Search bioRxiv⌕ Search

Biology subjects

Ledesma-Amaro, R.

Publications and source records attributed to Ledesma-Amaro, R..

16 recordsLinked to original sources

Adaptive laboratory evolution of a yeast co-culture chassis for modular bioproduction

Synthetic microbial consortia have the potential to bring novel architectures to biotechnological processes. They enable more flexible and efficient bioprocesses by reducing metabolic burden and supporting division of labour. Obligately mutualistic cross-feeding has been used to stabilise the population composition of artificially assembled consortia. However, many current strategies rely on the cross-feeding of metabolites with limited exchange rates, and this imposes a growth burden on the consortium members. Here, we used adaptive laboratory evolution (ALE) to address this growth bottleneck and create an optimised co-culture chassis to host bioproduction functions. Transcriptome analysis revealed how ALE alleviated stress responses associated with nutritional restrictions in the non-evolved cross-feeding system. Using a case-study split bioproduction pathway, we demonstrated that improvements in the chassis growth performance resulted in production improvements, surpassing the performance of a monoculture implementation by 1.5-fold. Our results show the potential of ALE to optimise the cross-feeding layer of yeast co-cultures, and how this enables the efficient implementation of a bioproduction process.

synthetic biology↗

Synthetic Yarrowia lipolytica consortium for efficient conversion of lignocellulosic oligosaccharides into lipids

Lignocellulosic biomass (LCB) is an abundant and renewable feedstock for the sustainable production of bioproducts; however, its industrial exploitation is limited by its complex composition and by the lack of microbial platforms capable of simultaneously degrading and assimilating cellulose- and hemicellulose-derived oligosaccharides. Yarrowia lipolytica lacks the native enzymatic machinery required for this process. In this study, we engineered a multifunctional strain (YBXT-XR-BGL3) able to secrete fungal {beta}-glucosidase (BGL3 or BGL1) and {beta}-xylosidase (BxTw1) from Talaromyces amestolkiae, enabling the hydrolysis of cellobiose and xylooligosaccharides, respectively. In addition, a xylose reductase pathway was introduced to confer xylose assimilation. Because the construction of a single multifunctional strain may impose a significant metabolic burden and reduce fitness, we benchmarked this strain against a division-of-labor strategy. To this end, we also developed a cellobiose-specialized strain (YBGL3 or YBGL1) and a xylooligosaccharide-specialized strain (YBXT-XR). Functional characterization revealed efficient saccharification of cello- and xylooligosaccharides under acidic conditions, with BGL3 outperforming BGL1 in glucose release and BxTw1 exhibiting broad pH tolerance. Under nitrogen-limited conditions, this enabled lipid accumulation of up to 20% from cellobiose in YBGL3 and YBXT-XR-BGL3, and up to 15% from xylooligosaccharides in YBXT and YBXT-XR-BGL3. In co-culture experiments using a mixed substrate (glucose, cellobiose, and xylooligosaccharides), both the multifunctional strain and the consortium produced up to 0.67 g L{square}{superscript 1} of lipids. However, the division-of-labor approach led to higher lipid accumulation (34% versus 26.3% in the monoculture), driven by a rapid population shift: following cellobiose depletion (after 72 h), YBXT-XR became predominant and utilized the remaining xylooligosaccharides almost exclusively for lipid synthesis. Overall, this study provides the first demonstration of a Y. lipolytica system capable of simultaneously utilizing cellulose- and hemicellulose-derived oligosaccharides. Moreover, benchmarking a division-of-labor consortium against a multifunctional monoculture highlights a robust strategy to enhance lipid biosynthesis and improve process resilience for LCB valorization.

synthetic biology↗

Engineering signal peptide-mediated recombinant protein secretion in Komagataeibacter rhaeticus for biological Engineered Living Material applications

