ISOLATION AND SELECTION OF ALKALINE PROTEOLYTIC BACTERIA FROM LEATHER PROCESSING WASTE AND ENZYME CHARACTERIZATION

Authors

  • BUDIASIH WAHYUNTARI
    tika@biotrop.org
    gency for the Assessment & Application of Technology, Indonesia
  • NISA R MUBARIK Agency for the Assessment & Application of Technology, Jakarta, Indonesia, Indonesia
  • MARITA ANGGARANi Bogor Agricultural University, Bogor, Indonesia, Indonesia
No. 22 (2004)
Research Paper
November 17, 2011

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ABSTRACT
The aims of this experiment were to isolate alkaline protease producing bacteria from leather processing waste,
and to study the biochemical properties of the enzyme produced by the selected bacteria.
Nine bacterial isolates incubated at 37"C, revealed proteolytic activity on skim milk containing media. Four
isolates were grown at pH 9 and another four isolates at pH 10 and only one isolate at pH 11. However, in further subculture,
there were only three isolates that showed proteolytic activity, namely, D2, D7, and Dl l .  Among the three isolates, isolate
D2 was the highest protease producer. The highest protease production (36.5U/L) was reached after a 36-hr fermentation at
pH 9.
The optimum activity of D2 protease was observed at pH 8 and 60"C. The enzyme was stable at pH range of 7-10, and at
temperature of 52-62"C. In the presence of 5mM EDTA or PMSF, the crude enzyme activity decreased to 7.04% and 23.29%
respectively, which indicated that the enzyme might be a metal dependent serine protease. Zymogram analysis revealed the
molecular weight of the enzyme was about 42.8kD.
Keywords:  leather/ waste/protease/alkaline