Searched for: subject:"Mutagenesis%2C%5C+Site%5C-Directed"
(1 - 17 of 17)
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Ruijssenaars, H.J. (author), Sperling, E.M.G.M. (author), Wiegerinck, P.H.G. (author), Brands, F.T.L. (author), Wery, J. (author), de Bont, J.A.M. (author), TNO Kwaliteit van Leven (author)
A steroid 15β-hydroxylating whole-cell solvent tolerant biocatalyst was constructed by expressing the Bacillus megaterium steroid hydroxylase CYP106A2 in the solvent tolerant Pseudomonas putida S12. Testosterone hydroxylation was improved by a factor 16 by co-expressing Fer, a putative Fe-S protein from Bacillus subtilis. In addition, the...
article 2007
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Michielse, C.B. (author), Hooykaas, P.J.J. (author), van den Hondel, C.A.M.J.J. (author), Ram, A.F.J. (author), TNO Kwaliteit van Leven (author)
In the era of functional genomics, the need for tools to perform large-scale targeted and random mutagenesis is increasing. A potential tool is Agrobacterium-mediated fungal transformation. A. tumefaciens is able to transfer a part of its DNA (transferred DNA; T-DNA) to a wide variety of fungi and the number of fungi that can be transformed by...
article 2005
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TNO Kwaliteit van Leven (author), Kypreos, K.E. (author), van Dijk, K.W. (author), Havekes, L.M. (author), Zannis, V.I. (author)
To identify the residues in the carboxyl-terminal region 260-299 of human apolipoprotein E (apoE) that contribute to hypertriglyceridemia, two sets of conserved, hydrophobic amino acids between residues 261 and 283 were mutated to alanines, and recombinant adenoviruses expressing these apoE mutants were generated. Adenovirus-mediated gene...
article 2005
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Kralj, S. (author), van Geel-Schutten, G.H. (author), van der Maarel, M.J.E.C. (author), Dijkhuizen, L. (author), TNO Voeding Centraal Instituut voor Voedingsonderzoek TNO (author)
Lactobacillus reuteri strain 121 uses sucrose for synthesis of a unique, soluble glucan ('reuteran') with mainly α-(1→4) glucosidic linkages. The gene (gtfA) encoding this glucansucrase enzyme had previously been characterized. Here, a detailed biochemical and molecular analysis of the GTFA enzyme is presented. This is believed to be the first...
article 2004
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TNO Voeding Centraal Instituut voor Voedingsonderzoek TNO (author), Ozimek, L.K. (author), van Hijum, S.A.F.T. (author), van Koningsveld, G.A. (author), van der Maarel, M.J.E.C. (author), van Geel-Schutten, G.H. (author), Dijkhuizen, L. (author)
Bacterial fructosyltransferases (FTFs) are retaining-type glycosidases that belong to family 68 of glycoside hydrolases. Recently, the high-resolution 3D structure of the Bacillus subtilis levansucrase has been solved [Meng, G. and Futterer, K., Nat. Struct. Biol. 10 (2003) 935-941]. Based on this structure, the catalytic nucleophile, general...
article 2004
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Arand, M. (author), Hallberg, B.M. (author), Zou, J. (author), Bergfors, T. (author), Oesch, F. (author), van der Werf, M.J. (author), de Bont, J.A.M. (author), Jones, T.A. (author), Mowbray, S.L. (author)
Epoxide hydrolases are essential for the processing of epoxide-containing compounds in detoxification or metabolism. The classic epoxide hydrolases have an α/β hydrolase fold and act via a two-step reaction mechanism including an enzyme-substrate intermediate. We report here the structure of the limonene-1,2-epoxide hydrolase from Rhodococcus...
article 2003
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Smit, E. (author), Jager, D. (author), Martinez, B. (author), Tielen, F.J. (author), Pouwels, P.H. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)
The structure of the crystallisation domain, SAN, of the S A-protein of Lactobacillus acidophilus ATCC 4356 was analysed by insertion and deletion mutagenesis, and by proteolytic treatment. Mutant S A-protein synthesised in Escherichia coli with 7-13 amino acid insertions near the N terminus or within regions of sequence variation in SAN (amino...
article 2002
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Gaubius Instituut TNO (author), Gervois, P. (author), Vu-Dac, N. (author), Kleemann, R. (author), Kockx, M. (author), Dubois, G. (author), Laine, B. (author), Kosykh, V. (author), Fruchart, J.C. (author), Kooistra, T. (author), Staels, B. (author)
Fibrinogen is a coagulation factor and an acute phase reactant up-regulated by inflammatory cytokines, such as interleukin 6 (IL-6). Elevated plasma fibrinogen levels are associated with coronary heart diseases. Fibrates are clinically used hypolipidemic drugs that act via the nuclear receptor peroxisome proliferator-activated receptor alpha ...
