The UPSC Plant Cell Wall and Carbohydrate Analytical Facility is dedicated to plant cell wall researches, which is an important aspect in UPSC. Equip­ment and competence for applying a large range of standard methods have been established with various sample preparation equipment, permanently installed equipment for conventional wet chemical methods and state-of-the-art analytical instrument setups.

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Our competence for the analysis of lignocellulose and secondary xylem, besides fine detection of soluble sugars and starch in these materials, includes: carbohydrate composition, lignin composition, lignin and carbohydrate structures and molecular size distributions of poly­mers. Conventional wet chemistry methods such as Updegraff cellulose and Klason/thioglycolic acid/acetylbromide lignin have been optimized in order to reduce the amount of sample material required. The availability of parallel cell wall analytical methodologies for both model systems (Arabidopsis and Populus) facilitates and accelerates the research process with other plant species.

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DSC 2413The established methods include many dif­ferent protocols to analyze monosaccharide composition of a whole sample or fractionated cell walls. The instrumental backbone for many of those methods is gas chromatography/mass spectrometry or flame ionization detector (GC/MS(FID)). The facility offers two instruments for analytical pyrolysis-GC/MS (Py-GC/MS) that quickly yields highly reproducible and comprehensive chemical fingerprinting of carbohydrate and lignin types, while a third is permanently available for sugar analysis

DSC 2427 1For characterizing a wide variety of polymers, there is an analytical size exclusion chromatography (SEC) setup with a quadruple detector array for multi-angle laser light scattering (MALLS), viscometry, refrac­tive index (RI) and UV absorbance detection. Ion chromatography (IC) is also available for the analysis of monosaccharides without derivatization, as well as oligosac­charides that can be collected for further characterization.



Recently, increasing number of researchers related to bioenergy, renewable resources and novel biomaterials conducted experiments at the lab.

DSC 2396m 1Steering committee:
A steering committee (Totte Niittylä, Lorenz Gerber, Ewa Mellerowicz, Hannele Tuominen, Leif Jönsson, Junko Takahashi-Schmidt) oversees the work of the facility and decides what techniques should be developed.

Contacts:
Totte Niittylä (This email address is being protected from spambots. You need JavaScript enabled to view it.): Responsible scientist
Lorenz Gerber (This email address is being protected from spambots. You need JavaScript enabled to view it.): Researcher
Junko Takahashi-Schmidt (This email address is being protected from spambots. You need JavaScript enabled to view it.): Laboratory manager


Available analyses and the charge (from July 2016)

For UPSC associated groups

Type of analysis Charged per Annual fee payers (10,000 kr/year) Non annual fee payers
Soluble sugar (glucose, fructose and sucrose) 1 Elisa plate (96-well) 300 kr 600 kr
Starch 1 Elisa plate (96-well) 400 800
Updegraff cellulose (crystalline cellulose) 1 sample 40 80
Anthrone assay only 1 sample 20 40
Klason lignin (acid insoluble lignin) 1 sample 40 80
Thioglycolic acid (TGA) lignin 1 sample 40 80
Acetylbromide lignin 1 sample 40 80
Biphenyl assay for uronic acids 1 Elisa plate (96-well) 400 800
Total carbohydrate (Dubois) 1 sample 20 40
Hexose/6-deoxyhexose (Dische) 1 sample 20 40
Pyrolysis-GC/MS 1 sample 60 120
TMS sugars with GC/MS 1 sample 120 240
Alditol acetate sugars with GC/FID 1 sample 120 240
GC/FID or MS only 1 sample 60 120
TFA/monosaccharide with IC 1 sample 100 200
Apha-amylase (Roche) for AIR2 1ul 0,5 0,5

For External groups

Type of analysis Charged per Annual fee payers (10,000 kr/year) Non annual fee payers
Soluble sugar (glucose, fructose and sucrose) 1 Elisa plate (96-well) 300 kr 1200 kr
Starch 1 Elisa plate (96-well) 400 1600
Updegraff cellulose (crystalline cellulose) 1 sample 40 160
Anthrone assay only 1 sample 20 80
Klason lignin (acid insoluble lignin) 1 sample 40 160
Thioglycolic acid (TGA) lignin 1 sample 40 160
Acetylbromide lignin 1 sample 40 160
Biphenyl assay for uronic acids 1 Elisa plate (96-well) 400 1600
Total carbohydrate (Dubois) 1 sample 20 80
Hexose/6-deoxyhexose (Dische) 1 sample 20 80
Pyrolysis-GC/MS 1 sample 60 240
TMS sugars with GC/MS 1 sample 120 480
Alditol acetate sugars with GC/FID 1 sample 120 480
GC/FID or MS only 1 smaple 60 240
TFA/monosaccharide with IC 1 sample 100 400
Apha-amylase (Roche) for AIR2 1ul 0,5 1

