The invention provides toothpaste containing a composite biological enzyme preparation and having multiple whitening effects and a preparation method of the toothpaste. The toothpaste comprises the following components in percentage by weight: 1%-10% of glycerol, 0.1%-3.0% of a thickening agent, 40%-80% of sorbitol (sugar) alcohol, 0.1%-0.5% of a sweetening agent, 0.02%-1.0% of an anticaries agent, 0.001%-0.5% of bromelain, 0.001%-0.5% of papain, 0.01%-2% of lysozyme, 0.01%-5% of polyphosphate, 0.1%-10% of pyrophosphate, 0.1%-1% of a preservative, 5%-50% of a grinding agent, 0.3%-4.0% of a foaming agent, 0.8%-2.0% of essence, 0.1%-1.0% of an alkaline pH value regulator, 0.1%-2.0% of an acidic pH value regulator and the balance of water. The pH value of the formula is controlled to be 6.0-8.0, under the enzymatic action of papain, bromelain and lysozyme, protein films and bacterial biofilms on the surfaces of teeth are subjected to enzymolysis, exogenous stains on the teeth are effectively removed to achieve the effect of whitening the teeth, and the problem of exogenous stains on the teeth caused by long-term diet is solved.