Engineering a Cytidine Aminotransferase for Biocatalytic Production of the Antiviral Molnupiravir

The COVID-19 pandemic highlights the urgent need for cost-effective processes to rapidly manufacture antiviral drugs at scale. Here we report a concise biocatalytic process for Molnupiravir, a nucleoside analogue currently in phase 3 clinical trials as an orally available treatment for SARS-CoV-2. Key to the success of this process was the development of a cytidine aminotransferase for the production of N-hydroxy-cytidine through evolutionary adaption of the hydrolytic enzyme cytidine deaminase. This engineered biocatalyst performs >100,000 turnovers in less than 30 minutes, operates at 180 g/L substrate loading and benefits from in situ crystallization of the N-hydroxy-cytidine product (>90% yield), which can be converted to Molnupiravir by a selective 5’-acylation using Novozym® 435.

Synthesis of ZIF-11 Membranes: The Influence of Preparation Technique and Support Type

Due to its structural features, ZIF-11 is one of the most interesting materials for gas separation applications. Herein, we report a systematic study on the synthesis of ZIF11 as a supported membrane. For this, we adapted optimized conditions for the ZIF-11 powder synthesis, identified in our previous works, to form ZIF layers on symmetric and asymmetric stainless-steel and asymmetric αAl2O3 supports. Different techniques were investigated for the challenging layer formation, namely, in situ crystallization (ISC), multiple in situ crystallization (MISC), and the seeding and secondary growth (SSG) method. It was possible to deposit ZIF11 on different supports by ISC and MISC, although it was difficult to obtain complete layers. SSG, in turn, was more effective in forming dense and well-intergrown ZIF-11 layers. This agrees well with the generally accepted fact that seeding considerably facilitates layer formation. Systematic studies of both individual steps of SSG (seeding and secondary growth) led to a basic understanding of layer formation of ZIF-11 on the different supports. The best membranes prepared by rub seeding and secondary growth achieved Knudsen selectivity. Improved gas separation performance is expected if the formation of defects can be avoided.

Experimental constraints on dacite magma storage beneath Volcán Quizapu, Chile

Volcán Quizapu, Chile, is an under-monitored volcano that was the site of two historical eruptions: an effusive eruption in 1846-1847 and a Plinian eruption in 1932, both of which discharged ∼5 km3 (DRE) of lava and/or tephra. The majority of material erupted in both cases is trachydacite, nearly identical for each event. We present H2O-saturated, phase equilibrium experiments on this end-member dacite magma, using a pumice sample from the 1932 eruption as the main starting material. At an oxygen fugacity (fO2) of ∼NNO+0.2, the phase assemblage of An25-30 plagioclase + amphibole + orthopyroxene, without biotite, is stable at 865±10 °C and 110±20 MPa H2O pressure (PH2O), corresponding to ∼4 km depth. At these conditions, experiments also reproduce the quenched glass composition of the starting pumice. At slightly higher PH2O and below 860 °C, biotite joins the equilibrium assemblage. Because biotite is not part of the observed Quizapu phase assemblage, its presence places an upper limit on PH2O. At the determined storage PH2O of ∼110 MPa, H2O undersaturation of the magma with XH20fluid==0.87 would align Ptotal to mineral-based geobarometry estimates of ∼130 MPa. However, XH20fluid=1< 1 is not required to reproduce the Quizapu dacite phase assemblage and compositions. A second suite of experiments at lower fO2 shows that the stability fields of the hydrous silicates (amphibole and biotite) are significantly restricted at NNO-2 relative to NNO+0.2. Additional observations of Quizapu lava and pumice samples support the existing hypothesis that rapid pre-eruptive heating drove the effusive 1846-1847 eruption, with important refinements. We demonstrate that microlites in the end-member dacite lavas are consistent with in situ crystallization (during ascent), rather than transfer from an andesite. In one end-member dacite lava, newly identified reverse zoning in orthopyroxene and incipient destabilization of amphibole are consistent with small degrees of heating. Our work articulates a clear direction for future Quizapu studies, which are warranted given the active nature of the Cerro Azul-Descabezado Grande volcanic axis.