The 16th Brigade was formed in October 1939, one of the first three infantry brigades of the Second AIF, and became part of the 6th Australian Division. Its designation was derived from the fact that there were already 15 infantry brigades in the Australian Military Forces, not, as is popularly believed, because there had been 15 infantry brigades in the original AIF.

 The 16th Brigade's four infantry battalions - 2/1st, 2/2nd, 2/3rd and 2/4th - were all recruited in New South Wales. In February 1941 the Second AIF was reorganised in accordance with the war establishments of the British Army. This resulted in the reduction of Australian infantry brigades from four to three battalions, and the 2/4th Battalion subsequently became part of the 19th Brigade. 

As a formation, the 16th Brigade fought in Libya, Greece, on the Kokoda Trail and Sanananda Track in Papua, and in the Aitape-Wewak campaign.

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