Metabolic Deficiences Revealed in the Biotechnologically Important Model Bacterium Escherichia coli BL21(DE3)

The Escherichia coli B strain BL21(DE3) has had a profound impact on biotechnology through its use in the production of recombinant proteins. Little is understood, however, regarding the physiology of this important E. coli strain. We show here that BL21(DE3) totally lacks activity of the four [NiFe]-hydrogenases, the three molybdenum- and selenium-containing formate dehydrogenases and molybdenum-dependent nitrate reductase. Nevertheless, all of the structural genes necessary for the synthesis of the respective anaerobic metalloenzymes are present in the genome. However, the genes encoding the high-affinity molybdate transport system and the molybdenum-responsive transcriptional regulator ModE are absent from the genome. Moreover, BL21(DE3) has a nonsense mutation in the gene encoding the global oxygen-responsive transcriptional regulator FNR. The activities of the two hydrogen-oxidizing hydrogenases, therefore, could be restored to BL21(DE3) by supplementing the growth medium with high concentrations of Ni2+ (Ni2+-transport is FNR-dependent) or by introducing a wild-type copy of the fnr gene. Only combined addition of plasmid-encoded fnr and high concentrations of MoO42− ions could restore hydrogen production to BL21(DE3); however, to only 25–30% of a K-12 wildtype. We could show that limited hydrogen production from the enzyme complex responsible for formate-dependent hydrogen evolution was due solely to reduced activity of the formate dehydrogenase (FDH-H), not the hydrogenase component. The activity of the FNR-dependent formate dehydrogenase, FDH-N, could not be restored, even when the fnr gene and MoO42− were supplied; however, nitrate reductase activity could be recovered by combined addition of MoO42− and the fnr gene. This suggested that a further component specific for biosynthesis or activity of formate dehydrogenases H and N was missing. Re-introduction of the gene encoding ModE could only partially restore the activities of both enzymes. Taken together these results demonstrate that BL21(DE3) has major defects in anaerobic metabolism, metal ion transport and metalloprotein biosynthesis

The influence of UVB-radiation on light-dependent photosynthetic performance in Sanionia unicata (Hedw.) Loeske in Antarctica

Photosynthetic activity of the moss Sanionia uncinata (Hedw.) Loeske was investigated on Léonie Island (67°35'S, 68°20'W, Antarctic Peninsula) in response to short-term changes of UV-B radiation. The UV-environment of natural mat formations dominated by S. uncinata was altered using filter screens. Two filter experiments were conducted in the Antarctic summers 1998 and 1999. A third filter experiment was conducted during springtime ozone depletion in October 1998. Photosynthetic activity of S. uncinata was mainly determined by photosynthetically active photon flux density (PPFD). Light response of relative electron transport rate through photosystem II (rel ETR=F/Fm'×PPFD) remained unaffected by ambient summer levels of UV-B radiation. The same was found for net photosynthesis and dark respiration. In October 1998, S. uncinata was mainly metabolically inactive due to low temperatures. No significant levels of DNA-damage measured as cyclobutane pyrimidine dimers (CPDs) were induced by ambient summer levels of UV-B. Artificially enhanced UV-B radiation supplying a Setlow-DNA-dose of 8.7 kJ m–2 day–1 UV-B led to formation of 7±3 CPD (106 nucleotides)–1. It is concluded that current ambient summer levels of UV-B radiation do not affect photosynthetic activity in S. uncinata.

The influence of UV-B radiation on light-dependent photosynthetic performance in Sanionia uncinata (Hedw.) Loeske in Antarctica

Photosynthetic activity of the moss Sanionia uncinata (Hedw.) Loeske was investigated on Léonie Island (67°35'S, 68°20'W, Antarctic Peninsula) in response to short-term changes of UV-B radiation. The UV-environment of natural mat formations dominated by S. uncinata was altered using filter screens. Two filter experiments were conducted in the Antarctic summers 1998 and 1999. A third filter experiment was conducted during springtime ozone depletion in October 1998. Photosynthetic activity of S. uncinata was mainly determined by photosynthetically active photon flux density (PPFD). Light response of relative electron transport rate through photosystem II (rel ETR=DeltaF/Fm'×PPFD) remained unaffected by ambient summer levels of UV-B radiation. The same was found for net photosynthesis and dark respiration. In October 1998, S. uncinata was mainly metabolically inactive due to low temperatures. No significant levels of DNA-damage measured as cyclobutane pyrimidine dimers (CPDs) were induced by ambient summer levels of UV-B. Artificially enhanced UV-B radiation supplying a Setlow-DNA-dose of 8.7 kJ m–2 day–1 UV-B led to formation of 7±3 CPD (106 nucleotides)–1. It is concluded that current ambient summer levels of UV-B radiation do not affect photosynthetic activity in S. uncinata

Estimation of the annual primary production of the lichen Cetrariella delisei in a glacier foreland in the High Arctic, Ny-Ølesund, Svalbard

The fruticose lichen Cetrariella delisei is among the dominant lichen species in the deglaciated High Arctic areas of Svalbard. As part of a study of carbon cycling in the High Arctic, we aimed to estimate the primary production of lichen in a deglaciated area in Ny-Ålesund, Svalbard (79° N), by examining the effects of abiotic factors on the net photosynthesis (Pn) and dark respiration (R) rates of C. delisei. Experiments were conducted in the snow-free season of 2000 using an open-fl ow gas exchange system with an infrared gas analyser. Positive photosynthetic activities were observed on rainy days or soon after rainfall when the thallus water content was high, whereas photosynthetic activities dropped below the detectable limit on clear days because of the low thallus water content. Under a suffi ciently high thallus water content and close to light saturation, Pn was nearly constant over a wide temperature range of 4 - 20 °C, while R increased with increasing temperature. We constructed a model for estimating the net primary production (NPP) of lichen based on the relationships between abiotic factors and the CO2 exchange rate. The mean, minimum and maximum NPP values in the snow-free season, estimated using meteorological data obtained from 1995–2003, were 5.1, 1.0 and 8.4 g dry wt. m–2 snow-free season–1, respectively. These results suggest that NPP is highly variable and the contribution of lichen to carbon input is small compared with that of vascular plants and mosses in the study site