BackgroundBacterial nanocellulose (BC), produced by Komagataeibacter species, is an ideal scaffold for biological Engineered Living Materials (bioELMs) research. Current BC functionalization strategies often rely on secondary microbial hosts or post-production enzyme immobilization, limiting the scalability and modularity required for programmable bioELMs. Establishing a single-chassis system capable of simultaneous biopolymer synthesis and in situ functionalization remains a primary objective in bioELM research. This study addresses the need by benchmarking signal peptide-mediated protein translocation in K. rhaeticus iGEM, a model bacterium for BC-based bioELMs, enabling a synthetic biology framework for single-chassis based biomaterial functionalization. ResultsGenome-wide analysis confirmed the presence of a complete Sec translocation machinery in K. rhaeticus. Through liquid chromatography-tandem mass spectrometry and SignalP 5.0 prediction, native signal peptides were identified and evaluated alongside previously characterized heterologous signal peptides using {beta}-lactamase and mScarlet as cargo proteins. Protein translocation was found to depend on signal peptide identity, cargo type, and expression mode. Fluorescence imaging revealed cytoplasmic, polar, and peripheral localization patterns, confirming functional engagement with the native translocation machinery. A key limitation identified was the retention of recombinant proteins within the periplasm, restricting extracellular availability. Despite this, signal peptide-mediated translocation enabled the incorporation of enzymatic activity into BC during biosynthesis. A post-growth osmotic shock-release strategy increased measurable enzymatic activity by 30%, demonstrating a practical route to overcome this physiological bottleneck while maintaining the biomaterial production capacity. ConclusionsThis study benchmarks signal peptide-dependent protein translocation in K. rhaeticus and identifies periplasmic retention as a key constraint for extracellular protein release. By linking protein translocation to in situ BC functionalization, this work establishes a synthetic biology framework that supports the development of K. rhaeticus as a single-chassis platform for the scalable production of functionalized bioELMs.

bioengineering↗

PromoterAtlas: decoding regulatory sequences across Gammaproteobacteria using a transformer model

Recent advances in deep learning, particularly transformer architectures, have improved computational approaches for biological sequence analysis. Despite these advances, computational models for bacterial promoter prediction have remained limited by small datasets, species-specific training, and binary classification approaches rather than comprehensive annotation frameworks. We present PromoterAtlas, a 1.8M parameter transformer model trained on 9M regulatory sequences from 3,371 gammaproteobacterial species. The model demonstrates recognition of various regulatory elements across different species, including ribosomal binding sites, various types of bacterial promoters, transcription factor binding sites, and terminators. Using this model, we developed a whole-genome promoter annotation tool for Gammaproteobacteria, with various levels of validation that support the predictions of promoters associated with different sigma ({sigma}) factors. Furthermore, we show that the model embeddings encode cross-species evolutionary relationships, clustering promoters by {sigma} factor identity rather than species-specific sequence features. Finally, we show that model embeddings encode regulatory sequence information that enables effective prediction of transcription and translation levels. PromoterAtlas can contribute to our understanding of and ability to engineer bacterial regulatory sequences with potential applications in bacterial biology, synthetic biology, and comparative genomics.

bioinformatics↗

Investigating The Potential of Division of Labour in Synthetic Bacterial Communities for the Production of Violacein

With advancements in synthetic biology and metabolic engineering, microorganisms can now be engineered to perform increasingly complex functions, which may be limited by the resources available in individual cells. Division of labour in synthetic microbial communities offers a promising approach to enhance metabolic efficiency and resilience in bioproduction. By distributing complex metabolic pathways across multiple subpopulations, the resource competition and metabolic burden imposed on an individual cell is reduced, potentially enabling more efficient production of target compounds. Violacein is a high-value pigment with anti-tumour properties that exemplifies such a challenge due to its complex bioproduction pathway, imposing a significant metabolic burden on host cells. In this study we investigated the benefits of division of labour for violacein production by splitting the violacein bioproduction pathway between two subpopulations of Escherichia coli based synthetic communities. We tested several pathway splitting strategies and reported that splitting the pathway into two subpopulations expressing VioABE and VioDC at a final composition of 60:40 yields a 2.5 fold increase in violacein production as compared to a monoculture. We demonstrated that the coculture outperforms the monoculture when both subpopulations exhibit similar metabolic burden levels, resulting in comparable growth rates, and when both subpopulations are present in sufficiently high proportions.

bioengineering↗

Enhancing Plant Photosynthesis using Carbon Dots as Light Converter and Photosensitizer

