Small scale screening of yeast strains enables high-throughput evaluation of performance in lignocellulose hydrolysates

Laboratory automation is a key driver in biotechnology and an enabler for powerful new technologies and applications. In particular, in the field of personalized therapies, automation in research and production is a prerequisite for achieving cost efficiency and broad availability of tailored treatments. For this reason, we present the StemCellDiscovery, a fully automated robotic laboratory for the cultivation of human mesenchymal stem cells (hMSCs) in small scale and in parallel. While the system can handle different kinds of adherent cells, here, we focus on the cultivation of adipose-derived hMSCs. The StemCellDiscovery provides an in-line visual quality control for automated confluence estimation, which is realized by combining high-speed microscopy with deep learning-based image processing. We demonstrate the feasibility of the algorithm to detect hMSCs in culture at different densities and calculate confluences based on the resulting image. Furthermore, we show that the StemCellDiscovery is capable of expanding adipose-derived hMSCs in a fully automated manner using the confluence estimation algorithm. In order to estimate the system capacity under high-throughput conditions, we modeled the production environment in a simulation software. The simulations of the production process indicate that the robotic laboratory is capable of handling more than 95 cell culture plates per day.

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Bioreactor Co-Cultivation of High Lipid and Carotenoid Producing Yeast Rhodotorula kratochvilovae and Several Microalgae under Stress

Microorganisms ◽

10.3390/microorganisms9061160 ◽

2021 ◽

Vol 9 (6) ◽

pp. 1160

Author(s):

Martin Szotkowski ◽

Jiří Holub ◽

Samuel Šimanský ◽

Klára Hubačová ◽

Pavlína Sikorová ◽

...

Keyword(s):

Positive Impact ◽

Small Scale ◽

Biomass Growth ◽

Media Composition ◽

Yeast Strains ◽

High Lipid ◽

Algae Growth ◽

Chlorophyll Production ◽

Lipid Metabolites ◽

Present Pilot Study

The co-cultivation of red yeasts and microalgae works with the idea of the natural transport of gases. The microalgae produce oxygen, which stimulates yeast growth, while CO2 produced by yeast is beneficial for algae growth. Both microorganisms can then produce lipids. The present pilot study aimed to evaluate the ability of selected microalgae and carotenogenic yeast strains to grow and metabolize in co-culture. The effect of media composition on growth and metabolic activity of red yeast strains was assessed simultaneously with microalgae mixotrophy. Cultivation was transferred from small-scale co-cultivation in Erlenmeyer flasks to aerated bottles with different inoculation ratios and, finally, to a 3L bioreactor. Among red yeasts, the strain R. kratochvilovae CCY 20-2-26 was selected because of the highest biomass production on BBM medium. Glycerol is a more suitable carbon source in the BBM medium and urea was proposed as a compromise. From the tested microalgae, Desmodesmus sp. were found as the most suitable for co-cultivations with R. kratochvilovae. In all co-cultures, linear biomass growth was found (144 h), and the yield was in the range of 8.78–11.12 g/L of dry biomass. Lipids increased to a final value of 29.62–31.61%. The FA profile was quite stable with the UFA portion at about 80%. Around 1.98–2.49 mg/g CDW of carotenoids with torularhodine as the major pigment were produced, ubiquinone production reached 5.41–6.09 mg/g, and ergosterol yield was 6.69 mg/g. Chlorophyll production was very low at 2.11 mg/g. Pilot experiments have confirmed that carotenogenic yeasts and microalgae are capable of symbiotic co-existence with a positive impact om biomass growth and lipid metabolites yields.

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Torulaspora delbrueckii Yeast Strains for Small-scale Chenin blanc and Pinotage Vinifications

South African Journal of Enology and Viticulture ◽

10.21548/39-1-1652 ◽

2018 ◽

Vol 39 (1) ◽

Author(s):

V. van Breda ◽

N.P. Jolly ◽

M. Booyse ◽

J. van Wyk

Keyword(s):

Small Scale ◽

Torulaspora Delbrueckii ◽

Yeast Strains ◽

Chenin Blanc

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Equalizing growth in high-throughput small scale cultivations via precultures operated in fed-batch mode

Biotechnology and Bioengineering ◽

10.1002/bit.22349 ◽

2009 ◽

Vol 103 (6) ◽

pp. 1095-1102 ◽

Cited By ~ 24

Author(s):

Robert Huber ◽

Marco Scheidle ◽

Barbara Dittrich ◽

Doris Klee ◽

Jochen Büchs

Keyword(s):

High Throughput ◽

Small Scale ◽

Fed Batch ◽

Batch Mode

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Automated High-Throughput System Combining Small-Scale Synthesis with Bioassays and Reaction Screening

