She was the first submarine of the RNN manufactured from high-quality Steel 52, with the ability to dive at a depth of 80 metres (260 ft).
[4] De Rooij attributed the increase in speed and weight to his design, which was based on research done in Wageningen.
The inside of O 16 was also different from previous submarines of the RNN, hosting such amenities as a refrigerator and multiple sinks for the crew.
During their time in Washington, Commander van Waning and one of the guests aboard the submarine, F. A. Vening Meinesz were granted an audience with American President Franklin D.
The Spanish Civil War was taking place during this time, which led General Francisco Franco to block access of ships to the Mediterranean Sea with the aid of the Royal Italian Navy.
A Japanese attack was expected, while there were also rumors of German raiders who had their eyes on the Dutch East Indies.
In September 1940, O 16 and K XVIII were sent from Tanjung Priok to shadow the steamship Lematang and tanker Olivia, during their trip from Durban to Lourenço Marques, with the intention to sink any possible German raider.
Only in November did the Dutch government-in-exile in London decide that submarine division I, to which O 16 belonged, would come under British command.
[20] On 9 December, at 9 p.m., O 16 received a message from British command to sail for the coast of Siam together with other submarines in divisions I and II.
The following day, on 11 December at 6 a.m., the submarines were ordered to set course for the east coast of the Malay Peninsula, between Kota Bharu and Singora.
Earlier that night, however, O 16 had spotted a Japanese troopship and launched three torpedoes, but because of the bad weather the crew could not confirm if they had hit and sunk the ship.
[18] The bay was only 11 metres (36 ft) deep so O 16 had to stay on the surface while she approached the moored troopships using electric motors to make as little noise as possible.
[23][24] Nonetheless, the crew of O 16 held this achievement in high regard, and retreated to their home-port in Singapore with only one torpedo left.
On 15 December 1941, during her homebound voyage to Singapore, O 16 hit a Japanese naval mine near Tioman Island, while leaving the Gulf of Siam.
[25] The survivor, quartermaster Cor de Wolf, managed to swim to Dayang Island, and eventually got in contact with the Royal Netherlands Navy, which brought him to Singapore.
[28] They confirmed the identity of the ship, and an expedition was organized which included people from the navy, two newspapers, and two descendants of Commander Bussemaker.