Improving photosynthetic efficiency is pivotal for CO2-based biomanufacturing and agriculture purposes. Despite the progress on photosynthetic biohybrids integrating biocatalysts with synthetic materials, nanomaterials with improved optical and photoelectrochemical properties are still needed to increase the energy-conversion efficiency. Here, we present a novel approach using carbon dots (CDs) as both intracellular photosensitizers and light converters for enhancing solar energy utilization in photosynthetic organisms. The CDs were produced from cyanobacterial biomass and used to convert a broad spectrum of solar irradiation to red light. We demonstrated that the nanosized CDs were incorporated into cyanobacterial cells and transferred light-excited electrons into the photosynthetic electron transfer chain. The biohybrids consisting of the CDs and Synechococcus elongatus exhibited increased growth rates, enhanced activities of both photosystems, and accelerated linear electron transport, compared with the cyanobacterial cells only. The supplementation of the CDs increased CO2-fixation rate and CO2-to-glycerol production by 2.4-fold and 2.2-fold, respectively. Furthermore, the CDs were shown to enhance photosynthesis and promote growth of Arabidopsis thaliana. The fresh weight of plant was increased 1.8-fold by CDs addition. These results reveal that simultaneous photosensitization and spectral modification could substantially improve the efficiency of natural photosynthesis. This study presents CDs as an attractive nanomaterial with great application potential in agriculture and solar-powered biomanufacturing.

synthetic biology↗

Bioconversion of Glucose-Rich Lignocellulosic Wood Hydrolysates to 3-Hydroxypropionic Acid and Succinic Acid using Engineered Saccharomyces cerevisiae

Background3-hydroxypropionic acid (3-HP) and succinic acid (SA) were announced as two of the top twelve value-added platform chemicals from biomass out of a group of over 300 potential compounds that could be made from biomass in a government report in 2004 (Werpy and Petersen, 2004) and in an updated report in 2010 (Bozell and Petersen, 2010). The screening criteria used in the report classified 3-HP and SA as direct petroleum replacement building block chemicals. 3-HP is a precursor to several high-value compounds, such as acrylic acid, 1,3-propanediol, acrylamide, and methyl acrylates, that ultimately end up in products such as fibers, contact lenses, diapers, fabric coatings, and other super absorbent polymers (SAPs). SA is a high-value platform chemical used in polyester production and a precursor for nylon and other bioplastics. Additionally, these reports identified pathways to building block compounds from sugars. Yeast fermentations were identified in these reports as a preferred potential pathway to 3-HP and SA production from sugars because of yeasts natural low pH tolerance. ResultsThe laboratory strain Saccharomyces cerevisiae BY4741 was engineered to produce either 3-HP or SA. These yeasts can convert fermentable sugars from glucose-rich lignocellulosic hardwood feedstocks into organic acid products such as 3-HP and SA under low pH conditions using exponential fed-batch cultivation strategies. Glucose-rich wood sugars provided a better growth environment for the engineered yeast strains, increasing production titers by 6.1 and 6.5 times for SA and 3-HP, respectively. ConclusionsThis study shows the potential of locally produced glucose-rich wood sugars to increase the production of platform chemicals necessary in the production of biobased polymers by engineered yeast cell factories.

microbiology↗

DELLA Proteins Recruit the Mediator Complex Subunit MED15 to Co-activate Transcription in Land Plants

DELLA proteins are negative regulators of the gibberellin response pathway in angiosperms, acting as central hubs that interact with hundreds of transcription factors and regulators to modulate their activities. While the mechanism of transcription factor sequestration by DELLAs to prevent DNA binding to downstream targets has been extensively documented, the mechanism that allows them to act as co-activators remains to be understood. Here, we demonstrate that DELLAs directly recruit the Mediator complex to specific loci in Arabidopsis, facilitating transcription. This recruitment involves DELLA amino-terminal domain and the conserved MED15 KIX domain. Accordingly, partial loss of MED15 function mainly disrupted processes known to rely on DELLA co-activation capacity; including cytokinin-dependent regulation of meristem function and skotomorphogenic response, gibberellin metabolism feedback, and flavonol production. We have also found that the single DELLA protein in the liverwort Marchantia polymorpha is capable of recruiting MpMED15 subunits, contributing to transcriptional co-activation. The conservation of Mediator-dependent transcriptional co-activation by DELLA between Arabidopsis and Marchantia implies that this mechanism is intrinsic to the emergence of DELLA in the last common ancestor of land plants. Significance StatementDELLA proteins are plant-specific transcriptional hubs integrating environmental signals with endogenous cues. In order to regulate downstream processes, DELLAs modulate the activity of hundreds of transcription factors and transcriptional regulators in various ways. Here, we describe the molecular mechanism underlying DELLA co-activator function. We show that DELLAs act as transcriptional activators in eukaryotic cells by interacting with the Mediator complex subunit MED15. Mediator function is necessary to regulate a subset of DELLA-regulated responses that are mediated by direct co-activation of DELLA-Transcription factors complexes. We further show that this mechanism is present in bryophyte DELLAs, and thus represents a conserved mechanism of DELLA function in land plants.

