부산대학교 도서관

English: Yarrowia lipolytica is a non-conventional yeast which is considered to be suitable for production of industrially important proteins at commercial scale. The yeast is non pathogenic and is generally regarded as safe and as such can be used to produce biotechnological products for human consumption and applications. The genetic tools for the manipulation of the yeast have been developed over the years. This includes chemical and genetic based mutagenesis techniques to develop and isolate Y. lipolytica strains with enhanced properties in the production of endogenous and heterologous proteins. Protein expression systems comprised of a variety of inducible and constitutive promoters are available; secretion signals for extracellular production of proteins and a choice of replicative or integrative expression systems have also been developed. Auxotrophic and antibiotic based selectable markers are available some of which result in single or multiple copies of the integrative expression cassettes containing the target gene. The Y. lipolytica yeast can utilise a wide variety of carbon sources and its fermentation processes in bioreactors are well established. The property of Y. lipolytica as a prolific producer of the endogenous extracellular Lip2p was exploited for the development of an expression system for production of therapeutic peptides. To this end, the 38 amino acid long RANTES I peptide with therapeutic applications in the treatment of HIV/AIDS was selected as a model peptide for co-expression with Lip2p. The sequence encoding the peptide was cloned downstream of the complete LIP2 gene. The therapeutic peptide was separated from the LIP2 gene by the sequences encoding for 6X His and the DDDDK sequence recognized by the enterokinase proteolytic enzyme. The expression cassette was under the hp4d quasi constitutive promoter with the rDNA as a target for multi copy integration using the defective Ura3d4 integration cassette. The amount of RANTES I produced b
Afrikaans: Yarrowia lipolytica is ‘n onkonvensionele gis wat beskou word as geskik vir die produksie van proteïene van industriële belang op kommersiële skaal. Die gis is niepatogenies en word algeneem beskou as veilig (generally regarded as safe) en kan gevolglik gebruik word vir die produksie van biotegnologiese produkte vir menslike verbruik. Die genetiese gereedskap vir die manupilasie van gis is oor die jare ontwikkel. Dit sluit in chemies- en geneties-gebasseerde mutagenese tegnieke om Y. lipolytica rasse te ontwikkel en isoleer met verbeterde eienskappe vir produksie van endogene en heteroloë proteïene. Proteïenuitdrukkingsisteme, bestaande uit ‘n verskeidenheid induseerbare en konstitutiewe promotors, is beskikbaar; sekresieseine vir ekstrasellulêre produksie van proteïene asook ‘n keuse van replikatiewe of integrerende uitdrukkingsisteme is ook al ontwikkel. Ouksotrofiese en antibiotika gebasseerde selekteerbare merkers is beskikbaar, waarvan sommige lei tot integrasie van enkel of veelvuldige kopieë van die integrerende uitdrukkingskasette wat die teikengeen bevat. Die Y. lipolytica gis kan ‘n wye verskeidenheid koolstofbronne benut en het ‘n goed vasgestelde fermentasieproses in bioreaktors. Die eienskap van Y. lipolytica as ‘n effektiewe produseerder van endogeniese ekstrasellulêre Lip2p is gebruik vir die ontwikkeling van ‘n uitdrukkingsisteem vir die produksie van terapeutiese proteïene. Vir hierdie doel is die 38 aminosuur lange RANTES I peptied, met terapeutiese toepassings in die behandeling van HIV/VIGS, gekies as ‘n model peptied vir ko-uitdrukking met die Lip2p. Die volgorde wat kodeer vir die peptied is stroom-af van die volledige LIP2 geen gekloneer. Die terapeutiese peptied is geskei van die LIP2 geen deur die volgordes wat kodeer vir 6X His en die DDDK volgorde wat deur enterokinase ensiem herken word. Die uitdrukkingskaset is onder beheer van die hp4d quasi-konstitutiewe promotor met die rDNA as ‘n teiken vir veelvuldige integrasie deur gebr
CSIR and NRF