Employment of alkane 1-monooxygenase in efficient whole-cell production of tulipalin A

master's thesis

• Title (croatian): Primjena alkan-1-monooksigenaze u učinkovitoj cjelostaničnoj proizvodnji tulipalina A
• Author: Marina Grgić
• Mentor: Anita Slavica (mentor)
• Committee member: Renata Teparic (predsjednik povjerenstva)
• Committee member: Blaženka Kos (član povjerenstva)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Biochemical Engineering); (Laboratory for Biochemical Engineering, Industrial Microbiology and Malting and Brewing Technology); Zagreb
• Defense date and country: 2023-07-18, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Alkane 1-monooxygenases (Alk) were investigated in order to optimize the conversion of isoprenyl acetate (MBAc) to 3-(hydroxymethyl)but-3-en-1-yl acetate (HMBAc), as a part of the artificial synthetic pathway towards tulipalin A. Hydroxylation of MBAc to HMBAc and n-octane to 1-octanol and 1-octanoic acid, by employed Alk enzymes expressed in E. coli BL21(DE3) cells was more efficient at 25 °C than at 30 or 20 °C. Co-expression of a transporter of the hydrophobic substrate MBAc - AlkL with Alk from Marinobacter hydrocarbonoclasticus SP17 (MhAlkMO) has improved the uptake and conversion of MBAc by 1.4-fold. The highest specific activity toward the substrate was estimated for the mutant MhAlkMO-I238V (3.61 U gcdw⁻¹ ), which is a 2.8-fold improvement in comparison to the originally tested Alk from Pseudomonas putida GPo1 (AlkB).

Abstract (croatian)

Istražena je primjena alkan 1-monooksigenaza (Alk) u cilju optimizacije konverzije izoprenil acetata (MBAc) do 3-(hidroksimetil)but-3-en-1-il acetata (HMBAc), koja je dio kreiranog puta sinteze tulipalina A. Hidroksilacija MBAc do HMBAc kao i n-oktana do 1-oktanola i 1-oktanske kiseline odabranim Alk enzimima, koji su eksprimirani u stanicama E. coli BL21(DE3), bila je učinkovitija pri 25 °C nego pri 30 ili 20 °C. Ko-ekspresija transportera hidrofobnog susptrata MBAc - AlkL sa Alk iz Marinobacter hydrocarbonoclasticus SP17 (MhAlkMO) poboljšala je transport i konverziju ovoga supstrata za 1.4 puta. Najveća specifična aktivnost prema MBAc procijenjena je za mutant MhAlkMO-I238V (3.61 U gcdw⁻¹), što je poboljšanje specifične aktivnosti od 2.8-puta u usporedbi s prvobitno testiranom Alk iz Pseudomonas putida GPo1 (AlkB).

• Keywords: tulipalin A; alkane 1-monooxygenase; transporter of the hydrophobic substrate; whole-cell biotransformation
• Keywords (croatian): tulipalin A; alkan 1-monooksigenaza; transporter hidrofobnog supstrata; cjelostanična biotransformacija
• Language: english
• URN:NBN: urn:nbn:hr:159:471414
• Study programme: Title: Molecular Biotechnology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra inženjer/inženjerka molekularne biotehnologije (magistar/magistra inženjer/inženjerka molekularne biotehnologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2024-06-13 08:28:21