Comparative investigation of the macromolecular composition of mycelia forms Thielavia terrestris by infrared spectroscopy

Grūbe, Māra; Zagreba, E. D.; Gromozova, E. N.; Fomina, Marina

doi:10.1016/s0924-2031(98)00074-5

Cited by 51 publications

(24 citation statements)

References 18 publications

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“…A comparison of the RSD of the replicate spectra per sample to the RSD of the triplicate reference measurements showed that in general the IR method was more precise, indicating that most of the error with the model is due to errors in reference measurements. Grube et al (16) obtained similar results for quantification of major cell components. In this study we demonstrated the ability of FTIR spectroscopy in combination with multivariative statistics to quantitatively determine the PHB contents of bacterial cells, particularly recombinant E. coli cells.…”

mentioning

confidence: 63%

Quantitative Determination of the Biodegradable Polymer Poly(β-hydroxybutyrate) in a Recombinant Escherichia coli Strain by Use of Mid-Infrared Spectroscopy and Multivariative Statistics

Kansiz¹,

Billman‐Jacobe

McNaughton³

2000

Appl Environ Microbiol

133

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Fourier transform infrared (FTIR) spectroscopy in combination with the partial least squares (PLS) multivariative statistical technique was used for quantitative analysis of the poly(␤-hydroxybutyrate) (PHB) contents of bacterial cells. A total of 237 replicate spectra from 34 samples were obtained together with gas chromatography-determined reference PHB contents. Using the PLS regression, we were able to relate the infrared spectra to the reference PHB contents, and the correlation coefficient between the measured and predicted values for the optimal model with a standard error of prediction of 1.49% PHB was 0.988. With this technique, there are no solvent requirements, sample preparation is minimal and simple, and analysis time is greatly reduced; our results demonstrate the potential of FTIR spectroscopy as an alternative to the conventional methods used for analysis of PHB in bacterial cells.Many prokaryotic microorganisms synthesize polyhydroxyalkanoates (PHAs) as carbon and energy reserves (2,5,8,25,26), and poly(␤-hydroxybutyrate) (PHB) is the most common PHA. The general chemical structure of these compounds is shown in Fig. 1.The composition of the alkyl side chain (the R group) and the number of methylene units together determine the identity of the monomer. For PHB, the number of methylene units is 1 and the R group is CH 3 . A number of organisms have been developed in an effort to optimize the yield of PHB, and recently accumulations of PHB at levels up to 80% of dry cell weight have been obtained under optimum growth conditions (25). PHB is an attractive alternative to the environmentally unfriendly petrochemically derived plastics because its copolymer with polyhydroxyvalerate is a biodegradable molecule with properties similar to those of polyolefins, such as polypropylene and polyethylene. For this reason industrial fermentation of PHB has been the focus of a number of studies (1,38,39), while other studies have been directed towards the use of PHB-producing organisms in the breakdown of wastewater (32, 42).The most common method available at present for analysis of PHAs in bacterial cells is gas chromatography (GC) (6,22,31). The GC method involves hydrolysis and subsequent methanolysis or propanolysis of the PHAs in whole cells in the presence of sulfuric acid and chloroform. This method is timeconsuming and laborious and involves extensive use of solvents. Other methods of PHA analysis include gravimetry, infrared (IR) spectroscopy of chemically extracted PHB (23), fluorimetry (10), and cell carbon analysis (37). To aid in the development of more efficient fermentation processes and to monitor production, rapid feedback on the state of fermentation in terms of the PHA content of cells is required.Fourier transform infrared (FTIR) spectroscopy is a routine chemical technique used to study molecular structure, but when it is applied to a large collection of intact microbial cells, the resulting spectra reflect the total biochemical composition of the cells (28). IR spectroscopy can thus provid...