plant biology↗

Host-aware RNA-based control of synthetic microbial consortia

Microbial consortia have been utilised for centuries to produce fermented foods and have great potential in applications such as therapeutics, biomaterials, fertilisers, and biobased production. Working together, microbes become specialized and perform complex tasks more efficiently, strengthening both cooperation and stability of the microbial community. However, imbalanced proportions of microbial community members can lead to unoptimized and diminished yields in biotechnology. To address this, we developed a burden-aware RNA-based multicellular feedback control system that stabilises and tunes coculture compositions. The system consists of three modules: a quorum sensing-based communication module to provide information about the densities of cocultured strains, an RNA-based comparator module to compare the ratio of densities of both strains to a pre-set desired ratio, and a customisable growth module that relies either on heterologous gene expression or on CRISPRi knockdowns to tune growth rates. We demonstrated that heterologous expression burden could be used to stabilise composition in a two-member E. coli coculture. This is the first coculture composition controller that does not rely on toxins or syntrophy for growth regulation and uses RNA sequestration to stabilise and control coculture composition. This work provides a fundamental basis to explore burden-aware multicellular feedback control strategies for robust stabilisation of synthetic community compositions.

synthetic biology↗

Toward sustainable jet fuels: bioconversion of cellulose into isoprenoid biojet candidates using rumen bacteria and non-conventional yeast

In this study, organic acids were demonstrated as a promising carbon source for bisabolene production by the non-conventional yeast, Rhodosporidium toruloides, at microscale with a maximum titre of 1055 {+/-} 7 mg/L. A 125-fold scale-up of the optimal process, enhanced bisabolene titres 2.5-fold to 2606 mg/L. Implementation of a pH controlled organic acid feeding strategy at this scale lead to a further threefold improvement in bisabolene titre to 7758 mg/L, the highest reported microbial titre. Finally, a proof-of-concept sequential bioreactor approach was investigated. Firstly, the cellulolytic bacterium Ruminococcus flavefaciens was employed to ferment cellulose, yielding 4.2 g/L of organic acids. R. toruloides was subsequently cultivated in the resulting supernatant, producing 318 {+/-} 22 mg/L of bisabolene. This highlights the feasibility of a sequential bioprocess for the bioconversion of cellulose, into biojet fuel candidates. Future work will focus on enhancing organic acid yields and the use of real lignocellulosic feedstocks to further enhance bisabolene production. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=100 SRC="FIGDIR/small/500214v1_ufig1.gif" ALT="Figure 1"> View larger version (21K): org.highwire.dtl.DTLVardef@1662a84org.highwire.dtl.DTLVardef@717f66org.highwire.dtl.DTLVardef@1633b36org.highwire.dtl.DTLVardef@1c46c7a_HPS_FORMAT_FIGEXP M_FIG C_FIG

synthetic biology↗

A yeast-based tool for mammalian DGATs inhibitors screening

Dysregulation of lipid metabolism is associated with obesity, metabolic diseases but there is also increasing evidences of a relationship between lipid bodies (LBs) excess and some cancer. LBs synthesis requires diacylglycerol acyltransferases (DGATs) which catalyses the last step of triacylglycerol (TAG) synthesis, the main storage lipid form in lipid bodies. DGATs and in particular DGAT2, are therefore considered as potential therapeutic targets for the control of these pathologies. Here, the murine and the human DGAT2 were overexpressed in the oleaginous yeast Yarrowia lipolytica deleted for all DGATs activities, for evaluating the functionality of the enzymes in this heterologous host and to evaluate DGAT activity inhibitors. This work provide evidence that mammalian DGATs expressed in Y. lipolytica is a useful tool for screening chemical libraries to identify potential inhibitors or activators of these enzymes of therapeutic interest.

