PARTIAL PURIFICATION AND CHARACTERIZATION OF THERMOSTABLE SERINE ALKALINE PROTEASE FROM Brevibacillus sp.

Sotharith PHON, Dr.rer.nat. Lucia Dhiantika Witasari, S. Farm., Apt., M. Biotech.

2021 | Tesis | MAGISTER ILMU DAN TEKNOLOGI PANGAN

The thermostable proteases that optimally withstand high-temperature condition from thermophilic bacteria could be produced and purified which are highly beneficial to the industry. Brevibacillus sp. isolated from Sikidang crater, Dieng plateau was counted as thermophilic bacteria which could grow at 70 ºC. The aim of this study was precisely to produce and partially purify and characterize the thermostable proteases produced from thermophilic Brevibacillus sp. The proteolytic index was found and protease was produced from Brevibacillus sp. and continuously purified by several crucial methods and followed by the enzyme characterization. Protease optimally produced from Brevibacillus sp. at 50 ºC and 18 h of incubation. The molecular weight of partial purified protease was 60 kDa. Partial purified thermostable proteases highly activated at 70 ºC, stable for 1h at pH 9.6, and inhibited by PMSF. Therefore this enzyme classified as alkaline serine protease. Moreover, this purified protease consisted of kinetic with Km of 1.94 mM and Vmax of 0.2 U/ml. Overall, alkaline serine protease from Brevibacillus sp. showed high activity at high temperature and alkaline pH which shall be vital for food industries like food bakery and brewing process.

The thermostable proteases that optimally withstand high-temperature condition from thermophilic bacteria could be produced and purified which are highly beneficial to the industry. Brevibacillus sp. isolated from Sikidang crater, Dieng plateau was counted as thermophilic bacteria which could grow at 70 ºC. The aim of this study was precisely to produce and partially purify and characterize the thermostable proteases produced from thermophilic Brevibacillus sp. The proteolytic index was found and protease was produced from Brevibacillus sp. and continuously purified by several crucial methods and followed by the enzyme characterization. Protease optimally produced from Brevibacillus sp. at 50 ºC and 18 h of incubation. The molecular weight of partial purified protease was 60 kDa. Partial purified thermostable proteases highly activated at 70 ºC, stable for 1h at pH 9.6, and inhibited by PMSF. Therefore this enzyme classified as alkaline serine protease. Moreover, this purified protease consisted of kinetic with Km of 1.94 mM and Vmax of 0.2 U/ml. Overall, alkaline serine protease from Brevibacillus sp. showed high activity at high temperature and alkaline pH which shall be vital for food industries like food bakery and brewing process.

Kata Kunci : thermostable enzyme, serine alkaline protease; Brevibacillus sp.; enzyme purification