Abstract
The amount, nature, and fate of DNA and protein in the major purification fractions generated during industrial scale refining of sugar cane into raw sugar by the diffuser and tandem roller mills was determined. The presence and size of sugar cane DNA were estimated using PCR and sugar cane specific primers that amplified fragments of various sizes from different segments of the repetitive intergenic region (IGS) of the 25S rDNA. Both the maximum fragment size capable of amplification and the amount of DNA decreased as refining progressed, indicating sequential degradation during the milling. However, PCR still detected minute quantities of sugar cane DNA in raw sugar (<10e-3 ppm). Using a bicinchoninic acid assay on trichloroacetic acid precipitated sodium dodecylsulfate-extracts, protein was found in all mill fractions and decreased from 4500 to 10 ppm as sugar cane was refined to raw sugar. Analysis of these extracts by one- and two-dimensional gel electrophoresis suggested a gradual degradation of proteins during refining. Shotgun proteomic analyses identified complex populations of sugar cane proteins, or peptides thereof, in all mill fractions, but the population complexity decreased during processing. Retail-purchased refined cane sugar showed no detectable protein or DNA (<2 ppm protein; 0.001 ppm DNA).
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