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With potential anti-tumor and antioxidant properties, the polysaccharide content of <i>D. nobile</i> is relatively lower than that of the other medicinal <i>Dendrobium.</i> To find high-content polysaccharide resources, the polysaccharide (DHPP-Ⅰs) was prepared from <i>D.</i> Second Love ‘Tokimeki’ (a <i>D. nobile</i> hybrid) and compared with DNPP-Ⅰs from <i>D. nobile</i>. DHPP-Is (Mn 31.09 kDa) and DNPP-Is (Mn 46.65 kDa) were found to be <i>O</i>-acetylated glucomannans (-Glc<i>p</i>-(1,4) and <i>O</i>-acetylated-D-Man<i>p</i>-(1,4) backbones), analogous to other <i>Dendrobium</i> polysaccharides. DHPP-Ⅰs had higher glucose content (31.1%) and a lower degree (0.16) of acetylation than DNPP-Ⅰs (15.8%, 0.28). Meanwhile, DHPP-Ⅰs and DNPP-Ⅰs had the same ability in the radical scavenging assay, which was milder than the control of Vc. Both DHPP-Is and DNPP-Is inhibited <i>SPC-A-1</i> cell proliferation in vitro, with obvious differences in dose concentrations (0.5–2.0 mg/mL) and treatment times (24–72 h). Therefore, the antioxidant activity of DHPP-Ⅰs and DNPP-Ⅰs is not associated with distinction in anti-proliferative activity. As a glucomannan derived from non-medicinal <i>Dendrobium</i>, DHPP-Ⅰs has similar bioactivity to other medicinal <i>Dendrobium,</i> and this could serve as a starting point for studying the conformational–bioactivity relationship of <i>Dendrobium</i> polysaccharides.