A Modular Liquid Sample Handling Robot for High-Throughput Fourier Transform Infrared Spectroscopy

Li, Jichun; Shapaval, Volha; Köhler, Achim; Talintyre, Robert; Schmitt, Jürgen; Stone, R.R.; Gallant, Andrew J.; Zeze, Dagou A.

doi:10.1007/978-3-319-23327-7_66

Cited by 13 publications

(17 citation statements)

References 13 publications

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“…Combined with the Duetz microtiter plate system (Duetz-MTPS), FTIR can serve as a rapid tool for monitoring of high throughput studies, such as screening of fungal strains for high and low value lipid production [10,70]. Moreover, high throughput screening was strengthen by a fully automated set-up of the biomass samples preparation for the FTIR-HTS analysis [71][72][73].…”

Section: Introductionmentioning

confidence: 99%

Microcultivation and FTIR spectroscopy-based screening revealed a nutrient-induced co-production of high-value metabolites in oleaginous Mucoromycota fungi

et al. 2020

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Mucoromycota fungi possess a versatile metabolism and can utilize various substrates for production of industrially important products, such as lipids, chitin/chitosan, polyphosphates, pigments, alcohols and organic acids. However, as far as commercialisation is concerned, establishing industrial biotechnological processes based on Mucoromycota fungi is still challenging due to the high production costs compared to the final product value. Therefore, the development of co-production concept is highly desired since more than one valuable product could be produced at the time and the process has a potentially higher viability. To develop such biotechnological strategy, we applied a high throughput approach consisting of micro-titre cultivation and FTIR spectroscopy. This approach allows single-step biochemical fingerprinting of either fungal biomass or growth media without tedious extraction of metabolites. The influence of two types of nitrogen sources and different levels of inorganic phosphorus on the co-production of lipids, chitin/chitosan and polyphosphates for nine different oleaginous Mucoromycota fungi was evaluated. FTIR analysis of biochemical composition of Mucoromycota fungi and biomass yield showed that variation in inorganic phosphorus had higher effect when inorganic nitrogen source-ammonium sulphate-was used. It was observed that: (1) Umbelopsis vinacea reached almost double biomass yield compared to other strains when yeast extract was used as nitrogen source while phosphorus limitation had little effect on the biomass yield; (2) Mucor circinelloides, Rhizopus stolonifer, Amylomyces rouxii, Absidia glauca and Lichtheimia corymbifera overproduced chitin/ chitosan under the low pH caused by the limitation of inorganic phosphorus; (3) Mucor circinelloides, Amylomyces rouxii, Rhizopus stolonifer and Absidia glauca were able to store polyphosphates in addition to lipids when high concentration of inorganic phosphorus was used; (4) the biomass and lipid yield of high-value lipid producers Mortierella alpina and Mortierella hyalina were significantly increased when high concentrations of inorganic phosphorus were combined with ammonium sulphate, while the same amount of inorganic phosphorus combined with yeast extract showed negative impact on the growth and lipid accumulation. FTIR spectroscopy revealed the co-production potential of several

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Section: Introductionmentioning

confidence: 99%

Microcultivation and FTIR spectroscopy-based screening revealed a nutrient-induced co-production of high-value metabolites in oleaginous Mucoromycota fungi

et al. 2020

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“…2100 cm −1 , could be utilized in peak normalization preprocessing [ 39 ]. In addition, we can expect that the use of a robotic system for sample preparation on silicone plates could increase the precision of HTS–FTIR measurement [ 31 ].…”

Section: Resultsmentioning

confidence: 99%

“…It has been shown that HTS FTIR spectroscopic analysis of lipids in cell biomass correlates very well with GC analysis, and can be used for the prediction of total lipid and several groups of fatty acids (saturated, monounsaturated, and polyunsaturated) in fungal cell biomass [ 7 ]. Analogous to Duetz MTPS, HTS FTIR features microplate design well suited for automation systems [ 31 ], thus it is far more suitable for high-throughput screening than probe- or cell-based ATR FTIR setting. In the present study we evaluate HTS FTIR spectroscopy as a unified method for simultaneous quantification of both intra- and extra-cellular metabolites, as well as substrate consumption in high-throughput screening of microbial bioprocesses.…”

