Microfluidization is a novel and effective technology to improve the properties of myofibrillar protein. The effects of microfluidization with varying pressures (0–120 MPa) on the physicochemical, structural and emulsifying properties of chicken myofibrillar protein (CMP) were studied. Microfluidization treatment remarkably increased the absolute ζ-potential, contact angle, solubility, emulsifying ability and emulsifying stability of CMP. Simultaneously, the turbidity of CMP decreased. After microfluidization treatment, more α-helix structures were transformed into disordered structures, more hydrophobic and negatively charged groups were exposed, leading to improvements in CMP properties. After 90 MPa treatment, the absolute ζ-potential, storage modulus, loss modulus and dynamic apparent viscosity of CMP-camellia oil emulsion reached the maximum values. The hydrophobic interaction between CMP and camellia oil induced CMP to expose more hydrophobic and negatively charged groups, leading to the improved emulsifying properties of CMP. Our results demonstrate that microfluidization treatment has great potential to improve the product qualities of emulsion-type meat products.