Addressing the growing global demand for food production, especially in the context of a 70% increase by 2050, necessitates innovative approaches to sustainable protein sources. This study focuses on harnessing the potential of sorghum proteins through a comparative analysis of two extraction methods—alkaline (NaOH) and NaOH-Ethanol-Reducing Agent (NER). The aim is to identify the technique with a higher extraction yield and subsequently characterize the extracted proteins. The NaOH method exhibited an extraction yield of 1%, equivalent to 7.2% of the total protein content, with a protein isolate content of 56.11%. In contrast, the NER method demonstrated a significantly higher extraction yield of 7.55%, constituting 58.1% of the total protein. The resulting sorghum protein isolate from the NER method displayed a remarkable protein content of 88.83%, differing significantly from the NaOH method. Amino acid composition analyses unveiled distinctive profiles. Glutamic acid, leucine, and alanine emerged as predominant amino acids in sorghum protein, while the NaOH-extracted proteins showcased elevated levels of glutamic acid, arginine, and leucine. Notably, the NER method did not exhibit amino acids Taurine and Hydroxyproline. Glutamic acid, leucine, and alanine remained prominent, accounting for 22.42%, 14.65%, and 9.43%, respectively. Selected for its higher extraction yield and protein purity, the NER method was further scrutinized for functional properties. Solubility analysis across pH levels revealed optimal solubility at pH 8 to 11, emphasizing potential applications in various pH-dependent formulations. Water Holding Capacity (WHC) was 178%, and Oil Holding Capacity (OHC) reached 424%, positioning sorghum proteins favorably compared to soybean and mung beans. This holistic exploration not only unveils the extraction nuances and amino acid intricacies of sorghum proteins but also sheds light on their functional prowess. The findings underscore sorghum’s viability as a protein source with diverse applications in the rapidly evolving landscape of sustainable food technology.