Additional information

1. It is also possible to do: 1) preparation of alcohol insoluble residue (AIR) and removal of extractives, 2) analysis of acetylation/methyl esterification.
2. It is possible to do size exclusion chromatography (SEC) in collaboration with Leif Jönsson's group in the Department of Chemistry, Umeå University. Please contact us.
3. Method development fee
If you would like to try a new method with us, we can do so in the form of project. For starting up, we need to charge the method development fee for covering the cost of necessary materials.
Half year: 6 000 kr, 1 year: 11 000 kr
4. Sample preparation by a Cell Wall Lab staff
When you have a lack of lab workers, although you have a lot of samples to be analyzed in the cell wall lab…  We can give you a hand when it is needed with the additional sample preparation fee.
1h: 275 kr, 1 day (8h): 2 200 kr
Examples:
1) TMS sample preparation (ca. 10h)
2) TMS data processing & peak identification (ca. 2h)
3) Py-GC/MS extra processing & peak identification (ca. half day)
4) Py-GC/MS sample preparation
5) Cleaning ion source in MSD due to special samples


Selected publications including analyses in the facility:
  • Pawar PM-A, Derba-Maceluch M, Chong S-L, Gómez LD, Miedes E, Banasiak A, Ratke C, Gaertner C, Mouille G, McQueen-Mason SJ, Molina A, Sellstedt A, Tenkanen M and Mellerowicz EJ (2016). Expression of fungal acetyl xylan esterase in Arabidopsis thaliana improves saccharification of stem lignocellulose. Plant Biotechnology Journal 14: 387–397.
  • Mahboubi A, Linden P, Hedenström M, Moritz T and Niittylä T (2015). Carbon-13 tracking after 13CO2 supply revealed diurnal patterns of wood formation in aspen. Plant Physiology 168: 478-489.
  • Serk H, Gorzsás A, Tuominen H and Pesquet E (2015). Cooperative lignification of xylem tracheary elements. Plant Signal Behavior 10(4):e1003753. doi: 10.1080/15592324.2014.1003753.
  • Derba-Maceluch M, Awano T, Takahashi J, Lucenius J, Ratke C, Kontro I, Busse-Wicher M, Kosik O, Tanaka R, Winzéll A, Kallas Å, Lesniewska J, Berthold F, Immerzeel P, Teeri TT, Ezcurra I, Dupree P, Serimaa R, and Mellerowicz EJ (2015). Suppression of xylan transglycosylase PtxtXyn10A affects cellulose microfibril angle in secondary wall in aspen wood. New Phytologist 205: 666–681.
  • Gandla ML, Derba-Maceluch M, Liu X, Gerber L, Master ER, Mellerowicz EJ and Jönsson LJ (2015). Expression of a fungal glucuronoyl esterase in Populus: effects on wood properties and saccharification efficiency. Phytochemistry 112: 210–220.
  • Normark M, Pommer L, Gräsvik J, Hedenström M, Gorzsás A, Winestrand S and Jönsson LJ (2015). Biochemical conversion of torrefied Norway spruce after pretreatment with acid or ionic liquid. BioEnergy Research DOI 10.1007/s12155-015-9698-7.
  • Tolu J, Gerber L, Boily JF and Bindler R (2015). High-throughput characterization of sediment organic matter by pyrolysis–gas chromatography/mass spectrometry and multivariate curve resolution: A promising analytical tool in (paleo)limnology. Analytica Chimica Acta 880: 93–102.
  • Gerber L, Zhang B, Roach M, Rende U, Gorzsás A, Kumar M, Burgert I, Niittylä T and Sundberg B (2014). Deficient sucrose synthase activity in developing wood does not specifically affect cellulose biosynthesis, but causes an overall decrease in cell wall polymers. New Phytologist 203: 1220 – 1230.
  • Mahboubi A, Ratke C, Gorzsás A, Kumar M, Mellerowicz EJ and Niittylä T (2013). Aspen SUCROSE TRANSPORTER 3 allocates carbon into wood fibers. Plant Physiology 163: 1729-1740.
  • Pesquet E, Zhang B, Gorzsás A, Puhakainen T, Serk H, Escamez S, Barbier O, Gerber L, Courtois- Moreau C, Alatalo E, Paulin L, Kangasjärvi J, Sundberg B, Goffner D and Tuominen H (2013). Non-Cell-Autonomous Postmortem Lignification of Tracheary Elements in Zinnia elegans. Plant Cell 25(4): 1314-1328.
  • Roach M, Gerber L, Sandquist D, Gorzsas A, Hedenström M, Kumar M, Steinhauser MC, Feil R, Daniel G, Stitt M, Sundberg B and Niittylä T (2012). Fructokinase is required for carbon partitioning to cellulose in aspen wood. Plant Journal 70: 967 – 977.
  • Gerber L, Eliasson M, Trygg J, Moritz T and Sundberg B (2012). Multivariate curve resolution provides a high-throughput data processing pipeline for pyrolysis–gas chromatography/mass spectrometry. Journal of analytical and applied Pyrolysis 95: 95-100.
Facility Funding
The facility was initially funded by FuncFiber in order to enable basic cell wall chemical characterizations in its projects. Additional funding has been provided to maintain the instruments and to develop techniques in particular by UPSC Berzelii Centre, Spruce Genome Project (Knut and Alice Wallenberg Foundation), Bio4Energy, TC4F and Value Tree. Large numbers of samples have been analyzed in association with BioImprove projects.