosine concentration (1.5 mg/ml), citric acid (1.5 mg/ml), time of incubation (50 min), and reaction temperature (60 °C) were optimized using Plackett–Burman design. The maximum production of L-dopa was found to be 0.872 mg/ml with L-tyrosine consumption of 1.002 mg/ml. The model terms were found highly significant (HS, p?≤?0.05), suggesting the potential commercial utility of the culture (df?=?3, LSD?=?0.342).     

Electrospinning of nylon-6,6 solutions into nanofibers: Rheology and morphology relationships

The relationship between the rheological properties of nylon-6,6 solutions and the morphology of their electrospun nanofibers was established. The viscosity of nylon-6,6 in formic acid(90%) was measured in the concentration range of 5 wt%-25 wt% using a programmable viscometer. Electrospinning of nylon-6,6 solutions was carried out under controlled parameters. The chemical structure, morphology and thermal properties of the obtained nanofibers were investigated using Fourier transform infrared spectroscopy(FTIR), scanning electron microscopy(SEM) and differential scanning calorimetry(DSC), respectively. Entanglement concentration(ce) was found to be 15 wt% and a power law relationship between specific viscosity and solution concentration was observed with exponents of 2.0 and 3.3 for semi-dilute unentangled(c ce) and semi-dilute entangled(c ce) regimes, respectively. The diameter and uniformity of the nanofibers were found to be dependent on the viscosity. Moreover, the average diameter of electrospun nanofibers was found to be dependent on zero shear rate viscosity and normalized concentration(c/ce) in a power law relationship with exponents of 0.298 and 0.816, respectively. For nylon-6,6 solutions, the entanglement concentration(ce = 15 wt%) provides the threshold viscosity required for the formation of a stable polymeric jet during electrospinning and producing uniform beadless fibers. For concentrations less than ce, beaded fibers with some irregularities are formed. DSC analysis showed an increase in crystallinity of all electrospun samples compared to original polymer. Furthermore, Based on FTIR spectroscopy, α phase is dominant in electrospun nanofibers and minor amount of β and γ phases is also available.     

Improvements of Tolerance to Stress Conditions by Genetic Engineering in Saccharomyces Cerevisiae during Ethanol Production

Saccharomyces cerevisiae, industrial yeast isolate, has been of great interest in recent years for fuel ethanol production. The ethanol yield and productivity depend on many inhibitory factors during the fermentation process such as temperature, ethanol, compounds released as the result of pretreatment procedures, and osmotic stress. An ideal strain should be able to grow under different stress conditions occurred at different fermentation steps. Development of tolerant yeast strains can be achieved by reprogramming pathways supporting the ethanol metabolism by regulating the energy balance and detoxicification processes. Complex gene interactions should be solved for an in-depth comprehension of the yeast stress tolerance mechanism. Genetic engineering as a powerful biotechnological tool is required to design new strategies for increasing the ethanol fermentation performance. Upregulation of stress tolerance genes by recombinant DNA technology can be a useful approach to overcome inhibitory situations. This review presents the application of several genetic engineering strategies to increase ethanol yield under different stress conditions including inhibitor tolerance, ethanol tolerance, thermotolerance, and osmotolerance.     

Insulin Resistance Associated Genes and miRNAs

Type 2 diabetes mellitus is the result of resistance to insulin function along with inadequate insulin secretion, leading to a number of dysfunctions characterized by hyperglycemia, and it is associated with microvascular, macrovascular, and neuropathic complications. There is compelling evidence that the decline in both insulin sensitivity and insulin secretion has a genetic component. In addition, increasing evidence suggests that microRNAs (miRNAs) as key regulators of gene expression play significant roles in insulin production, secretion, and function that regulate the function of insulin-target tissues. The current review demonstrates the candidate genes and the related miRNAs involved in molecular pathogenesis of insulin resistance in type 2 diabetes mellitus. In doing so, it provides an opportunity for more focused investigations that may identify the genes and miRNAs with a role in the pathogenesis of type 2 diabetes mellitus and its treatment.     

Chitosan-Modified Carbon Nanotubes-Based Platform for Low-Density Lipoprotein Detection

We have fabricated an immunosensor based on carbon nanotubes and chitosan (CNT-CH) composite for detection of low density lipoprotein (LDL) molecules via electrochemical impedance technique. The CNT-CH composite deposited on indium tin oxide (ITO)-coated glass electrode has been used to covalently interact with anti-apolipoprotein B (antibody: AAB) via a co-entrapment method. The biofunctionalization of AAB on carboxylated CNT-CH surface has been confirmed by Fourier transform infrared spectroscopic and electron microscopic studies. The covalent functionalization of antibody on transducer surface reveals higher stability and reproducibility of the fabricated immunosensor. Electrochemical properties of the AAB/CNT-CH/ITO electrode have been investigated using cyclic voltammetric and impedimetric techniques. The impedimetric response of the AAB/CNT-CH/ITO immunoelectrode shows a high sensitivity of 0.953?Ω/(mg/dL)/cm² in a detection range of 0–120 mg/dL and low detection limit of 12.5 mg/dL with a regression coefficient of 0.996. The observed low value of association constant (0.34 M^–1s^–1) indicates high affinity of AAB/CNT-CH/ITO immunoelectrode towards LDL molecules. This fabricated immunosensor allows quantitative estimation of LDL concentration with distinguishable variation in the impedance signal.     