microbiology↗

Development of a dedicated Golden Gate Assembly platform (RtGGA) for Rhodotorula toruloides

Rhodotorula toruloides is a potential chassis to microbial cell factories as this yeast can metabolise different substrates into a diverse range of natural products, but the lack of efficient synthetic biology tools hinders its applicability. In this study, the modular, versatile and efficient Golden Gate DNA assembly system was adapted to the first basidiomycete, an oleaginous yeast R. toruloides (RtGGA). R. toruloides CCT 0783 was sequenced, and used for the RtGGA design. The DNA fragments were assembled with predesigned 4-nt overhangs and a library of standardized parts was created containing promoters, genes, terminators, insertional regions, and resistance genes. The library was combined to create cassettes for the characterization of promoters strength and to overexpress the carotenoid production pathway. A variety of reagents, plasmids, and strategies were used and the RtGGA proved to be robust. The RtGGA was used to build three versions of the carotenoid overexpression cassette were built by using different promoter combinations. The cassettes were transformed into R. toruloides and the three new strains were characterized. Total carotenoid concentration increased by 41%. The dedicated GGA platform fills a gap in the advanced genome engineering toolkit for R. toruloides, enabling the efficient design of complex metabolic pathways.

synthetic biology↗

A modular toolset for electrogenetics

Synthetic biology research and its industrial applications rely on the deterministic spatiotemporal control of gene expression. Recently, electrochemical control of gene expression has been demonstrated in electrogenetic systems (redox-responsive promoters used alongside redox inducers and an electrode), allowing for the direct integration of electronics with complex biological processes for a variety of new applications. However, the use of electrogenetic systems is limited by poor activity, tunability and standardisation. Here, we have developed a variety of genetic and electrochemical tools that facilitate the design and vastly improve the performance of electrogenetic systems. We developed a strong, unidirectional, redox-responsive promoter before deriving a mutant promoter library with a spectrum of strengths. We then constructed genetic circuits with these parts and demonstrated their activation by multiple classes of redox molecules. Finally, we demonstrated electrochemical activation of gene expression in aerobic conditions utilising a novel, modular bioelectrochemical device. This toolset provides researchers with all the elements needed to design and build optimised electrogenetic systems for specific applications.

synthetic biology↗

Production of human milk fat substitute by engineered strains of Yarrowia lipolytica

Human milk fat has a distinctive stereoisomeric structure where palmitic acid is esterified to the middle (sn-2) position on the glycerol backbone of the triacylglycerol and unsaturated fatty acids to the outer (sn-1/3) positions. This configuration allows for more efficient nutrient absorption in the infant gut. However, the fat used in most infant formulas originates from plants, which tend only to esterify palmitic acid to the sn-1/3 positions. Oleaginous yeasts provide an alternative source of lipids for human nutrition. However, these yeasts also exclude palmitic acid from the sn-2 position of their triacylglycerol. Here we show that Yarrowia lipolytica can be engineered to produce triacylglycerol with more than 60% of the palmitic acid in the sn-2 position, by expression of a lysophosphatidic acid phosphatase with palmitoyl-Coenzyme A specificity, such as LPAAT2 from Chlamydomonas reinhardtii. The engineered Y. lipolytica strains can be cultured on glycerol, glucose, palm oil or a mixture of substrates, under nitrogen limited condition, to produce triacylglycerol with a fatty acid composition that resembles human milk fat, in terms of the major molecular species; palmitic, oleic and linoleic acids. Culture on palm oil or a mixture of glucose and palm oil produced the highest lipid titre in shake flask culture and a triacylglycerol composition that is most similar with human milk fat. Our data show that an oleaginous yeast can be engineered to produce a human milk fat substitute ({beta}-palmitate), that could potentially be used as an ingredient in infant formulas.

bioengineering↗

Comparative genomics analysis of Chryseobacterium sp. KMC2 reveals metabolic pathways involved in keratinous utilization and natural product biosynthesis

