Optimisation of growth medium for the production of cyclodextrin glucanotransferase from Bacillus stearothermophilus HR1 using response surface methodology

Cyclodextrin glucanotransferase (CGTase) activity was observed when the bacterium was grown in the medium at various initial pH values, containing carbon, nitrogen, phosphorus and mineral salt sources at 50 °C for 24 h in the shake flasks. The optimisation of this growth medium was carried out using response surface methodology. The design contains a total of 32 experimental trials involving 10 star points and 6 replicates at the centre points. The design was employed by selecting sago starch, peptone from casein, K₂HPO₄, CaCl₂ and initial pH as five independent variables in this study. The optimal calculated values of tested variables for maximal production of CGTase were found to be comprised of: sago starch, 16.02 g/l; peptone from casein, 20 g/l; K₂HPO₄, 1.4 g/l; CaCl₂, 0.2 g/l and initial pH, 7.54 with a predicted CGTase activity of 14.20 U/ml. These predicted optimal parameters were tested in the laboratory and the final CGTase activity obtained was very close to the predicted value at 14.80 U/ml.     

Structure and oligomerization of the PilC type IV pilus biogenesis protein from Thermus thermophilus

Type IV pili are expressed from a wide variety of Gram‐negative bacteria and play a major role in host cell adhesion and bacterial motility. PilC is one of at least a dozen different proteins that are implicated in Type IV pilus assembly in Thermus thermophilus and a member of a conserved family of integral inner membrane proteins which are components of the Type II secretion system (GspF) and the archeal flagellum. PilC/GspF family members contain repeats of a conserved helix‐rich domain of around 100 residues in length. Here, we describe the crystal structure of one of these domains, derived from the N‐terminal domain of Thermus thermophilus PilC. The N‐domain forms a dimer, adopting a six helix bundle structure with an up‐down‐up‐down‐up‐down topology. The monomers are related by a rotation of 170°, followed by a translation along the axis of the final α‐helix of approximately one helical turn. This means that the regions of contact on helices 5 and 6 in each monomer are overlapping, but different. Contact between the two monomers is mediated by a network of hydrophobic residues which are highly conserved in PilC homologs from other Gram‐negative bacteria. Site‐directed mutagenesis of residues at the dimer interface resulted in a change in oligomeric state of PilC from tetramers to dimers, providing evidence that this interface is also found in the intact membrane protein and suggesting that it is important to its function. Proteins 2010. © 2010 Wiley‐Liss, Inc.     

Abutilon indicum Exhibits Anxiolytic and Antidepressant Effects in Mice Models

Doklady Biochemistry and Biophysics - Abutilon indicum Linn (A. indicum) is native to tropical and subtropical zones and traditionally used in ulcer, diabetes, piles, jaundice, gonorrhoea and...     

G4 motifs affect origin positioning and efficiency in two vertebrate replicators

DNA replication ensures the accurate duplication of the genome at each cell cycle. It begins at specific sites called replication origins. Genome‐wide studies in vertebrates have recently identified a consensus G‐rich motif potentially able to form G‐quadruplexes (G4) in most replication origins. However, there is no experimental evidence to demonstrate that G4 are actually required for replication initiation. We show here, with two model origins, that G4 motifs are required for replication initiation. Two G4 motifs cooperate in one of our model origins. The other contains only one critical G4, and its orientation determines the precise position of the replication start site. Point mutations affecting the stability of this G4 in vitro also impair origin function. Finally, this G4 is not sufficient for origin activity and must cooperate with a 200‐bp cis‐regulatory element. In conclusion, our study strongly supports the predicted essential role of G4 in replication initiation.     

Self‐Powered Wireless Sensor Node Enabled by a Duck‐Shaped Triboelectric Nanogenerator for Harvesting Water Wave Energy

This paper presents a fully enclosed duck‐shaped triboelectric nanogenerator (TENG) for effectively scavenging energy from random and low‐frequency water waves. The design of the TENG incorporates the freestanding rolling mode and the pitch motion of a duck‐shaped structure generated by incident waves. By investigating the material and structural features, a unit of the TENG device is successfully designed. Furthermore, a hybrid system is constructed using three units of the TENG device. The hybrid system achieves an instantaneous peak current of 65.5 µA with an instantaneous output power density of up to 1.366 W m^?2. Following the design, a fluid–solid interaction analysis is carried out on one duck‐shaped TENG to understand the dynamic behavior, mechanical efficiency, and stability of the device under various water wave conditions. In addition, the hybrid system is experimentally tested to enable a commercial wireless temperature sensor node. In summary, the unique duck‐shaped TENG shows a simple, cost‐effective, environmentally friendly, light‐weight, and highly stable system. The newly designed TENG is promising for building a network of generators to harvest existing blue energy in oceans, lakes, and rivers.     

