Laccase production from repeated batch cultures using free mycelia of Trametes sp. isolated in Korea was investigated. Among various carbon and nitrogen sources, glucose and ammonium tartrate showed the highest potential for laccase production. Kinetic parameters, mu(max) and K-s on the growth of free mycelia of Trametes sp. in the shake flask batch-culture were 0.472 day(-1) and 2.752 g/liter, respectively. Kinetic pattern of laccase product formation for free mycelia in the shake-flask batch culture was a type of mixed-growth associate production. The maximum laccase activity of immobilized mycelia in shake-flask batch cultures was 7,800 U/liter, whereas the laccase activity of free mycelia peaked at about 14,600 U/liter. Also, it was possible to produce laccase from repeated batch cultures of free mycelia in shake-flask that increased the laccase activity from 12,000 U/liter to 20,000 U/liter during 40 days culture time.