Balakrishnan, who joined Infosys in 1991, was considered by some as the next chief executive officer after current CEO S D Shibulal retires in 2015.
Infosys has witnessed an exodus of senior-level executives since the company brought back founder N R Narayana Murthy to oversee operations in June.
Balakrishnan, was head of Infosys BPO, Finacle and India Business Unit Chairman of Infosys Lodestone. He had also served as the firm's chief financial officer earlier.
"V Balakrishnan has conveyed his intention to resign as a member of the board and from the services of the company. The resignation is effective December 31, 2013," Infosys said in a statement.
The company announced the induction of U B Pravin Rao as a full-time director and Kiran Mazumdar-Shaw, Chairman and Managing Director of Biocon, as an independent director.
Rao is currently a member of the Executive Council, Senior Vice President and Global Head-Retail, Consumer Packaged Goods, Logistics and Life Sciences.
Yesterday, Subrahmanyam Goparaju, senior vice president and a member of the executive council that frames business strategy, had resigned.
Goparaju, who joined the company in 1998, was also head of Infosys Labs -- the research and innovation division.
Ashok Vemuri, who led the company's operations in the Americas, resigned in August, the same month when Vice President and Financial Services Head for Americas Sudhir Chaturvedi quit.
In the previous month, Infosys Global Sales Head Basab Pradhan left while Stephen R Pratt, the firm's Head of Utilities and Resources for North America, had put in his papers last month.In September, Infosys Head of BPO sales in Australia Kartik Jayaraman and BPO Head Latin America Humberto Andrade had quit the company.