However, the investment details were not disclosed.
In February this year, Ant Financial had picked up stake in One97 Communications.
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The latest round of funding will allow Paytm to achieve scale and develop its vibrant mobile commerce and payment ecosystem in India and invest in marketing, technology and talent, One97 Communications said in a statement.
Investing in Paytm will enhance the ability of Alibaba and Ant Financial to tap opportunities in India's fast-growing mobile e-commerce marketplace and digital finance industry, it added.
"India is an important emerging market with strong e-commerce potential, and we look forward to partner with Paytm to deliver innovative products and services to consumers," Alibaba Group CEO Daniel Zhang said.
Alibaba, which is also an investor in e-commerce firm Snapdeal, has been looking at increasing investments in Indian companies.
Vijay Shekhar Sharma, Founder and Chief Executive Officer of Paytm, said, "Paytm is building India's most dominant mobile payment and commerce ecosystem.
"With the Alibaba and Ant Financial partnerships, we look to bring half a billion Indians to the mainstream economy and help millions of small businesses leverage this large m-commerce opportunity."
Citi acted as the exclusive financial advisor to Paytm on this transaction.
Paytm recently crossed 100 million wallet users who make over 75 million transactions every month.
The company has been aggressively investing in expanding its wallet, e-commerce and payment operations.
Paytm, which also has a payment bank licence, is also looking at investing heavily for rolling out its payment bank services.
Besides, it is also investing $10 million (approximately Rs 66 crore) in logistics data startup LogiNext.