Tony Hackworth's side returned to action for the first time in two weeks and were hoping to pick up where they left off, but goals from Nathan Keetley, Luca DeFalco and Kieran Ryan reversed their last seven games. It was only his second loss. League match expedition.
Both teams squandered early chances in the first half, but neither goalkeeper was tested until Alfie Barnett made a fine low save just before the half-hour mark.
However, that did not alert Pickering as Keatley scored his 19th league goal of the season from eight yards out in the 38th minute.
The visitors nearly doubled their lead just before half-time with a quick counter-attack, but Barnett made a good jump out to clear the danger and still leave the Pikes with a chance.
Hackworth made two changes at half-time, bringing on Wayne Brooksby and Destiny Ifeananya for Suleimane Koulibaly and Alex Zacariou, but they again missed chances, including an indirect free-kick from six yards out. Passed by.
Instead, Penistone extended their lead on the hour, with DeFalco dancing his way into the box and finishing brilliantly past Barnett.
Omar Sanyang forced Christopher Snaith into two saves as Pickering tried to fight back, but it was the visitors who ended the game. In the dying minutes of the game, Sam Scrivens' shot was deflected towards Ryan, who calmly slotted home the goal.
Pickering will be hoping for a comeback when they welcome Rossington Main to Mill Lane on Saturday.
