A ministry statement cited by the state news agency SANA said the border crossing was opened and the movement of trucks and transit to and from Syria resumed.
The border crossing was closed in 2015 when the rebels took over the Nasib area and the crossing in the countryside of Daraa province in Syria.
The Syrian Army captured Daraa and the border crossing in July after the rebels in the province surrendered and agreed to evacuate to rebel-held areas in northern Syria, mainly the northwestern province of Idlib.
Last week, the Transportation Ministry declared raising the transit crossing fees that should be paid by trucks passing in and out of Syria whether they were loaded or empty.
The Nasib crossing is the busiest border crossing in Syria as it is situated on the Damascus-Amman international highway.
The crossing is also considered one of the most important land crossings in the Middle East as it is the main crossing for Syrian exports to Jordan and the Gulf countries.
Lebanon would also benefit from the re-opening of the Nasib crossing as the small Mediterranean country has crossings with Syria only and through Nasib it can do business with Arab markets.
The crossing is important for Jordan which is suffering from economic problems. Since it was closed in 2015, the kingdom suffered heavy losses as 70 per cent of its exports and imports went through Syria, according to Nabil Ruman, head of the investor’s board of the Jordanian Free Zone.
Reports said the losses of the Jordanian transport sector from the border closure was estimated at around half a billion US dollars.