Senior MP Elise Archer announced her departure from her ministerial roles, the party and politics on Friday afternoon.
"It has been my deep honour and privilege to serve my constituents in Clark and the people of Tasmania as attorney-general and in all my portfolios," she said in a statement.
"It is also my intention to notify her excellency of my resignation from the parliament of Tasmania as a member for Clark."
It was revealed on Thursday night Ms Archer faces an independent investigation into workplace bullying allegations.
Tasmania's government has been in minority since May, when two MPs quit the party to sit as independents.
The Liberals hold only 10 out of 25 seats in the state's lower house.
Premier Jeremy Rockliff held a snap press conference on Friday afternoon saying he had requested and accepted Ms Archer's resignation from cabinet earlier in the morning.
Ms Archer, who was elected to parliament in 2010, said it was clear to her the leadership of the Liberal party "continues to fail to support ambitious women".
"It takes incredible strength and resilience to be a woman in a senior government role and to strive to be treated the same as male colleagues, despite many years of dedicated, hardworking service," she said.
"I am particularly proud of my achievements as the first female Speaker … and significantly in law reform over the last five years."
She said political life had "taken a personal toll at times" but she hoped reform under her watch would leave an enduring legacy for Tasmania.
The Australian newspaper reported on Thursday night workplace complaints against Ms Archer had been raised by two current or former staff members with the state anti-discrimination commissioner.
The newspaper also reported Ms Archer criticised Mr Rockliff and his predecessor Peter Gutwein in text messages.
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