A5088267094- Pharmacogenetics and Drug Metabolism
- Microbial Metabolic Engineering and Bioproduction
- Biochemical and biochemical processes
- Metal-Catalyzed Oxygenation Mechanisms
- Plant biochemistry and biosynthesis
- Fungal and yeast genetics research
- Steroid Chemistry and Biochemistry
- Enzyme Catalysis and Immobilization
- Enzyme-mediated dye degradation
- Plant Parasitism and Resistance
- Plant Physiology and Cultivation Studies
- Inflammatory mediators and NSAID effects
- Viral Infectious Diseases and Gene Expression in Insects
- Chemical Reactions and Isotopes
- Edible Oils Quality and Analysis
- Plant-Microbe Interactions and Immunity
- Nitric Oxide and Endothelin Effects
- Piperaceae Chemical and Biological Studies
- CRISPR and Genetic Engineering
- Bacterial Genetics and Biotechnology
- Genetically Modified Organisms Research
- Biotin and Related Studies
- Microbial Natural Products and Biosynthesis
- Oxidative Organic Chemistry Reactions
Austrian Centre of Industrial Biotechnology (Austria)
2019-2023
Graz University of Technology
2021
Cell Biotech (South Korea)
2018-2021
Institute of Molecular Biotechnology
2021
Since 2004, unspecific peroxygenases, in short UPOs (EC. 1.11.2.1), have been explored. are closing a gap between P450 monooxygenases and chloroperoxidases. These enzymes highly active biocatalysts for the selective oxyfunctionalisation of C–H, C=C C-C bonds. secreted fungal proteins Komagataella phaffii (Pichia pastoris) is an ideal host high throughput screening approaches UPO production. Heterologous overexpression 26 new by K. was performed deep well plate cultivation shake flask up to...
Unspecific peroxygenases (UPOs) are glycosylated fungal enzymes that can selectively oxidize C–H bonds. UPOs employ hydrogen peroxide as the oxygen donor and reductant. With such an easy-to-handle cosubstrate without need for a reducing agent, emerging convenient oxidative biocatalysts. Here, unspecific peroxygenase from Hypoxylon sp. EC38 (HspUPO) was identified in activity-based screen of six putative were heterologously expressed Pichia pastoris. The enzyme found to tolerate selected...
Green leaf volatiles (GLVs) cover a group of mainly C6-and C9-aldehydes, -alcohols and -esters. Their name refers to their characteristic herbal fruity scent, which is similar that freshly cut grass or vegetables. Lipoxygenases (LOXs) catalyze the peroxidation unsaturated fatty acids. The resulting hydroperoxy acids are then cleaved into aldehydes oxo by acid hydroperoxide lyases (HPLs). Herein, we equipped yeast Komagataella phaffii with recombinant genes coding for LOX HPL, serve as...
The industrially important non-conventional yeast Komagataella phaffii suffers from low rates of homologous recombination, making site specific genetic engineering tedious. Therefore, genome editing using CRISPR/Cas represents a simple and efficient alternative. To characterize on- off-target mutations caused by CRISPR/Cas9 followed non-homologous end joining repair, we chose diverse set targets conducted whole sequencing on 146 engineered single colonies. We compared the outcomes target...
Abstract Background Currently, the numerous and versatile applications in pharmaceutical chemical industry make recombinant production of cytochrome P450 enzymes (CYPs) great biotechnological interest. Accelerating drug development process by simple, quick scalable access human metabolites is key for efficient targeted response to new sometimes unexpected medical challenges needs. However, due its biochemical complexity, CYP (hCYP) their application preparative biotransformations was still...
Abstract Class VIII self-sufficient CYP from Aspergillus fumigatus (CYP505X) and a variant with five mutations were heterologously expressed in Pichia pastoris. Ibuprofen – widely used non-steroidal anti-inflammatory drug (NSAID) was oxidized most efficiently whole cell mediated biotransformation. The two major metabolites isolated by preparative HPLC their structure elucidated NMR. Specifically, ibuprofen hydroxylated at the tertiary carbon atom benzylic position on lipophilic side chain.
Abstract Among nature's arsenal of oxidative enzymes, cytochrome P450s (CYPs) catalyze the most challenging reactions, hydroxylations non‐activated C−H bonds. Human CYPs are studied in drug development due to their physiological role at forefront metabolic detoxification, but handling makes them unsuitable for application. have a great potential biocatalysis, often lack appropriate features such as high and soluble expression, self‐sufficient internal electron transport, stability, an...
Pseudomonas aeruginosa lipoxygenase (PaLOX) catalyzes the peroxidation of unsaturated fatty acids. Not only linoleic acid, but also linolenic acid and oleic are oxidized. The natural host secretes PaLOX into periplasmic space. Herein, aim is to secrete culture supernatant Pichia pastoris. Since protein background in typically rather low, this strategy allows for almost pure production applicable valorization renewable acids, example green leaf volatiles. Using CAT1 promoter system...