Biogas Production from N-Methylmorpholine-N-oxide (NMMO) Pretreated Forest Residues

Lignocellulosic biomass represents a great potential for biogas production. However, a suitable pretreatment is needed to improve their digestibility. This study investigates the effects of an organic solvent, N-Methylmorpholine-N-oxide (NMMO) at temperatures of 120 and 90 °C, NMMO concentrations of 75 and 85 % and treatment times of 3 and 15 h on the methane yield. The long-term effects of the treatment were determined by a semicontinuous experiment. The best results were obtained using 75 % NMMO at 120 °C for 15 h, resulting in 141 % increase in the methane production. These conditions led to a decrease by 9 % and an increase by 8 % in the lignin and in the carbohydrate content, respectively. During the continuous digestion experiments, a specific biogas production rate of 92 NmL/gVS/day was achieved while the corresponding rate from the untreated sample was 53 NmL/gVS/day. The operation conditions were set at 4.4 gVS/L/day organic loading rate (OLR) and hydraulic retention time (HRT) of 20 days in both cases. NMMO pretreatment has substantially improved the digestibility of forest residues. The present study shows the possibilities of this pretreatment method; however, an economic and technical assessment of its industrial use needs to be performed in the future.     

Trypsin Inhibitor from Edible Mushroom Pleurotus floridanus Active against Proteases of Microbial Origin

Protease inhibitors can be versatile tools mainly in the fields of medicine, agriculture and food preservative applications. Fungi have been recognized as sources of protease inhibitors, although there are only few such reports on mushrooms. This work reports the purification and characterization of a trypsin inhibitor from the fruiting body of edible mushroom Pleurotus floridanus (PfTI) and its effect on the activity of microbial proteases. The protease inhibitor was purified up to 35-fold by DEAE-Sepharose ion exchange column, trypsin-Sepharose column and Sephadex G100 column. The isoelectric point of the inhibitor was 4.4, and its molecular mass was calculated as 37 kDa by SDS-PAGE and 38.3 kDa by MALDI-TOF. Inhibitory activity confirmation was by dot-blot analysis and zymographic activity staining. The specificity of the inhibitor toward trypsin was with Ki of 1.043?×?10^?10 M. The inhibitor was thermostable up to 90 °C with maximal stability at 30 °C, active over a pH range of 4–10 against proteases from Aspergillus oryzae, Bacillus licheniformis, Bacillus sp. and Bacillus amyloliquefaciens. Results indicate the possibility of utilization of protease inhibitor from P. floridanus against serine proteases.     

Purification and Characterization of Pepsin-Solubilized Collagen from Skin of Sea Cucumber Holothuria parva

Pepsin-solubilized collagen (PSC) was extracted from the skin of sea cucumber Holothuria parva and was fractionally characterized. The PSC from H. parva skin consisted of three α₁ chains (α₁)₃, in contrast to calf skin collagen type I with two α₁ and one α₂ chains (α₁)₂α₂ with approximately 130 kDa each. The maximum transition (T_m) and denaturation temperature (T_d) of PSC were determined to be 46.94 and 32.5 °C, respectively. The amino acid composition analysis revealed that glycine, proline, alanine, and hydroxyproline were the abundant amino acids available in extracted PSC. The results showed that the isolated collagen from H. parva has some similar characteristics to previously reported collagens used in food and pharmaceutical industries.     

A new type of monodisperse porous, hydrophilic microspheres with reactive chloroalkyl functionality: synthesis and derivatization properties

A seeded polymerization method based on a new functional monomer, 3-chloro-2-hydroxypropyl methacrylate (HPMA-Cl), was proposed for the synthesis of a new type of monodisperse porous, hydrophilic microspheres with reactive character. By applying the method, poly(3-chloro-2-hydroxypropyl methacrylate-co-ethylene dimethacrylate) (poly(HPMA-Cl-co-EDMA)) microspheres in the range of 4–7 μm, with specific surface areas between 2 and 146 m²/g, were obtained. The microspheres are hydrophilic in nature due to the hydroxyl groups and are easily derivatizable due to the reactive chloropropyl moiety. Ligands in the form of small molecules carrying hydrophobic alkyl or hydrophilic ion exchanger groups were covalently attached onto the microspheres via simple and one-pot reactions via their chloropropyl functionality. Using the same functionality, click chemistry and surface-initiated atom transfer radical polymerization were also applied for the generation of triazole ring and zwitterionic molecular brushes on the microspheres, respectively. Poly(HPMA-Cl-co-EDMA) microspheres seem to be a promising hydrophilic reactive material particularly for the synthesis of ion exchanger resins and chromatographic stationary phases.