Discovery and Characterization of a 5-Hydroxymethylfurfural Oxidase from Methylovorus sp. Strain MP688

In the search for useful and renewable chemical building blocks, 5-hydroxymethylfurfural (HMF) has emerged as a very promising candidate, as it can be prepared from sugars. HMF can be oxidized to 2,5-furandicarboxylic acid (FDCA), which is used as a substitute for petroleum-based terephthalate in polymer production. On the basis of a recently identified bacterial degradation pathway for HMF, candidate genes responsible for selective HMF oxidation have been identified. Heterologous expression of a protein from Methylovorus sp. strain MP688 in Escherichia coli and subsequent enzyme characterization showed that the respective gene indeed encodes an efficient HMF oxidase (HMFO). HMFO is a flavin adenine dinucleotide-containing oxidase and belongs to the glucose-methanol-choline-type flavoprotein oxidase family. Intriguingly, the activity of HMFO is not restricted to HMF, as it is active with a wide range of aromatic primary alcohols and aldehydes. The enzyme was shown to be relatively thermostable and active over a broad pH range. This makes HMFO a promising oxidative biocatalyst that can be used for the production of FDCA from HMF, a reaction involving both alcohol and aldehyde oxidations.

Clinical experience with an epoxy resin adhesive for direct bonding of orthodontic attachments*

KIMAn epoxy resin formulation developed in the Dental Research Unit has been subjected to further clinical trials. Six patients, were used in the study. They were divided into two age groups and all the teeth, including the first molars, bonded. Rubber dam was used to keep the crowns dry during the setting period of the resin. In the younger group the mean failure rates were 27 per cent for anterior teeth and 70 per cent for the first molars. In the older group these rates were 11 per cent for incisors, 11 per cent for canines, 20 per cent for premolars and 20 per cent for first molars. While no figures are available in the literature relating to direct bonding to first molars, the results obtained in this study for the other teeth compare favourably with data previously published by other authors after trials using other types of cement

Origin of Listeria monocytogenes on meat products

Listeria monocytogenes is a relevant food safety hazard in ready to eat products. Inactivation during processing, prevention of recontamination and control of multiplication are the main instruments to secure the safety of meat products. Intensive microbiological monitoring of products and the production environment are valuable tools to assess the level of control in a meat processing plant. During the course of a year all isolates found during hygiene monitoring at a meat processing plant were stored at -70 degrees Celsius. A total of 94 L. monocytogenes isolates have been analyzed by pulsed-field gel electrophoresis (PFGE) and were divided into 30 different types

A single mutation in the core domain of the lac repressor reduces leakiness.

BACKGROUND: The lac operon provides cells with the ability to switch from glucose to lactose metabolism precisely when necessary. This metabolic switch is mediated by the lac repressor (LacI), which in the absence of lactose binds to the operator DNA sequence to inhibit transcription. Allosteric rearrangements triggered by binding of the lactose isomer allolactose to the core domain of the repressor impede DNA binding and lift repression. In Nature, the ability to detect and respond to environmental conditions comes at the cost of the encoded enzymes being constitutively expressed at low levels. The readily-switched regulation provided by LacI has resulted in its widespread use for protein overexpression, and its applications in molecular biology represent early examples of synthetic biology. However, the leakiness of LacI that is essential for the natural function of the lac operon leads to an increased energetic burden, and potentially toxicity, in heterologous protein production. RESULTS: Analysis of the features that confer promiscuity to the inducer-binding site of LacI identified tryptophan 220 as a target for saturation mutagenesis. We found that phenylalanine (similarly to tryptophan) affords a functional repressor that is still responsive to IPTG. Characterisation of the W220F mutant, LacIWF, by measuring the time dependence of GFP production at different IPTG concentrations and at various incubation temperatures showed a 10-fold reduction in leakiness and no decrease in GFP production. Cells harbouring a cytotoxic protein under regulatory control of LacIWF showed no decrease in viability in the early phases of cell growth. Changes in responsiveness to IPTG observed in vivo are supported by the thermal shift assay behaviour of purified LacIWF with IPTG and operator DNA. CONCLUSIONS: In LacI, long-range communications are responsible for the transmission of the signal from the inducer binding site to the DNA binding domain and our results are consistent with the involvement of position 220 in modulating these. The mutation of this single tryptophan residue to phenylalanine generated an enhanced repressor with a 10-fold decrease in leakiness. By minimising the energetic burden and cytotoxicity caused by leakiness, LacIWF constitutes a useful switch for protein overproduction and synthetic biology.RIGHTS : This article is licensed under the BioMed Central licence at http://www.biomedcentral.com/about/license which is similar to the 'Creative Commons Attribution Licence'. In brief you may : copy, distribute, and display the work; make derivative works; or make commercial use of the work - under the following conditions: the original author must be given credit; for any reuse or distribution, it must be made clear to others what the license terms of this work are

