Digital Image Photometry of Xylose in Liquid Medium
Ma. Roselette L. Alojado-Rubianes1 and Ernesto J. del Rosario2*
1National Institute of Molecular Biology and Biotechnology (BIOTECH) and
2Institute of Chemistry, University of the Philippines Los Banos, Laguna, Philippines
*Corresponding author: This email address is being protected from spambots. You need JavaScript enabled to view it.
ABSTRACT
Digital image photometry (DIP) was used to determine xylose concentration in liquid medium. Xylose was quantified by its color reaction with phloroglucinol in a 96-well microplate sample holder using a commercial flat-bed scanner followed by image analysis with free-access software (ImageJ). The Red (R) values showed linear correlation versus xylose concentration in the range of 1.0–10.0 mg/mL with correlation coefficient of 0.987. The DIP method was found to be accurate based on experimental pentose (versus theoretical) concentration values. The observed limits of detection (LOD) and quantification (LOQ) for xylose were 0.65 mg/mL and 2.17 mg/mL, respectively. The DIP method gave results that are reliable and comparable to other technical protocols requiring expensive equipment such as high performance liquid chromatograph (HPLC) or UV-visible spectrophotometer. The DIP method was used to evaluate the extent of xylose assimilation in liquid fermentation medium by four yeast strains.
INTRODUCTION
Pentoses, especially xylose, have become attractive substrates for ethanol production because they can be obtained from pentosans, which are important components of agricultural and forestry raw materials. Only a handful of organisms are capable of converting xylose into ethanol, but with world-wide research efforts of molecular biologists and genetic engineers, recombinant isolates from E. coli, Z. mobilis, S. cerevisiae, and other microorganisms have been produced (Lin & Tanaka 2006). However, there is still a need for conventional screening research for wild type microorganisms that can convert xylose into ethanol. In both types of research, a rapid and reliable method is needed for determining xylose in fermentation medium – preferably using inexpensive equipment. . . . . read more
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