Several efforts have been made to valorize keratinous materials, an abundant and renewable resource. Despite these attempts to valorize products generated from keratin hydrolysate, either via chemical or microbial conversion, they generally remain with an overall low value. In this study, a promising keratinolytic strain from the genus Chryseobacterium (Chryseobacterium sp. KMC2) was investigated using comparative genomic tools against publicly available reference genomes to reveal the metabolic potential for biosynthesis of valuable secondary metabolites. Genome and metabolic features of four species were compared, shows different gene numbers but similar functional categories. We successfully mined eleven different secondary metabolite gene clusters of interest from the four genomes, including five common ones shared across all genomes. Among the common metabolites, we identified gene clusters involved in biosynthesis of flexirubin-type pigment, microviridin, and siderophore, all showing remarkable conservation across the four genomes. Unique secondary metabolite gene clusters were also discovered, for example, ladderane from Chryseobacterium sp. KMC2. Additionally, this study provides a more comprehensive understanding of the potential metabolic pathways of keratin utilization in Chryseobacterium sp. KMC2, with the involvement of amino acid metabolism, TCA cycle, glycolysis/gluconeogenesis, propanoate metabolism, and sulfate reduction. This work uncovers the biosynthesis of secondary metabolite gene clusters from four keratinolytic Chryseobacterium spp. and shades lights on the keratinolytic potential of Chryseobacterium sp. KMC2 from a genome-mining perspective, providing alternatives to valorize keratinous materials into high-value natural products. ImportanceKeratin is an abundant and renewable resource from slaughterhouses or the poultry industry. Low-value products such as animal feed and fertilizer were generated from these feedstocks based on conventional processing like chemical conversion. In fact, microorganisms possess the potential to synthesize valuable natural products. In this work, we explored the metabolic potential of Chryseobacterium sp. KMC2, which was isolated with efficient keratinolytic capacity from a previous study. Comparative genomics analysis displayed similar functional categories against three publicly available reference genomes of keratin-degrading Chryseobacterium spp.. Eleven different secondary metabolite gene clusters of interest were mined among four genomes, including five common and unique ones. Furthermore, we provide a more comprehensive understanding of metabolic pathways on keratin utilization in Chryseobacterium sp. KMC2, with the involvement of amino acid assimilation and sulfate reduction. These findings contribute to expanding the application of Chryseobacterium sp. KMC2 on the valorization of keratinous materials.

microbiology↗

Drop-In Biofuel production by using fatty acid photodecarboxylase from Chlorella variabilis in the oleaginous yeast Yarrowia lipolytica.

BackgroundOleaginous yeasts are potent hosts for the renewable production of lipids and harbor great potential for derived products, such as biofuels. Several promising processes have been described that produce hydrocarbon drop-in biofuels based on fatty acid decarboxylation and fatty aldehyde decarbonylation. Unfortunately, besides fatty aldehyde toxicity and high reactivity, the most investigated enzyme, aldehyde-deformylating oxygenase, shows unfavorable catalytic properties which hindered high yields in previous metabolic engineering approaches.\n\nResultsTo demonstrate an alternative alkane production pathway for oleaginous yeasts, we describe the production of diesel-like, odd-chain alkanes and alkenes, by heterologously expressing a recently discovered light-driven oxidase from Chlorella variabilis (CvFAP) in Yarrowia lipolytica. Initial experiments showed that only strains engineered to have an increased pool of free fatty acids showed to be susceptible to sufficient decarboxylation. Providing these strains with glucose and light in a synthetic medium resulted in titers of 10.9 mg/L of hydrocarbons. Using custom 3D printed labware for lighting bioreactors, and an automated pulsed glycerol fed-batch strategy, intracellular titers of 58.7 mg/L were achieved.\n\nConclusionsOleaginous yeasts such as Yarrowia lipolytica can transform renewable resources such as glycerol into fatty acids and lipids. By heterologously expressing a fatty acid photodecarboxylase from the algae Chlorella variabilis hydrocarbons were produced in several scales from microwell plate to 400 ml bioreactors. The developed bioprocess shows a route to the renewable production of hydrocarbons for a variety of applications ranging from representing a substrate for further enzymatic or chemical modification or as a drop-in biofuel blend.\n\nShort abstractOleaginous yeasts are potent hosts for the renewable production of lipids, fatty acids and derived products such as biofuels. Here, we describe, the production of odd-numbered alkanes and alkenes with a length of 17 and 15 carbons by expression of a fatty acid photodecarboxylase (CvFAP) from Chlorella variabilis in different Yarrowia lipolytica strains under different regimes of blue light exposure in several scales from microwell plate to 400 ml bioreactors.

bioengineering↗