De novo indol-3-ylmethyl glucosinolate biosynthesis,and not long-distance transport,contributes to defence of Arabidopsis against powdery mildew

Powdery mildew is a fungal disease that affects a wide range of plants and reduces crop yield worldwide. As obligate biotrophs, powdery mildew fungi manipulate living host cells to suppress defence responses and to obtain nutrients. Members of the plant order Brassicales produce indole glucosinolates that effectively protect them from attack by non-adapted fungi. Indol-3-ylmethyl glucosinolate is constitutively produced in the phloem and transported to epidermal cells for storage. Upon attack, indol-3-ylmethyl glucosinolate is activated by CYP81F2 to provide broad-spectrum defence against fungi. How de novo biosynthesis and transport contribute to defence of powdery mildew-attacked epidermal cells is unknown. Bioassays and glucosinolate analysis demonstrate that GTR glucosinolate transporters are not involved in antifungal defence. Using quantitative live-cell imaging of fluorophore-tagged markers, we show that accumulation of the glucosinolate biosynthetic enzymes CYP83B1 and SUR1 is induced in epidermal cells attacked by the non-adapted barley powdery mildew Blumeria graminis f.sp. hordei. By contrast, glucosinolate biosynthesis is attenuated during interaction with the virulent powdery mildew Golovinomyces orontii. Interestingly, SUR1 induction is delayed during the Golovinomyces orontii interaction. We conclude that epidermal de novo synthesis of indol-3-ylmethyl glucosinolate contributes to CYP81F2-mediated broad-spectrum antifungal resistance and that adapted powdery mildews may target this process.     

Clinical significance of plasma MMP‐2 and MMP‐9 levels as biomarkers for tumor expression in breast cancer patients in Egypt

Matrix metallopeptidase 2 (MMP-2) and matrix metallopeptidase 9 (MMP-9) are involved in the breakdown of extracellular matrix in normal physiological processes as well as in disease processes, such as cancer metastasis. We conducted this work to study the role of MMP-2 and MMP-9 in breast cancer by measuring their plasma concentrations before and after surgery. Also, to examine if their levels can reflect the stage of disease and prognosis. Forty-eight breast cancer patients and 13 patients with benign breast diseases were included in the study. MMP-2 and MMP-9 levels were measured by ELISA and semi-quantitative real-time PCR. MMP-2 and MMP-9 levels in plasma were determined by ELISA immediately before surgery and during 6 to 12 months after curative surgery. We observed a significant increase in the level of MMP-9 mRNA expression in breast cancer patients in comparison to their normal breast tissues and to tissues of benign breast disease. In all TNM tumor stages, the plasma levels of MMP-2 and MMP-9 were increased significantly before curative surgery in the studied patients with breast carcinoma and decreased significantly after surgery. Both MMP-2 and MMP-9 may be used as a possible marker for follow-up or as a marker that reflects the response of the disease to treatment.

     

Phenotypic and Molecular Assessment of Wheat Genotypes Tolerant to Leaf Blight,Rust and Blast Diseases

Globally among biotic stresses, diseases like blight, rust and blast constitute prime constraints for reducing wheat productivity especially in Bangladesh. For sustainable productivity, the development of disease-resistant lines and high yielding varieties is vital and necessary. This study was conducted using 122 advanced breeding lines of wheat including 21 varieties developed by Bangladesh Wheat and Maize Research Institute (BAMRI) with aims to identify genotypes having high yield potential and resistance to leaf blight, leaf rust and blast diseases. These genotypes were evaluated for resistance against leaf blight and leaf rust at Dinajpur and wheat blast at Jashore under field condition. Out of 122 genotypes tested, 20 lines were selected as resistant to leaf blight based on the area under the diseases progress curve (AUDPC) under both irrigated timely sown (ITS) and irrigated late sown (ILS) conditions. Forty-two genotypes were found completely free from leaf rust infection, 59 genotypes were identified as resistant, and 13 genotypes were identified as moderately resistant to leaf rust. Eighteen genotypes were immune against wheat blast, 42 genotypes were categorized as resistant, and 26 genotypes were identified as moderately resistant to wheat blast. Molecular data revealed that the 16 genotypes showed a positive 2NS segment among the 18 immune genotypes selected against wheat blast under field conditions. The genotypes BAW 1322, BAW 1295, and BAW 1203 can be used as earlier maturing genotypes and the genotypes BAW 1372, BAW 1373, BAW 1297 and BAW 1364 can be used for lodging tolerant due to short plant height. The genotypes WMRI Gom 1, BAW 1349 and BAW 1350 can be selected for bold grain and the genotypes WMRI Gom 1, BAW 1297, BAW 1377 can be used as high yielder for optimum seeding condition but genotypes BAW 1377 and BAW 1366 can be used for late sown condition. The selected resistant genotypes against specific diseases can be used in the further breeding program to develop wheat varieties having higher disease resistance and yield potential.     

Selection of fatty acid auxotrophs from the oleaginous yeast Cryptococcus curvatus and production of cocoa butter equivalents in batch culture

Summary A search was carried out for mutants, defective in the conversion of stearic acid to oleic acid in effort to improve the quality of lipids produced by Cryptococcus curvatus ATCC, 20509. Mutants were selected as unsaturated fatty acid (Ufa) auxotrophs. After treatment of parent organism with Ethyl methanesulfonate (EMS), 11 oleate-requiring auxotrophs were isolated. Only 3 of them were real unsaturated fatty acid (Ufa) mutants, while the other 8 were designated as fatty acid synthetase (Fas) mutants. The amount of saturated fatty acid (SFA) was about 65.2 % in the lipids extracted from an Ufa mutant named UfaM3 and it was significantly higher than that of the wild-type (WT) (46.6 %) and similar to that of cocoa butter (60.4 %).