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To overcome these limitations, this study developed a novel glycerol\u2010based pretreatment strategy to disrupt the hydrogen bonding network of bacterial cellulose (BC), thereby improving its reactive accessibility. The results confirmed that glycerol pretreatment significantly enhanced the reactivity of BC. The reaction time for preparing carboxymethyl cellulose (CMC) from glycerol\u2010pretreated powdered BC was reduced from 2 to 1\u2009h, representing a 100% improvement in reaction efficiency. In addition, glycerol pretreatment facilitated the exposure of hydroxyl groups in BC, thereby increasing the degree of substitution of CMC from 0.297 to 0.433 and from 0.427 to a peak value of 0.626, an approximately 46% enhancement. In addition, the CMC films prepared from glycerol\u2010pretreated BC exhibited significantly improved light transmittance, reaching over 95% in the wavelength range of 300\u20131000\u2009nm. 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