192.168.0.1:8000 = 2 192.168.0.10:80 = 1 192.168.0.100:3128 = 2 192.168.0.4:8080 192.168.0.5:7822 = 2 192.168.0.6:8005 searchValue = "192.168.0.10:80" newNumber = 2 f = open("datafile.txt", "r+b") for line in file: if line.split()[0] == searchValue: position = f.tell() - len(line) #the tell() method gives the current position in the file f.seek(position) f.write("%s = %d" % (searchValue, newNumber)) break f.close()