Color and light are the main factors in the car manufacturing industry, with not much research, conducted on this topic, related to employee physical comfort. The factor affecting employee work comfort is the color factor and lighting time in their workplace environment. Color is the light, reflected by an object, then interpreted by the eye based on the light that hits the object. The value of the color wavelength in the spectrum of 380 nm to780 nm impacts worker performance. Good lighting will affect whether or not the room’s light conditions affect the illumination’s value. A good working room condition will affect the physical comfort of employees. Employee physical comfort has developed in many previous studies. However, research on the configuration of colors and lighting in the manufacturing industry’s work environment is still underdeveloped. This research aims to measure and determine the impact of color and lighting at specific color wavelengths on the illumination value and the employee’s physical comfort. It starts with studying the color, light, human comfort, then examines the effect on the manufacturing industry employees’ performance in the wiring installation assembly industry. Color and light settings are proven essential to get better human comfort. Green color with a wavelength of 490 nm to 570 can increase the highest illuminate average lux value of 12.43 %. Yellow can increase the heart rate by 4.85 %, while green can reduce Diastole by 2.95 % and Systole by 1.29 %. Pupillary size changes L by 23.17 % and R by 21.43 %. The effect of green color and lighting time can increase the value of illumination and decrease heart rate, blood pressure, and pupil size so that it impacts the physical comfort of employees.