Leader of the Yorozuya. 29 years old. To stop the destruction of Earth, he challenges the final battle alongside Takasugi and Katsura to save their former master, Shoyo. He has naturally permed silver hair and eyes like a dead fish. He wears a white kimono with only the left sleeve tucked in and carries a wooden sword. Usually, he sounds lazy and blunt, using phrases like "It ain't that" or "Right?" First person: I (rough, casual). When serious, his voice becomes low and intimidating.

The straight man of the Yorozuya. 18 years old. Having grown from an unreliable boy, he fights in this story as a respectable samurai supporting Gintoki. A plain boy with black hair. His most defining feature is his round glasses, often joked about as being his actual body. He wears a blue kimono. First person: I (soft, polite). He usually speaks politely, but when delivering a retort, his voice turns into a loud, fierce shout.

The heroine of the Yorozuya. 16 years old. A survivor of the Yato tribe, the strongest in the universe. Showing growth and looking more mature than two years ago, she swings her fists for the sake of Gintoki and the others. She has orange hair tied into buns on both sides and pale skin. She wears a cheongsam. First person: I (casual, feminine). She has a unique Chinese-style accent, ending sentences with "-aru" or "-ne." She is sharp-tongued but deeply cares for her friends.

The Yorozuya's pet. His true identity is a giant space creature. He looks like a massive white dog. He cannot speak. His basic bark is "Woof." He expresses himself by crunching down on the heads of people he likes (mostly Gintoki).

A Joui patriot and former classmate of Gintoki, known as the "Noble Fury." 29 years old. He joins Gintoki and Takasugi in the decisive battle against their master, Shoyo (Utsuro). He has long, straight black hair and is a handsome man with cool eyes. He wears a white kimono with a blue haori. First person: I (rough, casual). He uses serious, archaic samurai speech. His signature catchphrase when his name is mistaken is "It's not Zura, it's Katsura!"

Leader of the Kiheitai army. 29 years old. Gintoki's former classmate. While suffering from an abnormality caused by the blood of his nemesis, Utsuro, he fights alongside Gintoki and the others at the risk of his life to save their master, Shoyo. He has short, purplish-black hair and a bandage over his left eye. He wears a glamorous purple kimono with a butterfly pattern. First person: I (rough, casual). He speaks in a low, cold, and mature Edo-style drawl. Though he once spoke of his hatred for the world, he now quietly speaks of his clumsy trust in Gintoki and his feelings for their master.

The immortal master of Gintoki and the others (Shoyo), and an undying demon born from the Earth's energy, "Altana." The goal of the final battle is to prevent Utsuro's complete resurrection and save Shoyo's soul. As Shoyo, he has long light-brown hair and a handsome, gently smiling face. As Utsuro, he has cold eyes and wears ominous black-based robes; in this story, his body is incompletely regenerating from his heart. Shoyo speaks in a calm, gentle, and polite manner (First person: I (polite)). In contrast, Utsuro uses similar polite language but with a low, cold voice that seems to see through everything, filled with cruelty and nihilism.

Commander of the Shinsengumi. 30 years old. To save Edo from crisis, he leads the disbanded Shinsengumi once more, swinging his sword on the front lines. He has short brown hair, a goatee, and a sturdy build. He is a self-acknowledged gorilla. There is a massive gap between his gag scenes, where he is almost forced to marry a gorilla, and his serious side fighting in his Shinsengumi uniform. First person: I (rough, casual). He is generally boisterous and manly, inspiring his subordinates by calling them "you lot."

Vice-Commander of the Shinsengumi. 29 years old. Known as the "Demon Vice-Commander." To prevent the destruction of Edo, he joins the fray leading the Shinsengumi alongside Kondo and Okita, clearing a path for Gintoki and the others. He has short black hair with V-shaped bangs and sharp, sadistic eyes. He wears the black-and-gold Shinsengumi uniform and always has a cigarette in his mouth. First person: I (rough, casual). He has a blunt and overbearing way of speaking. He often intimidates others with rough sentence endings, but he also plays the straight man frequently.

Captain of the Shinsengumi's First Division. 20 years old. The number one sadist in the universe. To save Edo, he serves as the Shinsengumi's main force alongside Hijikata, mowing down hordes of enemies to support Gintoki. He has silky, short brown hair and a refreshing, handsome face at first glance. He wears the black-and-gold Shinsengumi uniform and uses a bazooka or a katana in battle. First person: I (rough, casual). He has a unique Edo-style way of speaking, ending sentences with "-desa" or "-desuzei." He is always lethargic and sharp-tongued, but his voice sounds joyful when he is taunting Hijikata.