SLAS TECHNOLOGY Translating Life Sciences Innovation ◽

10.1177/24726303211047839 ◽

2021 ◽

pp. 247263032110478

Author(s):

Nicolás M. Morato ◽

MyPhuong T. Le ◽

Dylan T. Holden ◽

R. Graham Cooks

Keyword(s):

High Throughput ◽

High Throughput Screening ◽

Desorption Electrospray Ionization ◽

Ambient Ionization ◽

Machine Learning Algorithms ◽

Small Scale ◽

Label Free ◽

Minimal Sample ◽

Secondary Droplets ◽

Automated Platform

The Purdue Make It system is a unique automated platform capable of small-scale in situ synthesis, screening small-molecule reactions, and performing direct label-free bioassays. The platform is based on desorption electrospray ionization (DESI), an ambient ionization method that allows for minimal sample workup and is capable of accelerating reactions in secondary droplets, thus conferring unique advantages compared with other high-throughput screening technologies. By combining DESI with liquid handling robotics, the system achieves throughputs of more than 1 sample/s, handling up to 6144 samples in a single run. As little as 100 fmol/spot of analyte is required to perform both initial analysis by mass spectrometry (MS) and further MSn structural characterization. The data obtained are processed using custom software so that results are easily visualized as interactive heatmaps of reaction plates based on the peak intensities of m/ z values of interest. In this paper, we review the system’s capabilities as described in previous publications and demonstrate its utilization in two new high-throughput campaigns: (1) the screening of 188 unique combinatorial reactions (24 reaction types, 188 unique reaction mixtures) to determine reactivity trends and (2) label-free studies of the nicotinamide N-methyltransferase enzyme directly from the bioassay buffer. The system’s versatility holds promise for several future directions, including the collection of secondary droplets containing the products from successful reaction screening measurements, the development of machine learning algorithms using data collected from compound library screening, and the adaption of a variety of relevant bioassays to high-throughput MS.

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High throughput construction and small scale expression screening of multi-tag vectors in Escherichia coli

Methods ◽

10.1016/j.ymeth.2011.08.002 ◽

2011 ◽

Vol 55 (1) ◽

pp. 29-37 ◽

Cited By ~ 63

Author(s):

Louise E. Bird

Keyword(s):

Escherichia Coli ◽

High Throughput ◽

Small Scale ◽

Expression Screening

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A small scale in vitro system for high throughput gas production analysis – A comparison with the Hohenheim gas test

Animal Feed Science and Technology ◽

10.1016/j.anifeedsci.2018.04.001 ◽

2018 ◽

Vol 241 ◽

pp. 8-14 ◽

Cited By ~ 1

Author(s):

Karola Elberg ◽

Patrick Steuer ◽

Ute Habermann ◽

Jürgen Lenz ◽

Michael Nelles ◽

...

Keyword(s):

High Throughput ◽

Gas Production ◽

Small Scale ◽

In Vitro System ◽

Production Analysis

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Influence of media composition and temperature on volatile aroma production by various wine yeast strains

Czech Journal of Food Sciences ◽

10.17221/11/2008-cjfs ◽

2008 ◽

Vol 26 (No. 5) ◽

pp. 376-382 ◽

Cited By ~ 2

Author(s):

V. Petravić Tominac ◽

K. Kovačević Ganić ◽

D. Komes ◽

L. Gracin ◽

M. Banović ◽

...

Keyword(s):

Saccharomyces Cerevisiae ◽

Synthetic Medium ◽

Aroma Compounds ◽

Small Scale ◽

Wine Yeasts ◽

Wine Production ◽

Yeast Strains ◽

Aroma Production ◽

Volatile Aroma ◽

Volatile Aroma Compounds

Volatile aroma compounds production by two autochthonous Saccharomyces cerevisiae strains, isolated from Istria region, and three other yeast strains (Saccharomyces bayanus and two commercial Saccharomyces cerevisiae wine yeasts) was investigated on a small scale using synthetic VP4 medium and Graševina must at 12 and 20°C. The results obtained by gas chromatography analyses were compared with the aroma production properties of the native microflora, remaining after Graševina must sulphiting. In both media and at both temperatures, the wine yeasts investigated showed different metabolic profiles regarding the tested volatile aroma compounds, which should be taken in consideration for autochthonous wine production. Although the synthetic medium proved to be appropriate for the investigation of the fermentative properties, the determination of secondary aroma production by wine yeasts has to be conducted by must fermentation or possibly by fermentation of another synthetic medium whose composition would be more similar to must.

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