Engineering the thermotolerant industrial yeast Kluyveromyces marxianus for anaerobic growth

Current large-scale, anaerobic industrial processes for ethanol production from renewable carbohydrates predominantly rely on the mesophilic yeast Saccharomyces cerevisiae. Use of thermotolerant, facultatively fermentative yeasts such as Kluyveromyces marxianus could confer significant economic benefits. However, in contrast to S. cerevisiae, these yeasts cannot grow in the absence of oxygen. Response of K. marxianus and S. cerevisiae to different oxygen-limitation regimes were analyzed in chemostats. Genome and transcriptome analysis, physiological responses to sterol supplementation and sterol-uptake measurements identified absence of a functional sterol-uptake mechanism as a key factor underlying the oxygen requirement of K. marxianus. Heterologous expression of a squalene-tetrahymanol cyclase enabled oxygen-independent synthesis of the sterol surrogate tetrahymanol in K. marxianus. After a brief adaptation under oxygen-limited conditions, tetrahymanol-expressing K. marxianus strains grew anaerobically on glucose at temperatures of up to 45 °C. These results open up new directions in the development of thermotolerant yeast strains for anaerobic industrial applications.

N-hydroxyphthalimidyl diazoacetate (NHPI-DA): a modular methylene linchpin for the C–H alkylation of indoles

Despite the extensive studies on the reactions between conventional diazocompounds and indoles, these are still limited by the independent synthesis of the carbene precursors, the specific catalysts, and the required...

A hitherto unreported impurity in Terazosin – elucidation of the structure, synthesis and cytotoxicity

<p>Analysis of several batches of the a₁–adrenergic blocking agent terazosin being used as a medication for treating benign prostatic hyperplasia and hypertension revealed the presence of a hitherto not reported impurity. The latter was isolated, and its structure was elucidated from NMR and Mass Spectrometry (MS) data and unambiguously confirmed by independent synthesis. This contamination, represented in 1-[4-(amino-6,7-dimethoxyquinazolin-2-yl)-piperazin-1-yl]-pentane-1,2-dione <strong>2 </strong>is likely to occur as the product of a side-reaction in the catalytic hydrogenation step during the synthesis of the drug. Biological screening showed this compound as not cytotoxic for several human tumor cell lines and non-malignant fibroblasts.<strong></strong></p>

ITPK1 mediates the lipid-independent synthesis of inositol phosphates controlled by metabolism

Inositol phosphates (IPs) comprise a network of phosphorylated molecules that play multiple signaling roles in eukaryotes. IPs synthesis is believed to originate with IP3 generated from PIP2 by phospholipase C (PLC). Here, we report that in mammalian cells PLC-generated IPs are rapidly recycled to inositol, and uncover the enzymology behind an alternative “soluble” route to synthesis of IPs. Inositol tetrakisphosphate 1-kinase 1 (ITPK1)—found in Asgard archaea, social amoeba, plants, and animals—phosphorylates I(3)P1 originating from glucose-6-phosphate, and I(1)P1 generated from sphingolipids, to enable synthesis of IP6. We also found using PAGE mass assay that metabolic blockage by phosphate starvation surprisingly increased IP6 levels in a ITPK1-dependent manner, establishing a route to IP6 controlled by cellular metabolic status, that is not detectable by traditional [3H]-inositol labeling. The presence of ITPK1 in archaeal clades thought to define eukaryogenesis indicates that IPs had functional roles before the appearance of the eukaryote.

The Biology of Viruses

In order to be classified as a virus, certain criteria have to be fulfilled. Viruses must ● Be only capable of growth and multiplication within living cells, i.e. obligate intracellular parasite. Host cells could include humans, animals, insects, plants, protozoa, or even bacteria. ● Have a nucleic acid genome (either RNA or DNA, but not both) surrounded by a protein coat (capsid). ● Have no semipermeable membrane, though some have an envelope formed of phospholipids and proteins. ● Be inert outside of the host cell. Enveloped viruses are susceptible to inactivation by organic solvents such as alcohol. ● Perform replication by independent synthesis of components followed by assembly (c.f. binary fission in bacteria). Viruses are considered as a bundle of genetic programmes encoded in nucleic acids and packaged with a capsid +/ - envelope protein, which can be activated on entry into a host cell (compare this with computer viruses packaged in an enticing way in order to infect and take over control of your PC). Although they share some similarities in their properties, mycoplasma and chlamydia are true bacteria. The virion (assembled infectious particle) consists of viral nucleic acid and capsid. The nucleic acid of a virus can either be ribonucleic acid (RNA) or deoxyribonucleic acid (DNA), and the amount of genetic material varies widely, with some viruses able to encode a few proteins and others having genetic material that encodes hundreds of proteins. In association with the nucleic acid there may be non- structural viral proteins, such as a viral polymerase. The nucleic acid and non- structural proteins are protected by a surrounding layer of capsid proteins. The capsid includes proteins which can attach to host cell receptors. The proteins and the cell receptors to which they bind determine a virus’ tropism, i.e., the ability to bind to and enter different cell types. The term nucleocapsid refers to the nucleic acid core surrounded by capsid protein. Some viruses also have an envelope made up of phospholipids and proteins surrounding the nucleocapsid. This envelope can be formed by the host cell membrane during the process of a virus budding from a cell during replication.

Benzothiazole Nickelation: An Obstacle to the Catalytic Arylation of Azoles by Cyclopentadienyl Nickel N-Heterocyclic Carbene Complexes

NiCp†L(NHC)](+) complexes (Cp† = Cp (η5-C5H5), Cp* (η5-C5Me5); NHC = N-heterocyclic carbene; L = Cl− or NCMe) have been tested as pre-catalysts for the direct arylation of benzothiazole in the presence of an alkoxide. Only the pentamethylcyclopentadienyl derivative, [NiCp*Cl(IMes)] (IMes = 1,3-bis(2,4,6-trimethylphenylimidazol-2-ylidene), enabled low conversion to the desired coupling product with phenyl iodide as the electrophilic coupling partner. In contrast, all cyclopentadienyl complexes proved to be inactive. 1H NMR studies of the “catalytic” reaction mixtures demonstrate that they cleanly convert to an unreactive C(2)-benzothiazolyl derivative, whose identity has been confirmed by an independent synthesis and characterization. The latter constitutes a potential energy well that quenches all further reactivity, and provides a rare example of C(2)-metallated azolyl complex.

Substrate-independent and catalyst-free synthesis of magnesium nanowires

We report a catalyst free and substrate independent synthesis of magnesium nanowires using a simple thermal evaporation method.

Peter of Auvergne (d. 1304)

Peter of Auvergne, a thirteenth-century Parisian master, wrote extensively on logic, natural philosophy and theology. His thought progresses from modism in logic to an independent synthesis of Aristotelian philosophy along the lines begun by Thomas Aquinas, culminating in a theology reconciling the ideas of his teachers Henry of Ghent and Godfrey of Fontaines. His reputation has been based largely on his association with Aquinas, but recent investigations have shown the independence of his thought.