With today's action, the total seizure of diamonds, gold and jewellery, in this case, has gone up to Rs 56.49 billion.
The agency had said it arrived at this figure based on the stock value at showrooms and stores, which is also the price at which the company purchased the goods.
"ED conducted further searches at 35 locations across India in Nirav Modi case and seized diamonds, gold and jewellery worth Rs 549 crore (book value). Total seizure till now is Rs 56.49 billion," the Enforcement Directorate said in a statement.
The raids that began yesterday are expected to continue tomorrow, they said.
They added that both Modi and Choksi have been summoned under the Prevention of Money Laundering Act (PMLA) and asked to depose before the investigating officer of the case here on February 23.
These notices, they said, were handed over to the directors of the firms of the two businessmen as the two were not in the country. While Modi runs the jewellery brand chain under his name, Choksi is the promoter of Gitanjali Gems.
The ED had also moved the external affairs ministry to revoke their passports, following which the MEA suspended it temporarily today.
The agency has filed a fresh PMLA case, taking cognisance of a new CBI FIR, against Choksi and his promoted companies.
The agency had registered a PMLA case against the two and others based on a CBI FIR, which was the result of a PNB complaint.
The agency is probing if the allegedly defrauded bank funds were laundered and these proceeds of crime subsequently used by the accused to create illegal assets and black money.
Modi, 46, a regular feature on the lists of rich and famous Indians since 2013, was booked by the CBI, along with wife, brother and Choksi on January 31, for allegedly cheating the state-run PNB to the tune of Rs 2.80 billion.
The bank subsequently sent two more complaints to the CBI on Tuesday, saying the scam was worth more than Rs 114 billion.
The jewellery designer is understood to be a citizen of the country but his brother Nishal and wife Ami are not Indian nationals. They all left India between January 1 and 6, the CBI had said.