Foreign Minister Penny Wong revealed the boost at the opening of the ASEAN-Australia Special Summit on Monday.
Security has been bolstered across the city and Victoria Police given special powers under the Terrorism Act as a precaution to protect dozens of leaders and dignitaries from the region.
Ms Wong revealed an additional $222.5 million will go towards water security, combating transnational crime and responding to climate change through the Mekong-Australia Partnership over five years.
An additional $40 million has also been allocated to expanding Australia's Southeast Asian maritime partnerships through skills development, training and technology sharing over four years.
Penny Wong says ASEAN nations have a shared responsibility to keep peace in the region. (Joel Carrett/AAP PHOTOS)
Ms Wong warned of a risk of conflict through "miscalculation" as she stressed ASEAN nations have a shared responsibility to keep peace in the region.
"What happens in the South China Sea, in the Taiwan Strait, in the Mekong subregion, across the Indo Pacific affects us all," Ms Wong told delegates.
Strengthening business connections and co-operation on maritime operations, climate change and clean energy are top of the agenda at the three day summit.
Newly elected President of the Philippines Ferdinand Marcos Jr, Malaysian Prime Minister Anwar Ibrahim and Vietnamese Prime Minister Pham Minh Chinh are among leaders attending the high-level talks hosted by Prime Minister Anthony Albanese.
Mr Albanese, who will meet with the Malaysian prime minister on Monday, said it was an opportunity to work with regional neighbours to create jobs in Australia.
"That's what our international engagements are about - benefit for Australia in terms of our economy, but also in our uncertain world and national security interests as well," he told reporters on Sunday.
Anthony Albanese says the ASEAN bloc is Australia's second-largest two-way trading partner. (Morgan Hancock/AAP PHOTOS)
The event is being held in Australia for the first time since 2018, offering leaders an opportunity to hold bilateral meetings on top of a packed program of panels and discussions across three days from Monday.
ASEAN, an abbreviation for the Association of Southeast Asian Nations, is made up of Indonesia, Malaysia, the Philippines, Vietnam, Brunei, Cambodia, Myanmar, Laos, Singapore and Thailand.
Australia is not a formal member but has been a dialogue partner for the past 50 years.
Timor-Leste will also observe the summit.
Security has been bolstered across Melbourne for the ASEAN-Australia Special Summit. (Joel Carrett/AAP PHOTOS)
Mr Albanese said ASEAN was Australia's second largest two-way trading partner when viewed as a bloc and noted more than one million Australians have Southeast Asian heritage.
"The Special Summit commemorates our shared history and focuses firmly on the future - on how we can deepen our ties and Australia's engagement with our region," Mr Albanese said."Australia sees ASEAN at the centre of a stable, peaceful, and prosperous region.
"Strengthening our relationship ensures our shared future prosperity and security."
Mr Albanese is due to meet privately with the leaders of 10 nations in addition to events held in public.