The invention discloses a preparation method of 3-methylol 
cefazolin. The preparation method comprises the following steps of dropwise adding a 7-ACA solution to a 1H-
tetrazole-1-
acetic acid anhydridemixing solution, performing a 
condensation reaction, after the reaction, adjusting pH 
phase splitting, then adding 
purified water, performing 
phase splitting, combining water phases, then adding an 
organic solvent for 
phase splitting, and performing concentration to obtain a primary 
purified water phase and a secondary 
purified water phase; and after adjusting pH of the secondary purified water phase, adding 
cephalosporin-C deacetylase, then adding a dispersing agent to the filtered water phase, then reducing the temperature, performing 
filtration after 
crystal formation, performing washing,and performing 
drying until the 
moisture is smaller than 1.0%. According to the preparation method disclosed by the invention, a traditional strong 
basic hydrolysis technology is substituted, waste 
water discharge can be reduced by 80%, the 
cephalosporin-C deacetylase after enzymolysis can be in cyclic utilization, 
solvent remains are smaller than 5%, the concentration weight 
reduction ratio is 2%-5%, the influence of various solvents on the enzymatic activity of the 
cephalosporin-C deacetylase is thoroughly eliminated, the consumption of enzymes and the activity of the enzymes being utilizedonce again are guaranteed, the cost is reduced, the reaction condition is mild, the conversion rate of products is as high as 90% or above, the purity of the products is not less than 98%, and subsequent structure elucidation and pharmacological research are facilitated.