Isolation and Production of Novel β-galactosidase from a Newly Isolated, Moderate Thermophile, Bacillus sp. Strain B1.1

Abstract

The enzyme β-galactosidase (lactase; EC 3.2.1.23) is a commercially important enzyme due to its various applications in dairy and food industries, which are based on the β-galactosidase-catalysed hydrolysis of lactose into glucose and galactose. The objectives of this work were to identify novel and attractive sources of this industrially relevant enzyme, and to study the effect of selected growth parameters (carbon source, lactose concentration, nitrogen source, peptone concentration, initial pH and temperature) on the formation of β-galactosidase. Based on a screening of isolates from Tha Pai hot spring, Mae Hong Son Province, Thailand, strain B1.1 was selected for further studies. Strain B1.1 is a Gram-positive, rod-shaped, catalase-positive bacterium that forms endospores. Based on the sequence of the 16S rDNA determined, this isolate is most closely related to Anoxybacillus sp. and Bacillus sp., and hence the strain is designated as Bacillus sp. B1.1. β-Galactosidase was produced by this strain with lactose and peptone as carbon and nitrogen sources, respectively. Optimal enzyme production occurred at an initial culture pH of 8.5 and at 45 °C. Under these optimum culture conditions, maximal volumetric and specific β-galactosidase activity of 0.478 U mL -1 and 0.338 U mg -1 protein, respectively, were obtained after 13 h of cultivation in a medium contain 2.5% lactose, 2.0% peptone, 0.3% K 2 HPO 4 , 0.1% KH 2 PO 4 and 0.05% MgSO 4 ·7H 2 O.