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mentioning

confidence: 63%

Quantitative Determination of the Biodegradable Polymer Poly(β-hydroxybutyrate) in a Recombinant Escherichia coli Strain by Use of Mid-Infrared Spectroscopy and Multivariative Statistics

Kansiz¹,

Billman‐Jacobe

McNaughton³

2000

Appl Environ Microbiol

133

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“…The common procedure of using band integrals or band ratios [20,[22][23] is critical due to peak overlay and different extinction coefficients of FTIR signals. Consequently, within this approach an iteration method was used, where FTIR spectra from standards of different macromolecules will be aligned to reconstruct the original cell spectra.…”

Section: Journal Of Biophotonicsmentioning

confidence: 99%

The use of FTIR spectroscopy to assess quantitative changes in the biochemical composition of microalgae

Wagner

Liu

Langner

et al. 2010

Journal of Biophotonics

122

104

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A mid-infrared spectroscopic method was developed for the simultaneous and quantitative determination of total protein, carbohydrate and lipid contents of microalgal cells. Based on a chemometric approach, measured FTIR (Fourier transform infrared) spectra from algal cells were reconstructed by a partial least square algorithm, using the spectra of the reference substances to determine their relative contribution to the overall cell spectrum. From this specific absorption, absolute macromolecular cell composition [pg cell(-1)] can be calculated using calibration curves, which have been validated by independent biochemical methods. The future potential of this method for photosynthesis research is shown by its application to follow time-resolved changes in the cellular composition of microalgae during an illumination period of several hours. We show how the macromolecular composition can be investigated by FTIR spectroscopy methods. This can substantially increase the efficiency of screening processes like bioreactor monitoring and may be beneficial in metabolic engineering of algal cells.

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“…In both applications, the biomass is obtained from the same microbiological source and results from different growth conditions. Grube et al (1999) established a multiple component quantitative analysis by infrared spectroscopy, based on the average molar extinction coefficients of the main cellular components at selected absorption bands. Since according to the Lambert-Beer law, the contribution of the individual components to a given absorption band is additive, the composition can be determined semi-quantitatively by multiple linear regression analysis.…”

Section: Introductionmentioning

confidence: 99%

“…Due to overlap of many bands in this region, a more detailed assignment cannot be made. A major peak around 1,080 cm À1 has been used before in quantitation of carbohydrates (Grube et al, 1999), but is not suitable as a general applicable method. For instance, around the same position the -PO 2 -symmetric stretching vibration in phospholipids and DNA is found and as an additional complication the Si-O stretching vibration in biomass resulting from diatoms also absorbs strongly around 1,075 cm À1 (Stehfest et al, 2005).…”

mentioning

confidence: 99%

Monitoring of biomass composition from microbiological sources by means of FT‐IR spectroscopy

Pistorius¹,

DeGrip²,

Egorova-Zachernyuk³

2009

Biotech & Bioengineering

153

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An FT-IR spectroscopic method was developed for the simultaneous quantitative analysis of biomacromolecular components in biomass, originating from various microbiological sources. For the determination of protein, lipid and carbohydrate content, creatine phosphokinase, egg phosphatidyl choline and starch hydrolysate were chosen as external standards. This selection was based on spectral similarity and ease of availability. Protein content was based on the area under the amide II band profile around 1,545 cm(-1). Because of the heterogeneous lipid composition in the different species, lipid content was determined using integration over the C-H stretching vibrational population between 2,984 and 2,780 cm(-1). Carbohydrate content was determined using integration over a C-O and C-O-C stretching band area between 1,180 and 1,133 cm(-1). Linear regression analysis provided three calibration lines, according to which biomasses from ten species were analyzed. This approach showed good intra-batch reproducibility. With this method we could demonstrate good reproducibility between batches of the same species with similar growth conditions while large differences in biomass composition were observed between the various species. Protein content as determined by FT-IR spectroscopy compared well with the results obtained from elemental analysis.

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Comparative investigation of the macromolecular composition of mycelia forms Thielavia terrestris by infrared spectroscopy

Cited by 51 publications

References 18 publications

Quantitative Determination of the Biodegradable Polymer Poly(β-hydroxybutyrate) in a Recombinant Escherichia coli Strain by Use of Mid-Infrared Spectroscopy and Multivariative Statistics

Quantitative Determination of the Biodegradable Polymer Poly(β-hydroxybutyrate) in a Recombinant Escherichia coli Strain by Use of Mid-Infrared Spectroscopy and Multivariative Statistics

The use of FTIR spectroscopy to assess quantitative changes in the biochemical composition of microalgae

Monitoring of biomass composition from microbiological sources by means of FT‐IR spectroscopy

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