Mutants of Rhizobium japonicum with Increased Hydrogenase Activity

Some strains of Rhizobium japonicum can use hydrogen as an energy source for growth under microaerophilic conditions. Mutant strains have been selected that use hydrogen in the presence of high partial pressures of oxygen. The mutants contain more hydrogenase than the parent strain, both as free-living cells and as bacteroids in nitrogen-fixing soybean root nodules.     

Second round of an interlaboratory comparison of SARS-CoV2 molecular detection assays used by 45 veterinary diagnostic laboratories in the United States

The COVID-19 pandemic presents a continued public health challenge. Veterinary diagnostic laboratories in the United States use RT-rtPCR for animal testing, and many laboratories are certified for testing human samples; hence, ensuring that laboratories have sensitive and specific SARS-CoV2 testing methods is a critical component of the pandemic response. In 2020, the FDA Veterinary Laboratory Investigation and Response Network (Vet-LIRN) led an interlaboratory comparison (ILC1) to help laboratories evaluate their existing RT-rtPCR methods for detecting SARS-CoV2. All participating laboratories were able to detect the viral RNA spiked in buffer and PrimeStore molecular transport medium (MTM). With ILC2, Vet-LIRN extended ILC1 by evaluating analytical sensitivity and specificity of the methods used by participating laboratories to detect 3 SARS-CoV2 variants (B.1; B.1.1.7 [Alpha]; B.1.351 [Beta]) at various copy levels. We analyzed 57 sets of results from 45 laboratories qualitatively and quantitatively according to the principles of ISO 16140-2:2016. More than 95% of analysts detected the SARS-CoV2 RNA in MTM at ≥500 copies for all 3 variants. In addition, for nucleocapsid markers N1 and N2, 81% and 92% of the analysts detected ≤20 copies in the assays, respectively. The analytical specificity of the evaluated methods was >99%. Participating laboratories were able to assess their current method performance, identify possible limitations, and recognize method strengths as part of a continuous learning environment to support the critical need for the reliable diagnosis of COVID-19 in potentially infected animals and humans.     

Volatile compounds of selected strawberry varieties analyzed by purge-and-trap headspace GC-MS

Six strawberry (Fragaria x ananassa Duch.) varieties, Senga Sengana, Jonsok, Polka, Korona, Bounty, and Honeoye, were studied in relation to composition of their volatile headspace compounds collected by dynamic purge-and-trap concentration. Also, the influence of cultivation technique on the volatile compounds was tested by comparing conventionally and organically cultivated strawberry varieties. In addition, the effect of geographical origin on strawberry volatiles was investigated. The emphasis of this study was lay in the industrial utilization of strawberry. A total of 52 different volatile compounds were determined from frozen strawberries grown in 1997 and 1998. In addition to individual compounds, total areas of chromatogram profiles were compared. The major esters were found to be methyl butanoate, ethyl butanoate, methyl hexanoate, and ethyl hexanoate. With the application of principal component analysis and analysis of variance to the data matrix, the varieties were classified into three groups. Var. Honeoye was most different from the others. Organic cultivation had no effect on strawberry volatiles.     

The crystal structure of a mammalian fatty acid synthase

Mammalian fatty acid synthase is a large multienzyme that catalyzes all steps of fatty acid synthesis. We have determined its crystal structure at 3.2 angstrom resolution covering five catalytic domains, whereas the flexibly tethered terminal acyl carrier protein and thioesterase domains remain unresolved. The structure reveals a complex architecture of alternating linkers and enzymatic domains. Substrate shuttling is facilitated by flexible tethering of the acyl carrier protein domain and by the limited contact between the condensing and modifying portions of the multienzyme, which are mainly connected by linkers rather than direct interaction. The structure identifies two additional nonenzymatic domains: (i) a pseudo-ketoreductase and (ii) a peripheral pseudo-methyltransferase that is probably a remnant of an ancestral methyltransferase domain maintained in some related polyketide synthases. The structural comparison of mammalian fatty acid synthase with modular polyketide synthases shows how their segmental construction allows the variation of domain composition to achieve diverse product synthesis.     

C and N dynamics of a range of biogas slurries as a function of application rate and soil texture: a laboratory experiment

Digestates vary in composition and studies regarding their impact on C and N dynamics in soils are scarce. The objective was to analyse the C and N dynamics of digestates originating from various substrates applied to a sandy Cambisol and a silty Anthrosol. In three laboratory experiments (4–6 weeks), the effects of digestate properties, N rate and water content were tested. Averaged over both soils, 21% of the C supplied was emitted as CO₂. Potential NH₃ emissions during the first week ranged between 6% and 12% of NH₄⁺ present in the digestates. The emission factors in the sandy Cambisol were on average 1.2 and 2 times higher for CO₂ and potential NH₃, respectively, compared to the silty Anthrosol. Similarly, net nitrogen mineralization in the sandy Cambisol was approximately twice the N mineralized in the silty Anthrosol. Net nitrification was not influenced by soil texture or different digestates, but increased with increasing application rates and had highest values at 75% of water holding capacity. Our results indicate that the type of substrate input for anaerobic digestion influences the properties of the digestate and therefore the dynamics of C and N. However, soil texture can affect these dynamics markedly.