Section: Introductionmentioning

confidence: 99%

FTIR spectroscopy as a unified method for simultaneous analysis of intra- and extracellular metabolites in high-throughput screening of microbial bioprocesses

et al. 2017

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BackgroundAnalyses of substrate and metabolites are often bottleneck activities in high-throughput screening of microbial bioprocesses. We have assessed Fourier transform infrared spectroscopy (FTIR), in combination with high throughput micro-bioreactors and multivariate statistical analyses, for analysis of metabolites in high-throughput screening of microbial bioprocesses. In our previous study, we have demonstrated that high-throughput (HTS) FTIR can be used for estimating content and composition of intracellular metabolites, namely triglyceride accumulation in oleaginous filamentous fungi. As a continuation of that research, in the present study HTS FTIR was evaluated as a unified method for simultaneous quantification of intra- and extracellular metabolites and substrate consumption. As a proof of concept, a high-throughput microcultivation of oleaginous filamentous fungi was conducted in order to monitor production of citric acid (extracellular metabolite) and triglyceride lipids (intracellular metabolites), as well as consumption of glucose in the cultivation medium.ResultsHTS FTIR analyses of supernatant samples was compared with an attenuated total reflection (ATR) FTIR, which is an established method for bioprocess monitoring. Glucose and citric acid content of growth media was quantified by high performance liquid chromatography (HPLC). Partial least square regression (PLSR) between HPLC glucose and citric acid data and the corresponding FTIR spectral data was used to set up calibration models. PLSR results for HTS measurements were very similar to the results obtained with ATR methodology, with high coefficients of determination (0.91–0.98) and low error values (4.9–8.6%) for both glucose and citric acid estimates.ConclusionsThe study has demonstrated that intra- and extracellular metabolites, as well as nutrients in the cultivation medium, can be monitored by a unified approach by HTS FTIR. The proof-of-concept study has validated that HTS FTIR, in combination with Duetz microtiter plate system and chemometrics, can be used for high throughput screening of microbial bioprocesses. It can be anticipated that the approach, demonstrated here on single-cell oil production by filamentous fungi, can find general application in screening studies of microbial bioprocesses, such as production of single-cell proteins, biopolymers, polysaccharides, carboxylic acids, and other type of metabolites.Electronic supplementary materialThe online version of this article (10.1186/s12934-017-0817-3) contains supplementary material, which is available to authorized users.

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“…However, the heterotrophic microalga C. cohnii reached a significantly higher biomass and lipid concentration in the MTPS than in the 1.5 L bioreactor, probably due to shear force sensitivity of this species. It is worth mentioning that the screening throughput of oleaginous microorganisms in the Duetz-MTPS can be increased by combining at-line FTIR spectroscopy and the automation of the cultivation-analytical system (Li et al 2016 ). Reproducibility and scalability results demonstrated that the Duetz-MTPS can be used for the cost-efficient, high-throughput screening of both single-cell and multicellular oleaginous microorganisms.…”

Section: Discussionmentioning

confidence: 99%

Assessment of the scalability of a microtiter plate system for screening of oleaginous microorganisms

Kósa

Vuoristo

Horn

et al. 2018

Appl Microbiol Biotechnol

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Recent developments in molecular biology and metabolic engineering have resulted in a large increase in the number of strains that need to be tested, positioning high-throughput screening of microorganisms as an important step in bioprocess development. Scalability is crucial for performing reliable screening of microorganisms. Most of the scalability studies from microplate screening systems to controlled stirred-tank bioreactors have been performed so far with unicellular microorganisms. We have compared cultivation of industrially relevant oleaginous filamentous fungi and microalga in a Duetz-microtiter plate system to benchtop and pre-pilot bioreactors. Maximal glucose consumption rate, biomass concentration, lipid content of the biomass, biomass, and lipid yield values showed good scalability for Mucor circinelloides (less than 20% differences) and Mortierella alpina (less than 30% differences) filamentous fungi. Maximal glucose consumption and biomass production rates were identical for Crypthecodinium cohnii in microtiter plate and benchtop bioreactor. Most likely due to shear stress sensitivity of this microalga in stirred bioreactor, biomass concentration and lipid content of biomass were significantly higher in the microtiter plate system than in the benchtop bioreactor. Still, fermentation results obtained in the Duetz-microtiter plate system for Crypthecodinium cohnii are encouraging compared to what has been reported in literature. Good reproducibility (coefficient of variation less than 15% for biomass growth, glucose consumption, lipid content, and pH) were achieved in the Duetz-microtiter plate system for Mucor circinelloides and Crypthecodinium cohnii. Mortierella alpina cultivation reproducibility might be improved with inoculation optimization. In conclusion, we have presented suitability of the Duetz-microtiter plate system for the reproducible, scalable, and cost-efficient high-throughput screening of oleaginous microorganisms.Electronic supplementary materialThe online version of this article (10.1007/s00253-018-8920-x) contains supplementary material, which is available to authorized users.

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A Modular Liquid Sample Handling Robot for High-Throughput Fourier Transform Infrared Spectroscopy

Cited by 13 publications

References 13 publications

Microcultivation and FTIR spectroscopy-based screening revealed a nutrient-induced co-production of high-value metabolites in oleaginous Mucoromycota fungi

Microcultivation and FTIR spectroscopy-based screening revealed a nutrient-induced co-production of high-value metabolites in oleaginous Mucoromycota fungi

FTIR spectroscopy as a unified method for simultaneous analysis of intra- and extracellular metabolites in high-throughput screening of microbial bioprocesses

Assessment of the scalability of a microtiter plate system for screening of oleaginous microorganisms

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