Sourdough bread is rich in nutrients, antioxidants, and distinctive flavor, contributing health benefits and extended shelf life. Therefore, the objective of this study was to compare the sourdough bread made by using different percentages (100 and 200g) of sourdough starter with and without addition of instant dry yeast (10g). All sourdoughs were fermented at room temperature for 6 hours and baked at 200°C for 25 minutes. The physicochemical analyses were then conducted on both the sourdough and bread. The nutritional profile of sourdough bread was evaluated. Additionally, the microbiological quality and sensory evaluation using a 9-point hedonic scale were also performed. Moreover, the shelf-life of sourdough bread was assessed over five days at room temperature. The results indicated that the fermentation with four conditions significantly reduced pH while increasing titratable acidity and lactic acid bacteria levels, with the values ranging from 4.43 to 5.63, 0.033 to 0.047%, and 6.16 to 9.45 Log CFU/g, respectively. In terms of color, the lightness (L*) decreased, while the redness (a*) and yellowness (b*) increased as more sourdough starter and yeast were added. For nutritional composition of sourdough bread, the carbohydrate, protein, fat, crude fiber and ash content were 38.70±1.14 to 44.74±0.35%, 11.8±50.14 to 12.37±0.09%, 2.75±0.07 to 2.79±0.02%, 1.42±0.14% to 1.7±0.31%, and 1.27±0.32 to 1.67±0.34%, respectively. For the result of sensory evaluation, the sourdough bread with 200g of sourdough starter and 10g of instant dry yeast provided a higher score for overall acceptability. And the moisture content and water activity showed no significant difference between them. For total plate count, and yeast and mold count, four formulated sourdough breads had no significant difference, ranging from 2.17 to 4.02 Log CFU/g, ND to 4.21 Log CFU/g, and ND to 4.03 Log CFU/g, respectively. And the shelf-life of sourdough bread with 200g of sourdough starter and 10g of instant dry yeast was 3 days at room temperature.