Novel thermostable lipase from Bacillus circulans IIIB153: comparison with the mesostable homologue at sequence and structure level

Abstract

Thermophilic Bacillus circulans IIIB153 isolated from hot springs of North West Himalayas, India, produced an extracellular lipase, which exhibited significant biofilm disruption property on the static biofilm disruption model with a single species of Actinomyces viscosous. The gene encoding the lipase was cloned and overexpressed in Escherichia coli. Recombinant Bacillus circulans lipase (BCL), a monomer with molecular mass of 43 kDa also exhibited significant biofilm disruption activity. The enzyme was optimally active at 60°C, pH 8.5 and retained >70% of its original activity after 1 h incubation at 60°C. 3D structure of BCL developed by homology modeling showed a typical α/β hydrolase fold, a characteristic feature of lipolytic enzymes. Comparison of thermostable BCL with mesostable lipase from Chromobacterium viscosum at the sequence and structure level showed distinct variations in the structural features, with the presence of a high content of proline residues, aromatic amino acids and salt bridges. These features along with the presence of zinc-binding site observed in BCL structure could have a potential role in thermal stability of the enzyme.

Isolation and characterization of a thermostable lipase from <i>Bacillus thermoamylovorans</i> NB501

“…The specific activity of Lip501r, under the standard condition of 50∞C in the presence of 5 mM CaCl 2 , was 414 units/mg. Similar calcium-dependent stabilization has been reported for a lipase from Bacillus circulans IIIB153 (Johri et al, 2012). A thermophilic lipase from Geobacillus stearothermophilus L1 has been reported to be bound to Zn 2+ (Kim et al, 1998).…”

Screening and characterization of a thermostable lipase from marineStreptomycessp. strain W007

“…• C [19]. Thus, lipase MAS1 was verified to be a thermostable enzyme based on the temperature profiles.…”

Molecular cloning and characterization of a thermostable lipase from deep-sea thermophile Geobacillus sp. EPT9

“…However, it is substantially more stable than lipases from Bacillus sp. L2 (about 20 % residual activity after 1 h at 70°C; Sabri et al 2009), Bacillus circulans IIIB153 (about 20 % residual activity of 1 h at 80°C; Johri et al 2012), Bacillus stearothermophilus L1 (about 12 % residual activity after 0.5 h at 70°C; Kim et al 1998), Bacillus stearothermophilus P1 (about 15 % residual activity after 1 h at 80°C; Sinchaikul et al 2001), Bacillus thermoleovorans ID-1 (about 25 % residual activity after 0.5 h at 80°C; Lee et al 2001), Geobacillus sp. T1 (no activity of 30 min at 80°C; Leow et al 2004), and Geobacillus sp.…”

“…In the past several years, a number of thermostable lipases have been characterized. These include lipases from Amycolatopsis mediterranei (Dheeman et al 2011), Aneurinibacillus thermoaerophilus (Masomian et al 2013), Bacillus circulans (Johri et al 2012), Bacillus stearothermophilus (Kambourova et al 2003;Kim et al 2000;Sinchaikul et al 2001), Bacillus subtilis (Olusesan et al 2011), Bacillus thermocatenulatus (Schmidt-Dannert et al 1997), Bacillus thermoleovorans (Castro-Ochoa et al 2005;Lee et al 1999), Bjerkandera adusta (Bancerz and Ginalska 2007), Burkholderia cepacia (Yang et al 2007), Caldanaerobacter subterraneus (Royter et al 2009), Geobacillus thermoleovorans (Abdel-Fattah and Gaballa 2008;QuintanaCastro et al 2009;Soliman et al 2007), Lactobacillus plantarum (Lopes et al 2002), Pseudomonas cepacia (Sugihara et al 1992), Staphylococcus aureus (Sarkar et al 2012), Streptomyces thermocarboxydus (H-Kittikun et al 2012), Thermoanaerobacter thermohydrosulfuricus (Royter et al 2009), Thermosyntropha lipolytica (Salameh and Wiegel 2007).…”