Sigamani Santhosh, Chinnasamy Ragavendran, Pannerselvam Balashanmugam, Natarajan Hemalatha, Ramamurthy Dhandapani
Center for Pharmaceutical Biotechnology, College of Medicine and Pharmacy, Duy Tan University, Da Nang, Vietnam.
Department of Cariology, Saveetha Dental College and Hospital, Saveetha Institute of Medical and Technical Sciences (SIMATS), Chennai, Tamil Nadu India.
3 Biotech. 2025 Jul;15(7):200. doi: 10.1007/s13205-025-04357-8. Epub 2025 Jun 2.
The present study focuses mainly on the extraction of C-Phycocyanin (CPC) pigment from cyanobacteria, its optimization and evaluation of its radical scavenging and antiproliferative abilities. Optimization of the pigment production was conducted both experimentally and statistically by response surface methodology (RSM). Quadratic models were fitted to the responses obtained from the box-Behnken design (BBD), to examine the interactions among Carbon, Nitrogen, Phosphate, pH and Temperature, and their impact on pigment production. Analysis of variance (ANOVA) validated the model,revealing that higher levels of carbon, nitrogen and phosphate significantly influenced ( < 0.05) CPC production. The optimum conditions were determined as Carbon-30 mM, Nitrogen-10 mM, phosphate-5.5 mM, Temperature-27.5 °C and pH-7.5 resulting in phycocyanin yield of 47.6 mg in 0.483g of cell mass. Under the RSM-optimized conditions, it was found that sp. was able to produce a twofold increase in CPC production compared to the BG 11 medium over a period of 34 days. A blue-colored pigment was extracted, with a total protein concentration of 278 µg/ml. Concentration of the pigment led to a tenfold increase in protein purity. Fourier transform infrared spectroscopy (FTIR) analysis of the lyophilized pigment and control phycocyanin revealed similar functional groups, including Hydroxyl group, C - H stretch, C = O stretch and C - N stretch. The molar mass of the CPC dimer was recognized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) as 19 kDa for α subunit and 21 kDa for β subunit. The free radical scavenging potential of the CPC was determined by IC values obtained from the DPPH = 56.45 μg/ml & ABTS = 85.8 μg/ml assays. Its cytotoxic potential was evaluated against HepG2 (liver), A549 (lung) & MCF7 (breast) cancer cell lines with an IC value recorded at 47.6, 47.7 and 63.2 µg/ml, respectively. Fluorescent imaging revealed apoptotic and necrotic stages in treated cancer cells, accompanied by cytochrome damage. DNA smearing was observed in HepG2 cells, indicating effective damage of genetic material. These findings suggests that CPC possesses promising antioxidant and anticancer potentials, making it a potential drug for pharmaceutical and food industries.
The online version contains supplementary material available at 10.1007/s13205-025-04357-8.
本研究主要聚焦于从蓝藻中提取藻蓝蛋白（CPC）色素，对其进行优化，并评估其自由基清除能力和抗增殖能力。通过响应面法（RSM）对色素生产进行了实验和统计优化。采用二次模型拟合从Box-Behnken设计（BBD）获得的响应，以研究碳、氮、磷、pH和温度之间的相互作用及其对色素生产的影响。方差分析（ANOVA）验证了该模型，结果表明较高水平的碳、氮和磷对CPC生产有显著影响（P＜0.05）。确定的最佳条件为：碳30 mM、氮10 mM、磷5.5 mM、温度27.5℃和pH 7.5，在0.483 g细胞质量中藻蓝蛋白产量为47.6 mg。在RSM优化条件下，发现该菌株在34天内CPC产量比BG 11培养基提高了两倍。提取了一种蓝色色素，总蛋白浓度为278 μg/ml。色素浓缩使蛋白纯度提高了10倍。对冻干色素和对照藻蓝蛋白进行傅里叶变换红外光谱（FTIR）分析，发现其具有相似的官能团，包括羟基、C-H伸缩振动、C=O伸缩振动和C-N伸缩振动。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）确定CPC二聚体的摩尔质量，α亚基为19 kDa，β亚基为21 kDa。通过DPPH（IC= 56.45 μg/ml）和ABTS（IC= 85.8 μg/ml）实验获得的IC值测定了CPC的自由基清除潜力。评估了其对HepG2（肝癌）、A549（肺癌）和MCF7（乳腺癌）癌细胞系的细胞毒性潜力，IC值分别为47.6、47.7和63.2 μg/ml。荧光成像显示处理后的癌细胞出现凋亡和坏死阶段，并伴有细胞色素损伤。在HepG2细胞中观察到DNA涂片，表明遗传物质受到有效损伤。这些发现表明CPC具有良好的抗氧化和抗癌潜力，使其成为制药和食品工业的潜在药物。
在线版本包含可在10.1007/s13205-025-04357-网站获取的补充材料。 8
