title: Tests of predictions made by the Equilibrium Model for the effect of temperature on enzyme activity

authors: Oudshoorn, Matthew Leslie

subjects: enzymes

abstract: The Classical Model describing the effects of temperature on enzyme activity consists of two processes: the catalytic reaction defined by ΔG cat and irreversible inactivation defined by ΔG inact, this model however, does not account for the observed temperature- dependant behaviour of enzymes. The recent development of the Equilibrium Model is governed not only by ΔG cat and ΔG inact but also by two new intrinsic parameters ΔHeq and Teq, which describe the enthalpy and the temperature of the midpoint, respectively, of a active and reversibly inactive enzyme transition. Teq is central to the physiological adaptation of an enzyme to its environmental temperature and links the molecular, physiological and environmental aspects of life to temperature in a way that has not been previously possible. The Equilibrium Model is therefore a more complete and accurate description of the effects of temperature on enzymes, it has provided new tools for describing and investigating enzyme thermal adaptation and possibly new biotechnological tools.

The effects of the incorporating in the new Model of the parameters Teq and ΔH eq yield major differences from the Classical Model, with simulated data calculated according to the Equilibrium Model fitting well to experimental data and showing an initial rate temperature optimum that is independent of assay duration. Simulated data simulated according to the Classical Model can not be fitted to experimental data. All enzymes so far studied (gt30) display behaviour predicted by the Equilibrium Model.

The research described here has set out to: experimentally test observations made by Eisenthal et al., on the basis of enzyme reactor data simulated according to the Equilibrium Model; to test the Equilibrium Model using an unusual (rapidly renaturable) enzyme, RNAase; and to test the proposed molecular basis of the Equilibrium Model by examining the effect of a change at the enzymes active site.

The experimental results gathered here on the effect of time and temperature on enzyme reactor output confirm the predictions made by Eisenthal et al. (2006) and indicate that the Equilibrium Model can be a useful aid in predicting reactor performance. The Equilibrium Model depends upon the acquisition of data on the variation of the Vmax of an enzyme with time and temperature, and the non-ideal behaviour of RNase A made it impossible to collect such data for this enzyme, as a result the Equilibrium Model could not be applied. The disulfide bond within the active site cleft of A.k 1 protease was cleaved as a probe of the mechanism of the Equilibrium Model, which is proposed to arise from molecular changes at the enzymes active site. Support for the proposed mechanism was gained through the comparison of experimentally determined temperature dependence of the native and reduced forms of the enzyme and application of this data to the Equilibrium Model.

text: Tests of predictions made by the Equilibrium Model for the effect of temperature on enzyme activity The Classical Model describing the effects of temperature on enzyme activity consists of two processes: the catalytic reaction defined by ΔG cat and irreversible inactivation defined by ΔG inact, this model however, does not account for the observed temperature- dependant behaviour of enzymes. The recent development of the Equilibrium Model is governed not only by ΔG cat and ΔG inact but also by two new intrinsic parameters ΔHeq and Teq, which describe the enthalpy and the temperature of the midpoint, respectively, of a active and reversibly inactive enzyme transition. Teq is central to the physiological adaptation of an enzyme to its environmental temperature and links the molecular, physiological and environmental aspects of life to temperature in a way that has not been previously possible. The Equilibrium Model is therefore a more complete and accurate description of the effects of temperature on enzymes, it has provided new tools for describing and investigating enzyme thermal adaptation and possibly new biotechnological tools.

The effects of the incorporating in the new Model of the parameters Teq and ΔH eq yield major differences from the Classical Model, with simulated data calculated according to the Equilibrium Model fitting well to experimental data and showing an initial rate temperature optimum that is independent of assay duration. Simulated data simulated according to the Classical Model can not be fitted to experimental data. All enzymes so far studied (gt30) display behaviour predicted by the Equilibrium Model.

The research described here has set out to: experimentally test observations made by Eisenthal et al., on the basis of enzyme reactor data simulated according to the Equilibrium Model; to test the Equilibrium Model using an unusual (rapidly renaturable) enzyme, RNAase; and to test the proposed molecular basis of the Equilibrium Model by examining the effect of a change at the enzymes active site.