Failed resuscitation of a newborn due to congenital tracheal agenesis: a case report

Tracheal agenesis is a rare congenital condition. It usually presents as an unexpected emergency during resuscitation of a newborn in the delivery room. The condition is almost always fatal in the resuscitation phase, but also when the neonate survives the long term prognosis remains poor. We present a case of tracheal agenesis, discuss its presenting symptoms and possibilities for antenatal diagnosis and review the therapeutic options

Water Resources Systems Planning and Management: An Introduction to Methods, Models and Applications

This 2005 version has been superseded by the 2017 edition, available in full here: http://hdl.handle.net/1813/48159Throughout history much of the world has witnessed ever-greater demands for reliable, high-quality and inexpensive water supplies for domestic consumption, agriculture and industry. In recent decades there have also been increasing demands for hydrological regimes that support healthy and diverse ecosystems, provide for water-based recreational activities, reduce if not prevent floods and droughts, and in some cases, provide for the production of hydropower and ensure water levels adequate for ship navigation. Water managers are challenged to meet these multiple and often conflicting demands. At the same time, public stakeholder interest groups have shown an increasing desire to take part in the water resources development and management decision making process. Added to all these management challenges are the uncertainties of natural water supplies and demands due to changes in our climate, changes in people's standards of living, changes in watershed land uses and changes in technology. How can managers develop, or redevelop and restore, and then manage water resources systems - systems ranging from small watersheds to those encompassing large river basins and coastal zones - in a way that meets society's changing objectives and goals? In other words, how can water resources systems become more integrated and sustainable

Factors that Determine the Extent of Business Process Standardization and the Subsequent Effect on Business Performance

Business process standardization is the activity of unifying different variants of a family of business processes. While the positive effects of business process standardization are well-described, it is often undesirable to fully unify different variants due to cultural, legal, or operational reasons. Consequently, a decision has to be made about the extent to which a family of business processes should be standardized. However, little is known about the factors that drive that decision. This paper fills that gap, by presenting factors that drive the extent to which business processes can be standardized, performance properties that are influenced by business process standardization, and relations between these concepts

Evidence for the involvement of tyrosine-69 in the control of stereospecificity of porcine pancreatic phospholipase A2

We have studied the role of Tyr-69 of porcine pancreatic phospholipase A2 in catalysis and substrate binding, using site-directed mutagenesis. A mutant was constructed containing Phe at position 69. Kinetic characterization revealed that the Phe-69 mutant has retained enzymatic activity on monomeric and micellar substrates, and that the mutation has only minor effects on kcat and Km. This shows that Tyr-69 plays no role in the true catalytic events during substrate hydrolysis. In contrast, the mutation has a profound influence on the stereospecificity of the enzyme. Whereas the wild-type phospholipase A2 is only able to catalyse the degradation of sn-3 phospholipids, the Phe-69 mutant hydrolyses both the sn-3 isomers and, at a low (1-2%) rate, the sn-1 isomers. Despite the fact that the stereospecificity of the mutant phospholipase has been altered, Phe-69 phospholipase still requires Ca2+ ions as a cofactor and also retains its specificity for the sn-2 ester bond. Our data suggest that in porcine pancreatic phospholipase A2 the hydroxyl group of Tyr-69 serves to fix and orient the phosphate group of phospholipid monomers by hydrogen bonding. Because no such interaction can occur between the Phe-69 side-chain and the phosphate moiety of the substrate monomer, the mutant enzyme loses part of its stereospecificity but not its positional specificity