title: The dependence of enzyme activity on temperature: determination and validation of parameters

authors: Peterson, Michelle E.

subjects: enzyme activity

abstract: Traditionally, the dependence of enzyme activity on temperature has been described by a model consisting of two processes: the catalytic reaction defined by DGDaggercat, and irreversible inactivation defined by DGDaggerinact. However, such a model does not account for the observed temperature-dependent behaviour of enzymes, and a new model has been developed and validated. This model (the Equilibrium Model) describes a new mechanism by which enzymes lose activity at high temperatures, by including an inactive form of the enzyme (Einact) that is in reversible equilibrium with the active form (Eact); it is the inactive form that undergoes irreversible thermal inactivation to the thermally denatured state. This equilibrium is described by an equilibrium constant whose temperature-dependence is characterized in terms of the enthalpy of the equilibrium, DHeq, and a new thermal parameter, Teq, which is the temperature at which the concentrations of Eact and Einact are equal; Teq may therefore be regarded as the thermal equivalent of Km. Characterization of an enzyme with respect to its temperature-dependent behaviour must therefore include a determination of these intrinsic properties. The Equilibrium Model has major implications for enzymology, biotechnology and understanding the evolution of enzymes. The present study presents a new direct data-fitting method based on fitting progress curves directly to the Equilibrium Model, and assesses the robustness of this procedure and the effect of assay data on the accurate determination of Teq and its associated parameters. It also describes simpler experimental methods for their determination than have been previously available, including those required for the application of the Equilibrium Model to non-ideal enzyme reactions.

text: The dependence of enzyme activity on temperature: determination and validation of parameters Traditionally, the dependence of enzyme activity on temperature has been described by a model consisting of two processes: the catalytic reaction defined by DGDaggercat, and irreversible inactivation defined by DGDaggerinact. However, such a model does not account for the observed temperature-dependent behaviour of enzymes, and a new model has been developed and validated. This model (the Equilibrium Model) describes a new mechanism by which enzymes lose activity at high temperatures, by including an inactive form of the enzyme (Einact) that is in reversible equilibrium with the active form (Eact); it is the inactive form that undergoes irreversible thermal inactivation to the thermally denatured state. This equilibrium is described by an equilibrium constant whose temperature-dependence is characterized in terms of the enthalpy of the equilibrium, DHeq, and a new thermal parameter, Teq, which is the temperature at which the concentrations of Eact and Einact are equal; Teq may therefore be regarded as the thermal equivalent of Km. Characterization of an enzyme with respect to its temperature-dependent behaviour must therefore include a determination of these intrinsic properties. The Equilibrium Model has major implications for enzymology, biotechnology and understanding the evolution of enzymes. The present study presents a new direct data-fitting method based on fitting progress curves directly to the Equilibrium Model, and assesses the robustness of this procedure and the effect of assay data on the accurate determination of Teq and its associated parameters. It also describes simpler experimental methods for their determination than have been previously available, including those required for the application of the Equilibrium Model to non-ideal enzyme reactions.

year: 2007

title: Water relations and carbon dioxide exchange of epiphytic lichens in the Namib fog desert

authors: Lange, Otto L.

subjects: dew

abstract: Although there is only negligible rainfall, frequent nocturnal fog, dew and high air humidity support a luxurious lichen vegetation in the coastal zone of the central NamibDesert (Namibia). In earlier publications, we have studied ecophysiological performance of a series of epilithic and terrestrial lichens. Here, we have extended this work to three epiphytic species (Heterodermia namaquana, Ramalina lacera, and Xanthoria turbinata) that inhabit the sparse perennial shrubs growing in this area. Our intention, monitoring lichen CO₂ exchange, their waterrelations and microclimate conditions, was to determine the functional mechanisms that allow these epiphytes to exist under the special conditions of a fogdesert. Measurements were conducted mainly during the spring season.
The epiphyticlichens showed response patterns very similar to the epilithic and epigaeic species at the same site. Their metabolism was activated through moistening by dew and/or fog during the night and, in the very early morning, they exhibited the typical brief peak of net photosynthesis (NP) between sunrise and desiccation. The thalli were almost completely dry for the remainder of the day. Average duration of the positive NP during the morning peak was about 3 h. Dew condensation, alone, resulted in activation that provided 58–63% of integrated carbon income (ΣNP) as compared to fog (plus dew). In the late afternoon, there was a tendency for hydration to increase again, due to water vapour uptake at higher air humidity, and this allowed on some days a brief additional period of very low rates of photosynthesis shortly before sunset.
Light response of photosynthesis showed “sun-plant” characteristics with saturation around 1000 μmol m⁻² s⁻¹ photosynthetically active photon flux density (PPFD). Light compensation point (LCP) of CO2exchange after sunrise was highly dependent on actual water content (WC) for X. turbinata: at low hydration it was ca. 10 μmol m⁻² s⁻¹ PPFD whilst, at high WC, it was almost 80 μmol m⁻² s⁻¹ PPFD. In contrast, LCP of R. lacera was almost independent of WC. This phenomenon was probably due to differences in thallus structure.
Maximal attained NP and daily ΣNP both showed a saturation-type response to previous maximal nocturnal WC. Neither parameter was increased substantially when higher maximal thallus WCs were produced by experimental moistening in the night. All three species, despite their different morphologies, performed optimally at the highest nocturnal moistening achieved by natural fog and were not able to make use of higher hydration.
The three studied epiphytes were similar in their chlorophyll-related rates of NP. Due to lower chlorophyll content, dry weight and carbon-related NP of X. turbinata was only about one-third of that of the other two species. The average carbon income on days with fog and/or dew hydration during the spring season amounted to 2.4 and 2.1 mgC (gC)⁻¹ day⁻¹ (related to thallus carbon content) for H. namaquana and R. lacera, respectively. This primary production was of similar magnitude to those found for the terrestrial species at the same site.