The experimental results gathered here on the effect of time and temperature on enzyme reactor output confirm the predictions made by Eisenthal et al. (2006) and indicate that the Equilibrium Model can be a useful aid in predicting reactor performance. The Equilibrium Model depends upon the acquisition of data on the variation of the Vmax of an enzyme with time and temperature, and the non-ideal behaviour of RNase A made it impossible to collect such data for this enzyme, as a result the Equilibrium Model could not be applied. The disulfide bond within the active site cleft of A.k 1 protease was cleaved as a probe of the mechanism of the Equilibrium Model, which is proposed to arise from molecular changes at the enzymes active site. Support for the proposed mechanism was gained through the comparison of experimentally determined temperature dependence of the native and reduced forms of the enzyme and application of this data to the Equilibrium Model.

year: 2008

title: Religion in the Te Aroha district

authors: Hart, Philip

abstract: The Christians of the district were split into a variety of denominations, though only a few had strong followings. Opinions varied about the religious zeal of miners, and although the first services to be held after the initial rush to the new goldfield were well attended, this may at least in part have been out of curiosity. 
The Protestant denominations were notable for ecumenical activities, notably in the temperance movement, running societies for the young, and evangelical crusades. They encouraged religious education, and the only Sunday School, run by the Wesleyans, was attended by children from other denominations. Special meetings and entertainments held by Protestants to attract converts and to strengthen the faith of believers were regularly held and attended by many, even some Catholics. In general, relations between Protestants and Catholics were warm, and there were some ‘mixed marriages’. 
Details are given of the development of the Anglican, Presbyterian, Methodist, and Catholic churches, covering the erection and expansion of church buildings, their clergy, the expansion of their congregations, and special activities, both social and religious – the former because of the constant need to raise funds. It is impossible to prove the extent and depth of belief, but the moral behaviour of some residents meant that continual efforts were seen as necessary to get them to change their ways.

text: Religion in the Te Aroha district The Christians of the district were split into a variety of denominations, though only a few had strong followings. Opinions varied about the religious zeal of miners, and although the first services to be held after the initial rush to the new goldfield were well attended, this may at least in part have been out of curiosity. 
The Protestant denominations were notable for ecumenical activities, notably in the temperance movement, running societies for the young, and evangelical crusades. They encouraged religious education, and the only Sunday School, run by the Wesleyans, was attended by children from other denominations. Special meetings and entertainments held by Protestants to attract converts and to strengthen the faith of believers were regularly held and attended by many, even some Catholics. In general, relations between Protestants and Catholics were warm, and there were some ‘mixed marriages’. 
Details are given of the development of the Anglican, Presbyterian, Methodist, and Catholic churches, covering the erection and expansion of church buildings, their clergy, the expansion of their congregations, and special activities, both social and religious – the former because of the constant need to raise funds. It is impossible to prove the extent and depth of belief, but the moral behaviour of some residents meant that continual efforts were seen as necessary to get them to change their ways.

year: 2016

title: Māori wards in Tauranga Moana and Aotearoa: Liminal local government democracy

