Humic acids have received a large attention of researching and development as amendments for poor fertility conditions and deteriorating physical properties, since the process of extracting is an easy common process to implement, as well as dilution of these acids with water enables to be used at economically acceptable levels over large areas. It is necessary to notice that the properties of these acids depending on the organic source and the concentration added to the soil.A laboratory experiment was conducted at the department of soil Sciences and water resources, collage of Agriculture ,University of Basrah ,Iraq to demonstrate the effect of humic acids extracted from different sources and inoculation with Bacillus subtilis or Aspergillus niger on alkaline phosphatase activity in calcareous soil calcareous, southern Iraq. Humic acids extracted from wheat straw, alfalfa leaves, goat manure, and poultry manure which composted previously . Humic acid levels were 0, 25 and 50 L ha-1 .The soil also inoculated with Bacillus subtilis or Aspergillus niger, then samples were incubated at 28±2°C. After 7 or 30 days of incubation the activity of alkaline phosphatase was estimated . Enzyme phosphatase activity was increased in soils treated with humic acid as compared to control with highest activity at Poultry manure (391.54 µg p-nitrophenol gm-1 soil.1h-1).The results also indicated that increasing the level of humic acid significantly increased enzyme activity. For soil treated with Bacillus subtilis and Aspergillus niger , phosphatase activity increased-as compared to control with a higher activity at Bacilluas subtilis (373.1 µg p-nitrophenol gm-1 soil.1h-1).