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The permeate (&lt;3 kDa), defined as the fraction of digested biogenic calcite. Cells remaining from the last step was further heated in 20 mL of 1% SDS containing 10 mM Tris solution (pH 6.8) at 95 \u00baC for 1 h. The supernatant was also collected through centrifugation and filtration, followed by desalting and concentration with 3 kDa Microsep centrifugal filter tubes. Subsequently, the pellet was further digested by 0.04 M NH2OH HCl + 4.35 M HAc mixture at 96 \u00baC for 6 h, with occasional agitation to obtain the intracellular Fe-Mn associated metabolitic biopolymer. The sum of these two fractions represents the intracellular biopolymers after cell breakage.</p>\n<p>Subsamples were taken from the concentrated biopolymers for the analysis of protein, total carbohydrate (TCHO) and uronic acid (URA), respectively. In brief, the protein abundance was measured through a modified Lowry protein assay, using bovine serum albumin (BSA) as the standard. 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