authors: Des Forges, Sandra

subjects: Māori wards

abstract: This thesis focuses on Māori representation within the highly contested arena of local government democracy in Aotearoa New Zealand. Limitations of the current Eurocentric model of local government democracy are considered at various spatial scales. Legislative changes to enable councils to establish Māori wards were launched in the early 2000s. Māori wards are defined locations where those on the Māori electoral roll vote for Māori ward candidates, resulting in dedicated Māori representation at elected member level. Until 2021 when the legislation was amended to remove the ability for public referenda to challenge council decisions on establishing Māori wards, however, little change occurred. Since then, 35 councils have established Māori wards for the 2022 local body elections.
The research asks how, and in what ways, do Māori wards decolonise local government and encourage greater Māori representation? To explore this question, the empirical chapters are framed around three key aspects of liminality; the in-between positioning of Māori wards reflecting a time of change; this liminal space being a time where unease and discomfort is experienced by some as Indigenous disparity is addressed; and the threshold positioning of Māori wards as a place of opportunity and creativity, where new ideas and practices may be considered. Geographical concepts of (un)belonging, exclusion and deep colonising reveal challenges for Māori participation at the level of local government decision-making. 
The research uses an Indigenous methodological framework, Te Ara Tika, based on a framework of Kaupapa Māori, and developed specifically for non-Māori researchers. Māori wards have been widely debated in the media, particularly at the time of changes to the Māori ward legislation in 2021. Thematic analysis of 122 media representations of the Māori ward debate constitutes part of the empirical evidence. Fourteen individual interviews with local government representatives, including elected Council members from Tauranga Moana and members of Te Rangapū Mana Whenua o Tauranga Moana Partnership, were conducted. Additionally, participant observations took place at three public meetings on Māori wards.
The way the current form of local government representative democracy impacts Māori representation is identified. The research demonstrates how local government continues to act as a tool of colonialism by privileging Western worldviews, institutions, and systems. The research finds that Māori wards are liminal democratic options that are both decolonising and deep colonising. Evidence shows that the liminal space of Māori wards is an opportunity to consider options for a way forward to reimagine local government as a place of belonging underpinned by Te Tiriti o Waitangi and Te Ao Māori principles. 
The research builds on decolonisation literatures that seek to unsettle the hegemony of Eurocentric institutions and systems within colonised countries such as Aotearoa. Providing a critical spatial perspective on the intersection of democracy, colonisation and Indigeneity, this thesis advances decolonising geographical knowledges. In particular, this research advances debates about democratic processes, exposing ways in which colonially-based local authority democratic mechanisms contribute to under-representation issues. A reimagined local government allows current hegemonic approaches to be rethought and provides insights for a shift towards genuine decolonising processes.

text: Māori wards in Tauranga Moana and Aotearoa: Liminal local government democracy This thesis focuses on Māori representation within the highly contested arena of local government democracy in Aotearoa New Zealand. Limitations of the current Eurocentric model of local government democracy are considered at various spatial scales. Legislative changes to enable councils to establish Māori wards were launched in the early 2000s. Māori wards are defined locations where those on the Māori electoral roll vote for Māori ward candidates, resulting in dedicated Māori representation at elected member level. Until 2021 when the legislation was amended to remove the ability for public referenda to challenge council decisions on establishing Māori wards, however, little change occurred. Since then, 35 councils have established Māori wards for the 2022 local body elections.
The research asks how, and in what ways, do Māori wards decolonise local government and encourage greater Māori representation? To explore this question, the empirical chapters are framed around three key aspects of liminality; the in-between positioning of Māori wards reflecting a time of change; this liminal space being a time where unease and discomfort is experienced by some as Indigenous disparity is addressed; and the threshold positioning of Māori wards as a place of opportunity and creativity, where new ideas and practices may be considered. Geographical concepts of (un)belonging, exclusion and deep colonising reveal challenges for Māori participation at the level of local government decision-making. 
The research uses an Indigenous methodological framework, Te Ara Tika, based on a framework of Kaupapa Māori, and developed specifically for non-Māori researchers. Māori wards have been widely debated in the media, particularly at the time of changes to the Māori ward legislation in 2021. Thematic analysis of 122 media representations of the Māori ward debate constitutes part of the empirical evidence. Fourteen individual interviews with local government representatives, including elected Council members from Tauranga Moana and members of Te Rangapū Mana Whenua o Tauranga Moana Partnership, were conducted. Additionally, participant observations took place at three public meetings on Māori wards.
The way the current form of local government representative democracy impacts Māori representation is identified. The research demonstrates how local government continues to act as a tool of colonialism by privileging Western worldviews, institutions, and systems. The research finds that Māori wards are liminal democratic options that are both decolonising and deep colonising. Evidence shows that the liminal space of Māori wards is an opportunity to consider options for a way forward to reimagine local government as a place of belonging underpinned by Te Tiriti o Waitangi and Te Ao Māori principles. 
The research builds on decolonisation literatures that seek to unsettle the hegemony of Eurocentric institutions and systems within colonised countries such as Aotearoa. Providing a critical spatial perspective on the intersection of democracy, colonisation and Indigeneity, this thesis advances decolonising geographical knowledges. In particular, this research advances debates about democratic processes, exposing ways in which colonially-based local authority democratic mechanisms contribute to under-representation issues. A reimagined local government allows current hegemonic approaches to be rethought and provides insights for a shift towards genuine decolonising processes.

year: 2023