Heterologno izražanje genov terpenoidne biosintezne poti v izbranih sevih Streptomyces sp.

Uranič, Eva (author), Petković, Hrvoje (mentor), Paš, Maja (comentor)

Bakterije rodu Streptomyces so znane predvsem po produkciji sekundarnih metabolitov. V zadnjih letih industrija kaže veliko zanimanja tudi za produkcijo različnih metabolitov v industrijskih heterolognih gostiteljih. Cilj magistrske naloge je bil heterologno izraziti gene za biosintezo izopentenil difosfata (IPP) in gene za pretvorbo IPP v β-karoten v izbranih sevih Streptomyces sp. Konstruirali smo plazmide z genskim zapisom za mevalonatno pot (MEV) in plazmide z geni za biosintezo β-karotena. Genske zapise za MEV pot smo pridobili iz dveh bakterijskih vrst družine Streptomycetaceae in sicer Streptomyces cinnamonensis (MEVC) in Kitasatospora griseola (MEVK). Geni za biosintezo β-karotena, ki smo jih uporabili, so iz bakterije Erwinia uredova. Za konstrukcijo plazmidov smo uporabili več vektorjev, in sicer integrativne, ki vsebujejo virusno integrazo phiBT in phiC31 in E. coli-Streptomyces replikativni vektor pVF. Konstruirali smo plazmide z geni za pot MEV in jih transformirali v bakterijo S. rimosus white. V obsegu te naloge smo poskusili hkrati transformirati tudi plazmide z geni za biosintezo β-karotena, vendar nam to ni uspelo.

• Language: Slovenian
• Keywords: Streptomyces rimosus, heterologno izražanje, biosinteza, β-karoten, mevalonatna pot, pot deoksiksiluloza-5-fosfata, izopentenil difosfat, plazmidi
• Work type: Master's thesis/paper
• Typology: 2.09 - Master's Thesis
• Organization: BF - Biotechnical Faculty
• Publisher: [E. Uranič]
• Year: 2018
• PID: 20.500.12556/RUL-101965
• UDC: 602.3+602.6:579.873.7:577.1
• COBISS.SI-ID: 4930680
• Publication date in RUL: 15.07.2018

Secondary language

• Language: English
• Title: Heterologus expression of genes for terpenoide biosynthetic pathways in selected Streptomyces sp. strains

Abstract

Bacteria of genus Streptomyces are well known secondary metabolites producers. Streptomyces species are also attractive host for heterologous production of various metabolites in industrial level. The aim of our thesis was to express genes for biosynthesis of isopentenyl diphosphate (IPP) and genes for conversion of IPP in βcatotene in Streptomyces sp. Plasmids with mevalonate (MEV) and with β-carotene pathway were constructed. Genes for MEV pathway were extracted from two Streptomyces species; S. cinnamonensis (MEVC) and K. griseola (MEVK). Genes for β- carotene biosynthesis were used from bacteria Erwinia uredova. For plasmid construction we used integrative vectors with phiBT and phiC31 virus integrase and E. coli-Streptomyces replicative vector pVF. We engineered plasmids with MEV pathway and transformed them into S. rimosus successfully. We also try to transform plasmids with β-carotene pathway but were unsuccesful.

• Keywords: Streptomyces rimosus, heterologous expression, biosynthesis, β-caroten, mevalonat pathway, deoxysilulose-5-phosphate pathway, isopentenyl diphosphate, plasmid