title: Some characteristics of a proteinase from a thermophilic Bacillus sp. expressed in Escherichia coli: comparison with the native enzyme and its processing in E. coli and in vitro.

authors: Peek, Keith

subjects: biology

abstract: Proteinase Ak.1 was produced during the stationary phase of Bacillus sp. Ak.1 cultures. It is a serine proteinase with a pI of 4.0, and the molecular mass was estimated to be 36.9 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme was stable at 60 and 70 degrees C, with half-lives of 13 h and 19 min at 80 and 90 degrees C, respectively. Maximum proteolytic activity was observed at pH 7.5 with azocasein as a substrate, and the enzyme also cleaved the endoproteinase substrate Suc-Ala-Ala-Pro-Phe-NH-Np (succinyl-alanyl-alanyl-prolyl-phenylalanine p-nitroanalide). Major cleavage sites of the insulin B chain were identified as Leu-15-Tyr-16, Gln-4-His-5, and Glu-13-Ala-14. The proteinase gene was cloned in Escherichia coli, and expression of the active enzyme was detected in the extracellular medium at 75 degrees C. The enzyme is expressed in E. coli as an inactive proproteinase at 37 degrees C and is converted to the mature enzyme by heating the cell-free media to 60 degrees C or above. The proproteinase was purified to homogeneity and had a pI of 4.3 and a molecular mass of 45 kDa. The NH2-terminal sequence was Ala-Ser-Asn-Asp-Gly-Val-Glu-, showing the exact signal peptide cleavage point. Heating the proenzyme resulted in the production of active proteinase with an NH2-terminal sequence identical to that of the native enzyme. The characteristics of the cloned proteinase were identical to those of the native enzyme.

text: Some characteristics of a proteinase from a thermophilic Bacillus sp. expressed in Escherichia coli: comparison with the native enzyme and its processing in E. coli and in vitro. Proteinase Ak.1 was produced during the stationary phase of Bacillus sp. Ak.1 cultures. It is a serine proteinase with a pI of 4.0, and the molecular mass was estimated to be 36.9 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme was stable at 60 and 70 degrees C, with half-lives of 13 h and 19 min at 80 and 90 degrees C, respectively. Maximum proteolytic activity was observed at pH 7.5 with azocasein as a substrate, and the enzyme also cleaved the endoproteinase substrate Suc-Ala-Ala-Pro-Phe-NH-Np (succinyl-alanyl-alanyl-prolyl-phenylalanine p-nitroanalide). Major cleavage sites of the insulin B chain were identified as Leu-15-Tyr-16, Gln-4-His-5, and Glu-13-Ala-14. The proteinase gene was cloned in Escherichia coli, and expression of the active enzyme was detected in the extracellular medium at 75 degrees C. The enzyme is expressed in E. coli as an inactive proproteinase at 37 degrees C and is converted to the mature enzyme by heating the cell-free media to 60 degrees C or above. The proproteinase was purified to homogeneity and had a pI of 4.3 and a molecular mass of 45 kDa. The NH2-terminal sequence was Ala-Ser-Asn-Asp-Gly-Val-Glu-, showing the exact signal peptide cleavage point. Heating the proenzyme resulted in the production of active proteinase with an NH2-terminal sequence identical to that of the native enzyme. The characteristics of the cloned proteinase were identical to those of the native enzyme.

year: 1993

title: Learning Through Utility Optimization in Regression Tasks

authors: Branco, Paula

subjects: Science & Technology

abstract: Accounting for misclassification costs is important in many practical applications of machine learning, and cost-sensitive techniques for classification have been studied extensively. Utility-based learning provides a generalization of purely cost-based approaches that considers both costs and benefits, enabling application to domains with complex cost-benefit settings. However, there is little work on utility- or cost-based learning for regression. In this paper, we formally define the problem of utility-based regression and propose a strategy for maximizing the utility of regression models. We verify our findings in a large set of experiments that show the advantage of our proposal in a diverse set of domains, learning algorithms and cost/benefit settings.

text: Learning Through Utility Optimization in Regression Tasks Accounting for misclassification costs is important in many practical applications of machine learning, and cost-sensitive techniques for classification have been studied extensively. Utility-based learning provides a generalization of purely cost-based approaches that considers both costs and benefits, enabling application to domains with complex cost-benefit settings. However, there is little work on utility- or cost-based learning for regression. In this paper, we formally define the problem of utility-based regression and propose a strategy for maximizing the utility of regression models. We verify our findings in a large set of experiments that show the advantage of our proposal in a diverse set of domains, learning algorithms and cost/benefit settings.

year: 2017

title: The gown and the korowai: Maori doctoral students and the spatial organization of academic knowledge

authors: Middleton, Sue

subjects: culture

abstract: This paper draws on 38 student interviews carried out in the course of the team research project Teaching and Learning in the Supervision of Māori Doctoral Students. Māori doctoral thesis work takes place in the intersections between the Māori (tribal) world of identifications and obligations, the organisational and epistemological configurations of academia, and the bureaucratic requirements of funding or employing bureaucracies. To explore how students accommodate cultural, academic and bureaucratic demands, we develop analytical tools combining three intellectual traditions: Māori educational theory, Bernstein’s sociology of the academy, and Lefebvre’s conceptual trilogy of perceived, conceived, and lived space.

text: The gown and the korowai: Maori doctoral students and the spatial organization of academic knowledge This paper draws on 38 student interviews carried out in the course of the team research project Teaching and Learning in the Supervision of Māori Doctoral Students. Māori doctoral thesis work takes place in the intersections between the Māori (tribal) world of identifications and obligations, the organisational and epistemological configurations of academia, and the bureaucratic requirements of funding or employing bureaucracies. To explore how students accommodate cultural, academic and bureaucratic demands, we develop analytical tools combining three intellectual traditions: Māori educational theory, Bernstein’s sociology of the academy, and Lefebvre’s conceptual trilogy of perceived, conceived, and lived space.

year: 2009