In Tajikistan, owning beef cattle is an important survival mechanism for smallholder farmers to alleviate poverty. Therefore, beef cattle farming enterprises should indeed strive to maximize profit to excel and flourish in a free economy. Nevertheless, smallholder beef cattle farmers are known for making little profit. Thus, this study was set to evaluate the profitability of beef cattle farming and its determinants to enhance profit maximization among smallholder beef cattle farmers in the Baljovan District of Khatlon region, Tajikistan. A total of 388 farming households were chosen at random and purposive for the study. The cross-sectional data collected using questionnaires was analyzed by using descriptive, gross margin (GM), and ordinary least squares (OLS) regression models. Based on the descriptive analyses, the mean age of beef cattle farmers was 52.73 years, with a household size of 7.07 members. The beef cattle farmers had an average of 18.23 cattle herd size with 8.54 years of farming experience. The average land area possessed by farmers was 10.59 hectares. Among farmers, men (98.2%) dominated beef cattle farming activities. Around 83.8% of farmers had a college grade (higher literacy). Besides, around 89.4% of farmers had access to farm credits. However, only 71.4% of farmers used farm credit points to produce beef cattle. Most of the farmers (89.7%) had access to accurate market information. Such market information enabled 75.8% of farmers to sell their beef cattle to open market (profitable) outlets rather than middlemen. About 89.4% had access to veterinary services. Additionally, about 82.7% of farmers acknowledged the availability of pasture for grazing, which motivated 87.6% of farmers to be involved in selling contracts. Furthermore, economic investigation results revealed that on average, farmers had a gross margin (GM-profit) of 353.77 US$ per cattle, with feed costs (58.6%) and medications costs (26.1%) accounting for the largest share of total variable costs. Meanwhile, the profitability of beef cattle farming among farmers was significantly influenced by education level, family size, farming experience, pasture availability, land size owned, selling contract, feed costs, medications expenses, access to credits, and sales costs (P < 0.05). This study concluded that beef cattle production is a feasible business. However, the potential for increased profitability is significant if existing resources are efficiently coordinated and production expenses, notably feed and healthcare costs, are minimized. Thus, the government should develop additional measures for addressing concerns such as capacity building, suitable and freely available pasture as well as health management, to boost beef cattle profitability among farmers in Tajikistan.