Biology subjects

Lutz, L.

Publications and source records attributed to Lutz, L..

4 recordsLinked to original sources

bioRxiv2026

Alcohol dehydrogenase-mediated methanol dissimilation increases carbon efficiency in synthetic autotrophic yeast

The efficient production of food and biochemicals using microorganisms that utilize single-carbon feedstocks presents a promising approach for advancing a circular bioeconomy. Komagataella phaffii (formerly Pichia pastoris) is a methylotrophic yeast already widely used in industry, making it an attractive host for such applications. Recently, K. phaffii was converted into an autotrophic strain capable of assimilating CO2 into both biomass and secreted organic acids, using energy derived from dissimilation of methanol to CO2. In these strains, methanol oxidation is catalysed by an alcohol oxidase (Aox2), which transfers electrons to oxygen without conserving reducing equivalents. To address this limitation, in this study we explored redirecting methanol dissimilation through the native alcohol dehydrogenase (Adh2), coupling methanol oxidation with NADH generation to improve carbon efficiency. By deleting AOX2 and overexpressing ADH2, we generated Adh2-based autotrophic strains that exhibited growth rates comparable to the parental strain (0.007 h-{superscript 1}), while reducing specific CO2 production by 53% and increasing biomass yield (YX/MeOH) by 59%. We further applied this strategy to convert previously developed autotrophic strains producing itaconic acid and lactic acid into Adh2-dependent strains. Optimizing ADH2 expression through multicopy integration resulted in strains with approximately two-fold higher molar carbon efficiency (Y(X+P)/CO2) while achieving elevated product titers--2.2-fold for itaconic acid and 3.8-fold for lactic acid--relative to the parental strains. Our findings demonstrate that alcohol dehydrogenase-mediated methanol dissimilation can significantly improve yield and productivity of autotrophic K. phaffii strains, with broad implications for sustainable bioproduction from one-carbon substrates.

biochemistry↗

bioRxiv2025

Parental exposure to wet and dry conditions shapes the viability and thermotolerance of eggs in Aedes aegypti

Aedes aegypti, a primary vector of dengue, Zika, and chikungunya, displays remarkable adaptability across ecological gradients. Central to this resilience is the egg stage, which must withstand fluctuating moisture and temperature conditions. Environmental transitions, particularly changes in moisture availability, significantly influence egg hatching success in mosquitoes. This study investigates how parental exposure to variable hydration conditions shapes key reproductive traits in Ae. aegypti. Using four environmental regimes, continuous wet, continuous dry, wet-to- dry, and dry-to-wet, we assessed egg output, hatching success, thermotolerance, and egg nutrient composition across three Ae. aegypti populations. Our results show that oviposition timing and egg production are significantly affected by the hydration environment experienced by the parental generation. While the wet, dry, and dry-to-wet groups exhibited a consistent oviposition peak beginning four days post-blood feeding, the wet-to-dry group showed delayed reproductive investment, with peak egg production occurring later. Egg output was highest under continuous wet conditions and significantly reduced in the dry and wet-to-dry treatments across all populations. Interestingly, the wet-to-dry group showed significantly higher egg-thermotolerance than any other group, and this pattern was consistent across all three populations under high- temperature stress conditions (41{degrees}C and 45{degrees}C). Nutritional composition showed an increased glycogen level in eggs when parents were exposed to wet conditions before blood feeding. By integrating physiological and ecological metrics such as hatching rates and thermal stress resilience, we demonstrate how parental environments shape subsequent egg performance, highlighting adaptive responses that enable Ae. aegypti persistence under increasing climate variability.

physiology↗

bioRxiv2025

Synthetic autotrophic yeast enables high itaconic acid production from CO2 via integrated pathway and process design

Single carbon (C1) substrates are gaining importance as future feedstocks for the production of bio-based chemicals. Carbon dioxide, a major greenhouse gas, offers a promising alternative to the traditional feedstocks to shift towards C1-based, sustainable processes. Here, we present a synthetic autotrophic Komagataella phaffii (Pichia pastoris) that is able to produce itaconic acid by the direct conversion of CO2, achieving final titers of approximately 12 g L-1 in bioreactor cultivations. We show that a combined approach that integrates balancing the flux between the Calvin-Benson-Bassham (CBB) cycle and itaconic acid metabolism with process design was essential to enhance the production. Our study demonstrates the potential of K. phaffii as a microbial platform using CO2 as the direct carbon source, aligning with the future goals of establishing sustainable bioprocesses.

synthetic biology↗

bioRxiv2025

Integration of strain and process optimization to increase autotrophic growth of engineered Komagataella phaffii

Synthetic autotrophs are a promising platform for sustainable bioproduction using CO2 as substrate. The methylotrophic yeast Komagataella phaffii has been engineered to use CO2 as the sole carbon source by integration of the Calvin-Benson-Bassham (CBB) cycle, based on its native methanol assimilating xylulose monophosphate pathway (XuMP) cycle. Initial growth rates were low, but could be doubled by adaptive laboratory evolution (ALE). Beneficial mutations led to a decrease of CBB cycle reactions, indicating further limitations. During this study, temperature was identified as one of the key process parameters to improve autotrophic growth. For this reason, a new round of adaptive laboratory evolution was performed at the identified optimal cultivation temperature of 25{degrees}C, resulting in isolates growing up to 50 % faster compared to the control strain. Whole genome resequencing followed by reverse engineering helped to identify first key mutations of the evolved strains. In addition, targeted engineering was performed by increasing the copy number of the key gene of the CBB cycle RuBisCO, which is the bottleneck of carbon fixation. Combining this with the optimal cultivation temperature boosted maximum specific growth rates of the autotrophic K. phaffii strain. In comparison to ALE, the targeted engineering still is lagging behind a bit. Starting from the initial condition, growth was boosted more than 2.5-fold in this study to a maximum of 0.025 h-1.

synthetic biology↗