Purification, crystallization and cloning of tributyrin esterase from Lactococcus : a thesis presented in partial filfillment of the requirements for the degree of Master of Technology in Biotechnology in the Institute of Molecular Biosciences at Massey University, New Zealand

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dc.contributor.authorZheng, Jiong
dc.date.accessioned2018-06-25T21:57:48Z
dc.date.available2018-06-25T21:57:48Z
dc.date.issued1998
dc.description.abstractTributyrin esterase is an enzyme that has been isolated and purified from lactococcal starter strain by research staff at the New Zealand Dairy Research Institute. It has been shown to play an important role in production and control of flavour development during cheese ripening, but little is known about its biochemical characteristics. New studies on tributyrin esterase have been initiated, with the aim of carrying out a three dimensional structure determination to completely understand the molecular basis and the nature of its in vivo activity. This thesis is divided into three main parts. In the first part, the purification of tributyrin esterase from a genetically modified strain Lc. lactis subsp. cremoris B1079 is described. The procedure investigated for optimization of the protocol and a partial study of factors affecting tributyrin esterase activity are described. In the second part, crystallization trials for tributyrin esterase are described. Several crystals were obtained, with the best ordered crystals being grown from 2.6M ammonium sulfate, these have been shown to diffract to 3.0 Å, and belong to the space group C222 with cell dimensions a=76Å, b=178Å c=179Å. In the third part, the lipase gene was ligated into 4.75kbp expression vector proEX, which contains a his-tag sequence upstream of the multiple cloning site. The ligation reaction mixture was transformed into competent E.coli DH5α cells. This should allow the expression of tributyrin esterase in E.coli and eventually provide a great yield of protein and make purification easier.en_US
dc.identifier.urihttp://hdl.handle.net/10179/13549
dc.language.isoenen_US
dc.publisherMassey Universityen_US
dc.rightsThe Authoren_US
dc.subjectLactococcus lactisen_US
dc.subjectEsterasesen_US
dc.subjectEnzymes -- Biotechnologyen_US
dc.titlePurification, crystallization and cloning of tributyrin esterase from Lactococcus : a thesis presented in partial filfillment of the requirements for the degree of Master of Technology in Biotechnology in the Institute of Molecular Biosciences at Massey University, New Zealanden_US
dc.typeThesisen_US
massey.contributor.authorZheng, Jiong
thesis.degree.disciplineBiotechnologyen_US
thesis.degree.grantorMassey Universityen_US
thesis.degree.levelMastersen_US
thesis.degree.nameMaster of Technology (M. Tech.)en_US
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