text: Water relations and carbon dioxide exchange of epiphytic lichens in the Namib fog desert Although there is only negligible rainfall, frequent nocturnal fog, dew and high air humidity support a luxurious lichen vegetation in the coastal zone of the central NamibDesert (Namibia). In earlier publications, we have studied ecophysiological performance of a series of epilithic and terrestrial lichens. Here, we have extended this work to three epiphytic species (Heterodermia namaquana, Ramalina lacera, and Xanthoria turbinata) that inhabit the sparse perennial shrubs growing in this area. Our intention, monitoring lichen CO₂ exchange, their waterrelations and microclimate conditions, was to determine the functional mechanisms that allow these epiphytes to exist under the special conditions of a fogdesert. Measurements were conducted mainly during the spring season.
The epiphyticlichens showed response patterns very similar to the epilithic and epigaeic species at the same site. Their metabolism was activated through moistening by dew and/or fog during the night and, in the very early morning, they exhibited the typical brief peak of net photosynthesis (NP) between sunrise and desiccation. The thalli were almost completely dry for the remainder of the day. Average duration of the positive NP during the morning peak was about 3 h. Dew condensation, alone, resulted in activation that provided 58–63% of integrated carbon income (ΣNP) as compared to fog (plus dew). In the late afternoon, there was a tendency for hydration to increase again, due to water vapour uptake at higher air humidity, and this allowed on some days a brief additional period of very low rates of photosynthesis shortly before sunset.
Light response of photosynthesis showed “sun-plant” characteristics with saturation around 1000 μmol m⁻² s⁻¹ photosynthetically active photon flux density (PPFD). Light compensation point (LCP) of CO2exchange after sunrise was highly dependent on actual water content (WC) for X. turbinata: at low hydration it was ca. 10 μmol m⁻² s⁻¹ PPFD whilst, at high WC, it was almost 80 μmol m⁻² s⁻¹ PPFD. In contrast, LCP of R. lacera was almost independent of WC. This phenomenon was probably due to differences in thallus structure.
Maximal attained NP and daily ΣNP both showed a saturation-type response to previous maximal nocturnal WC. Neither parameter was increased substantially when higher maximal thallus WCs were produced by experimental moistening in the night. All three species, despite their different morphologies, performed optimally at the highest nocturnal moistening achieved by natural fog and were not able to make use of higher hydration.
The three studied epiphytes were similar in their chlorophyll-related rates of NP. Due to lower chlorophyll content, dry weight and carbon-related NP of X. turbinata was only about one-third of that of the other two species. The average carbon income on days with fog and/or dew hydration during the spring season amounted to 2.4 and 2.1 mgC (gC)⁻¹ day⁻¹ (related to thallus carbon content) for H. namaquana and R. lacera, respectively. This primary production was of similar magnitude to those found for the terrestrial species at the same site.

year: 2007

title: Te mātauranga whakatere waka

authors: Tuaupiki, Jackie Whetumarama

abstract: Hei tēnei tuhinga e whai ake nei, ka tirohia te hangarau waka a ngā tūpuna o Te Moana nui a Kiwa i whakawhiti ai rātou ki ō rātou moutere tae atu ki te mātauranga whakatere waka me te arotahi o te titiro ki te whakatinanatanga o aua mātauranga. Ka tāmutu i konā, ka pūhia te titiro i runga i ngā huarere o Poronīhia e kite ai tātou i ngā āhuatanga o Tāwhirimātea i ōna moutere me ngā wero ki te whakatere waka i ērā huarere.

text: Te mātauranga whakatere waka Hei tēnei tuhinga e whai ake nei, ka tirohia te hangarau waka a ngā tūpuna o Te Moana nui a Kiwa i whakawhiti ai rātou ki ō rātou moutere tae atu ki te mātauranga whakatere waka me te arotahi o te titiro ki te whakatinanatanga o aua mātauranga. Ka tāmutu i konā, ka pūhia te titiro i runga i ngā huarere o Poronīhia e kite ai tātou i ngā āhuatanga o Tāwhirimātea i ōna moutere me ngā wero ki te whakatere waka i ērā huarere.

year: 2017