Biology subjects

Lammers, P. J.

Publications and source records attributed to Lammers, P. J..

3 recordsLinked to original sources

bioRxiv2025

Elevated carbon dioxide stimulates highly efficient organic-carbon consumption and confectionary-waste valorization under mixotrophy in the unicellular alga Galdieria

Unicellular algae are appealing for nutritional and biotechnological utility but have wide variation across strains and can be challenging to produce. The thermo-acidophilic algal genus Galdieria use diverse organic-carbon sources for fermentative growth that can include waste-stream feedstocks and have complete amino-acid compositions for human nutrition. Here, we investigated Galdieria metabolic dynamics to catalog organic-carbon conversion to biomass. Tested strains had enhanced growth upon 3% CO2 supplementation, triggering efficient glucose uptake to reach [~]5 {+/-} 0.3 g dry biomass L-{superscript 1}. Stable-isotope analysis revealed that organic-carbon uptake dominates CO2 fixation in darkness under mixotrophy, with CO2 an apparent metabolic trigger. Galdieria sulphuraria 5587.1 can consume up to 8.3 g carbon L-1 day-1 from industrial confectionery waste, with C-phycocyanin reaching 3.8% of dry biomass and remaining thermostable at 72{degrees}C. This framework can optimize Galdieria-based bioprocesses for inexpensive waste conversion into high-value biomass and identifies CO2 as a trigger of organic-carbon assimilation, even in heterotrophic conditions. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=135 SRC="FIGDIR/small/655468v1_ufig1.gif" ALT="Figure 1"> View larger version (37K): org.highwire.dtl.DTLVardef@e2ef63org.highwire.dtl.DTLVardef@d28574org.highwire.dtl.DTLVardef@137be97org.highwire.dtl.DTLVardef@c1c0a5_HPS_FORMAT_FIGEXP M_FIG C_FIG

bioengineering↗

bioRxiv2024

A rapid CAT transformation protocol and nuclear transgene expression tools for metabolic engineering in Cyanidioschyzon merolae 10D

The eukaryotic red alga Cyanidioschyzon merolae 10D is an emerging algal host for synthetic biology and metabolic engineering. Its small nuclear genome (16.5 Mb; 4775 genes), low intron content (38), stable transgene expression, and capacity for homologous recombination into its nuclear genome make it ideal for genetic and metabolic engineering endeavors. Here, we present an optimized transformation and selection protocol, which yields single chloramphenicol-resistant transformants in under two weeks. Transformation dynamics and a synthetic modular plasmid toolkit are reported, including several new fluorescent reporters. Techniques for fluorescence reporter imaging and analysis at different scales are presented to facilitate high-throughput screening of C. merolae transformants. We use this plasmid toolkit to overexpress the Ipomoea batatas isoprene synthase and demonstrate the dynamics of engineered volatile isoprene production during different light regimes using multi-port headspace analysis coupled to parallel photobioreactors. This work seeks to promote C. merolae as an algal system for metabolic engineering and future sustainable biotechnological production.

bioengineering↗

bioRxiv2023

Engineered ketocarotenoid biosynthesis in the polyextremophilic red microalga Cyanidioschyzon merolae 10D

The polyextremophilic Cyanidiales are eukaryotic red microalgae with promising biotechnological properties arising from their low pH and elevated temperature requirements which can minimize culture contamination at scale. Cyanidioschyzon merolae 10D is a cell wall deficient species with a fully sequenced genome that is amenable to nuclear transgene integration by targeted homologous recombination. C. merolae maintains a minimal carotenoid profile and here, we sought to determine its capacity for ketocarotenoid accumulation mediated by heterologous expression of a green algal {beta}-carotene ketolase (BKT) and hydroxylase (CHYB). To achieve this, a synthetic transgene expression cassette system was built to integrate and express Chlamydomonas reinhardtii (Cr) sourced enzymes by fusing native C. merolae transcription, translation and chloroplast targeting signals to codon-optimized coding sequences. Chloramphenicol resistance was used to select for the integration of synthetic linear DNAs into a neutral site within the host genome. CrBKT expression caused accumulation of canthaxanthin and adonirubin as major carotenoids while co-expression of CrBKT with CrCHYB generated astaxanthin as the major carotenoid in C. merolae. Unlike green algae and plants, ketocarotenoid accumulation in C. merolae did not reduce total carotenoid contents, but chlorophyll a reduction was observed. Light intensity affected global ratios of all pigments but not individual pigment compositions and phycocyanin contents were not markedly different between parental strain and transformants. Continuous illumination was found to encourage biomass accumulation and all strains could be cultivated in simulated summer conditions from two different extreme desert environments. Our findings present the first example of carotenoid metabolic engineering in a red eukaryotic microalga and open the possibility for use of C. merolae 10D for simultaneous production of phycocyanin and ketocarotenoid pigments. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=71 SRC="FIGDIR/small/530181v2_ufig1.gif" ALT="Figure 1"> View larger version (27K): org.highwire.dtl.DTLVardef@dd010dorg.highwire.dtl.DTLVardef@1700e5corg.highwire.dtl.DTLVardef@1bee8eaorg.highwire.dtl.DTLVardef@ad67da_HPS_FORMAT_FIGEXP M_FIG C_FIG

bioengineering↗