article 2001
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Ordentlich, A. (author), Barak, D. (author), Kronman, C. (author), Benschop, H.P. (author), de Jong, L.P.A. (author), Ariel, N. (author), Barak, R. (author), Segall, Y. (author), Velan, B. (author), Shafferman, A. (author)
The stereoselectivity of the phosphonylation reaction and the effects of adduct configuration on the aging process were examined for human acetylcholinesterase (HuAChE) and its selected active center mutants, using the four stereomers of 1,2,2-trimethylpropyl methylphosphonofluoridate (soman). The reactivity of wild type HuAChE toward the P(S)...
article 1999
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van Gemeren, I.A. (author), Beijersbergen, A. (author), van den Hondel, C.A.M.J.J. (author), Verrips, C.T. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)
Several cutinase variants derived by molecular modelling and site- directed mutagenesis of a cutinase gene from Fusarium solani pisi are poorly secreted by Saccharomyces cerevisiae. The majority of these variants are successfully produced by the filamentous fungus Aspergillus awamori. However, the L51S and T179Y mutations caused reductions in...
article 1998
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Quax, P.H.A. (author), Grimbergen, J.M. (author), Lansink, M. (author), Bakker, A.H.F. (author), Blatter, M.C. (author), Belin, D. (author), van Hinsbergh, V.W.M. (author), Verheijen, J.H. (author), Gaubius Instituut TNO (author)
Urokinase-type plasminogen activator (UPA), particularly when bound to its receptor (UPAR), is thought to play a major role in local proteolytic processes, thus facilitating cell migration as may occur during angiogenesis, neointima and atherosclerotic plaque formation, and tumor cell invasion. To facilitate understanding of the need and...
article 1998
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Gaubius Instituut TNO (author), Vu-Dac, N. (author), Gervois, P. (author), Torra, I.P. (author), Fruchart, J.C. (author), Kosykh, V. (author), Kooistra, T. (author), Princen, H.M.G. (author), Dallongeville, J. (author), Staels, B. (author)
Hypertriglyceridemia is a metabolic complication of retinoid therapy. In this study, we analyzed whether retinoids increase the expression of apo C- III, an antagonist of plasma triglyceride catabolism. In men, isotretinoin treatment (80 mg/d; 5 d) resulted in elevated plasma apo C-III, but not apo E concentrations. In human hepatoma HepG2 cells...
article 1998
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Gaubius Instituut TNO (author), Verheijen, J.H. (author), Nieuwenbroek, N.M.E. (author), Beekman, B. (author), Hanemaaijer, R. (author), Verspaget, H.W. (author), Ronday, K.H. (author), Bakker, A.H.F. (author)
We describe a new principle for assessment of the activity of proteolytic enzymes of all classes and show the application of this principle for the quantitative assay of bacterial collagenase and human matrix metalloproteinases (MMPs). Central to this new principle is the presence of a proenzyme that can be activated into an active enzyme by a...
article 1997
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Jespersen, S. (author), Ploemen, J.H.T.M. (author), Bladeren, P.J. (author), Niessen, W.M.A. (author), Tjaden, U.R. (author), van der Greef, J. (author), Instituut CIVO-Toxicologie en Voeding TNO (author)
A mass spectrometric method providing qualitative site-specific information regarding covalent modification of proteins is described. The method involves comparison of unmodified and modified proteins by matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) peptide mapping in combination with site-specific mutagenesis of...
article 1996
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Gouka, R.J. (author), Hessing, J.G.M. (author), Stam, H. (author), Musters, W. (author), van den Hondel, C.A.M.J.J. (author), Centraal Instituut voor Voedingsonderzoek TNO (author)
A homologous gene transfer system for Aspergillus awamori for site-specific integration is described, based on two components. First, a defined A. awarnori pyrG mutant strain constructed by a selection strategy for gene-replacement in fungi. Second, a vector with a homologous pyrG selection marker containing a defined mutation at a site...
article 1995
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van Eijsden, R.R. (author), Hoedemaeker, F.J. (author), Díaz, C.L. (author), Lugtenberg, B.J.J. (author), de Sylvia Pater, B. (author), Kijne, J.W. (author)
As part of a strategy to determine the precise role of pea (Pisum sativum) lectin, Psl, in nodulation of pea by Rhizobium leguminosarum, mutations were introduced into the genetic determinant for pea lectin by site-directed mutagenesis using PCR. Introduction of a specific mutation, N125D, into a central area of the sugar-binding site resulted...
article 1992
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Weening-Verhoeff, E.J.D. (author), Quax, P.H.A. (author), van Leeuwen, R.T.J. (author), Rehberg, E.F. (author), Marotti, K.R. (author), Verheijen, J.H. (author), Gaubius Instituut TNO (author)
article 1990
Searched for: subject:"Mutagenesis%2C%5C+Site%5C-Directed"
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