Beef is rich in protein and low in fat， which is liked by consumers. It is difficult to chew and has low added value. In order to improve the taste and flavor of beef， the effects of single or combination patented strains， including W9-3 Lactobacillus reuteri， M3-16 Lactobacillus paracasei and MY4 Kluyveromyces marxianus， on the physicochemicals and sensory qualities， biogenic amine formations and volatile flavor of beef were investigated. The results showed that the addition of starter cultures could significantly decreased the pH (below 5.5)， the water activities (decreased by 0.6%-4.1%)， TBA values and carbonyl values of beef compared with unfermented or naturally fermented beef. The addition of starter cultures could inhibit the formations of histamine， tryptamine， tyramine and butylenediamine， and at the same times， make the brightness and springiness of cooked beef increase， while the hardness， gumminess， chewiness and cohesiveness decrease. And the results of the combination groups were higher than single strain groups. Eighty-six volatile substances have been identified from roasted fermented beef by using SPME-GC-MS. The results of these flavor compounds relative contents and PCA analysis showed that the flavor of roasted beef after fermentation with starter cultures are different from that of unfermented and naturally fermented beef， with the formation of new alcohols， aldehydes and ketones. The sensory evaluations showed that the texture， color， odor and taste of beef could be improved by fermentation with starter culture. The starter， especially the compound starter， can improve the quality and the flavor， while inhibit the biogenic amine formation of fermented beef. This paper could offer the theoretical foundation to the beef fermentation technology optimation.