A trial for the suspects - bus driver Ram Singh, his brother Mukesh Singh, Pawan Gupta, Vinay Sharma and Akshay Thakur - is scheduled to begin on Tuesday 5 February in Saket District Court Complex in New Delhi.
All of the accused signed statements to say they were innocent of the 13 charges filed against them, which include gang rape, murder, kidnapping and conspiracy.
AP Singh, a lawyer defending two of the men, Vinay Sharma and Akshay Thakur, says Sharma was never on the bus and Thakur was hiding beneath a seat and never took part in the crime.
Another of the accused, Mukesh Singh, has replaced his lawyer, who claimed he was tortured in police custody. Singh no longer claims mistreatment.
The men filed into the court room with their faces covered, and the charges were read out to them. If found guilty, they could face the death penalty.
A sixth suspect will be tried separately in a juvenile court. If convicted, the maximum sentence he could receive is three years in a detention facility.
VK Anand, defence counsel for the brothers Ram Singh and Mukesh Singh, said: "All the five accused have pleaded not guilty.
"The charges being framed is one thing," Anand said, "but proving the charges is another."
The court will begin hearing evidence on 5 February, when prosecutors are expected to call three witnesses at the start of the trial.
Over the course of the trial, around 86 witnesses will be examined, according to the local news channel IBN Live.
Prosecutors say they have strong evidence against the five, including blood-stained clothing, DNA matches, mobile phone records, confessions and eye-witness statements.
Police say the gang lured the 23-year-old woman on to a bus in New Delhi, where they repeatedly raped and assaulted her with a metal bar before throwing her bleeding on to a highway.
She died of internal injuries two weeks after the attack, which took place on 16 December 2012.
The brutality of the attack provoked condemnation worldwide about sexual assaults against women.
Thousands of protesters took to the streets in the weeks that followed.
In response to the public outcry, on 1 February the Indian cabinet fast-tracked new, tougher penalties for sex crimes.
A commission under ex-chief justice JS Verma was appointed to examine the country's laws on sex crimes, issuing its report on 23 January.
Ministers approved many of the recommendations on Friday, paving the way for raising the penalty for gang rape to life.
Under the new rules, due to be signed into law by the president, gang rape that leads to death will be punishable by death.
Minimum penalties will be raised to 20 years for gang rape and rape of a minor. The laws will later be ratified by parliament.