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: Exploring Timorese family funds of knowledge and their use in education

authors: Soares Pinto, Zulmira Da Cruz

subjects: Timorese families

abstract: There are many factors that contribute to children’s development, including their social and cultural background. Families play a significant role in children’s development and the funds of knowledge that reside within a family and the wider community can be utilised in a preschool setting to make strong connections for children between their home and the preschool. Research suggests that including families’ funds of knowledge in the preschool curriculum can enhance a child’s engagement in the learning environment and lead to positive educational outcomes. Drawing on Moll and Amanti’s concept of funds of knowledge and using a qualitative methodology, this thesis examines notions of funds of knowledge in preschool settings in Timor Leste. Through the exploration of what funds of knowledge are held by Timorese families and how these may be used in a preschool classroom by teachers, the thesis argues that there is a need for further understanding and acceptance by teachers of the importance of recognising and including families’ funds of knowledge in the preschool classroom. The findings indicate that traditional attitudes about teachers being the authority in learning tend to prevail and this limits parents’ ability to contribute knowledge and assistance to the preschool classroom. There is clear evidence of family and communities’ holding a range of funds of knowledge that could be beneficial for children’s learning if included in classroom practices by teachers. The thesis calls for extensive professional opportunities for teachers to develop understanding about funds of knowledge and how they might make preschools more welcoming for parents.

text: Exploring Timorese family funds of knowledge and their use in education There are many factors that contribute to children’s development, including their social and cultural background. Families play a significant role in children’s development and the funds of knowledge that reside within a family and the wider community can be utilised in a preschool setting to make strong connections for children between their home and the preschool. Research suggests that including families’ funds of knowledge in the preschool curriculum can enhance a child’s engagement in the learning environment and lead to positive educational outcomes. Drawing on Moll and Amanti’s concept of funds of knowledge and using a qualitative methodology, this thesis examines notions of funds of knowledge in preschool settings in Timor Leste. Through the exploration of what funds of knowledge are held by Timorese families and how these may be used in a preschool classroom by teachers, the thesis argues that there is a need for further understanding and acceptance by teachers of the importance of recognising and including families’ funds of knowledge in the preschool classroom. The findings indicate that traditional attitudes about teachers being the authority in learning tend to prevail and this limits parents’ ability to contribute knowledge and assistance to the preschool classroom. There is clear evidence of family and communities’ holding a range of funds of knowledge that could be beneficial for children’s learning if included in classroom practices by teachers. The thesis calls for extensive professional opportunities for teachers to develop understanding about funds of knowledge and how they might make preschools more welcoming for parents.

year: 2018

title: Tāwharautia Aotearoa

authors: Maxwell, Te Kahautu

abstract: Tāwharautia Aotearoa was composed in opposition to the National Governments attempt to remove Section 9 of the Principles of the Treaty of Waitangi which would give the Government power to sell State assets. Performed by Ōpōtiki Mai Tawhiti at the Te Matatini World Championship Kapa Haa Competition 2013.

text: Tāwharautia Aotearoa Tāwharautia Aotearoa was composed in opposition to the National Governments attempt to remove Section 9 of the Principles of the Treaty of Waitangi which would give the Government power to sell State assets. Performed by Ōpōtiki Mai Tawhiti at the Te Matatini World Championship Kapa Haa Competition 2013.

year: 2013