PMID- 24034438
OWN - NLM
STAT- PubMed-not-MEDLINE
DCOM- 20131115
LR  - 20211021
IS  - 1754-6834 (Print)
IS  - 1754-6834 (Electronic)
IS  - 1754-6834 (Linking)
VI  - 6
IP  - 1
DP  - 2013 Sep 14
TI  - Towards practical time-of-flight secondary ion mass spectrometry 
      lignocellulolytic enzyme assays.
PG  - 132
LID - 10.1186/1754-6834-6-132 [doi]
AB  - BACKGROUND: Time-of-Flight Secondary Ion Mass Spectrometry (ToF-SIMS) is a 
      surface sensitive mass spectrometry technique with potential strengths as a 
      method for detecting enzymatic activity on solid materials. In particular, 
      ToF-SIMS has been applied to detect the enzymatic degradation of woody 
      lignocellulose. Proof-of-principle experiments previously demonstrated the 
      detection of both lignin-degrading and cellulose-degrading enzymes on 
      solvent-extracted hardwood and softwood. However, these preliminary experiments 
      suffered from low sample throughput and were restricted to samples which had been 
      solvent-extracted in order to minimize the potential for mass interferences 
      between low molecular weight extractive compounds and polymeric lignocellulose 
      components. RESULTS: The present work introduces a new, higher-throughput method 
      for processing powdered wood samples for ToF-SIMS, meanwhile exploring likely 
      sources of sample contamination. Multivariate analysis (MVA) including Principal 
      Component Analysis (PCA) and Multivariate Curve Resolution (MCR) was regularly 
      used to check for sample contamination as well as to detect extractives and 
      enzyme activity. New data also demonstrates successful ToF-SIMS analysis of 
      unextracted samples, placing an emphasis on identifying the low-mass secondary 
      ion peaks related to extractives, revealing how extractives change previously 
      established peak ratios used to describe enzyme activity, and elucidating peak 
      intensity patterns for better detection of cellulase activity in the presence of 
      extractives. The sensitivity of ToF-SIMS to a range of cellulase doses is also 
      shown, along with preliminary experiments augmenting the cellulase cocktail with 
      other proteins. CONCLUSIONS: These new procedures increase the throughput of 
      sample preparation for ToF-SIMS analysis of lignocellulose and expand the 
      applications of the method to include unextracted lignocellulose. These are 
      important steps towards the practical use of ToF-SIMS as a tool to screen for 
      changes in plant composition, whether the transformation of the lignocellulose is 
      achieved through enzyme application, plant mutagenesis, or other treatments.
FAU - Goacher, Robyn E
AU  - Goacher RE
AD  - Department of Biochemistry, Chemistry and Physics, Niagara University, Niagara 
      University, NY, USA. rgoacher@niagara.edu.
FAU - Tsai, Alex Yi-Lin
AU  - Tsai AY
FAU - Master, Emma R
AU  - Master ER
LA  - eng
PT  - Journal Article
DEP - 20130914
PL  - England
TA  - Biotechnol Biofuels
JT  - Biotechnology for biofuels
JID - 101316935
PMC - PMC3847459
EDAT- 2013/09/17 06:00
MHDA- 2013/09/17 06:01
PMCR- 2013/09/14
CRDT- 2013/09/17 06:00
PHST- 2013/07/22 00:00 [received]
PHST- 2013/09/12 00:00 [accepted]
PHST- 2013/09/17 06:00 [entrez]
PHST- 2013/09/17 06:00 [pubmed]
PHST- 2013/09/17 06:01 [medline]
PHST- 2013/09/14 00:00 [pmc-release]
AID - 1754-6834-6-132 [pii]
AID - 10.1186/1754-6834-6-132 [doi]
PST - epublish
SO  - Biotechnol Biofuels. 2013 Sep 14;6(1):132. doi: 10.1186/1754-6834-6-132.