Expression and Characterization of Coprothermobacter proteolyticus Alkaline Serine Protease

Abstract

A putative protease gene (aprE) from the thermophilic bacterium Coprothermobacter proteolyticus was cloned and expressed in Bacillus subtilis. The enzyme was determined to be a serine protease based on inhibition by PMSF. Biochemical characterization demonstrated that the enzyme had optimal activity under alkaline conditions (pH 8–10). In addition, the enzyme had an elevated optimum temperature (60°C). The protease was also stable in the presence of many surfactants and oxidant. Thus, the C. proteolyticus protease has potential applications in industries such as the detergent market.

Microbial population dynamics in temperature‐phased anaerobic digestion of municipal wastewater sludge

“…For instance, Proteiniphilum , Clostridium, Anaerobaculum , Coperothermobacter , Azorcus, Gelria, Sporanaerobacter, Coprococcus. Caldicoprobacter, Tepidanaerobacter , Paracoccus, Syntrophomonas , Gordonia and Acidovorax were the dominant genera shared between pyrotags in the sludge, and appeared to be well‐adapted to the infrastructure environment of anaerobic digesters . Members of the genera Clostridium , Coprococcus and Caldicobacter exhibit various features that demonstrate their potential for fermentation of a wide range of organic compounds and VFAs .…”

“…Caldicoprobacter, Tepidanaerobacter, Paracoccus, Syntrophomonas, Gordonia and Acidovorax were the dominant genera shared between pyrotags in the sludge, and appeared to be well-adapted to the infrastructure environment of anaerobic digesters. 23,60,63,64 Members of the genera Clostridium, Coprococcus and Caldicobacter exhibit various features that demonstrate their potential for fermentation of a wide range of organic compounds and VFAs. 57,60,63 Members of Coperothermobacter (Firmicutes) and Proteiniphilum (Bacteriodetes) are known as proteolytic bacteria, able to hydrolyze proteins to amino acids with proteases, and degrade amino acids to VFAs and NH 4 + N. 63,64 Gelria are strictly anaerobic, moderately thermophilic and obligately syntrophic, and play an important role in H 2 transfer to the methanogenic partners.…”

“…23,60,63,64 Members of the genera Clostridium, Coprococcus and Caldicobacter exhibit various features that demonstrate their potential for fermentation of a wide range of organic compounds and VFAs. 57,60,63 Members of Coperothermobacter (Firmicutes) and Proteiniphilum (Bacteriodetes) are known as proteolytic bacteria, able to hydrolyze proteins to amino acids with proteases, and degrade amino acids to VFAs and NH 4 + N. 63,64 Gelria are strictly anaerobic, moderately thermophilic and obligately syntrophic, and play an important role in H 2 transfer to the methanogenic partners. 65 The members of genus Syntrophomonas, represented as acetogenic syntrophic bacteria, were observed in all sludge samples but in comparatively larger amounts at 35 ∘ C. This genus utilizes long-chain fatty acids under low pressure of H 2 .…”

Microbial diversity in innovative mesophilic/thermophilic temperature-phased anaerobic digestion of sludge

“…The latter finding is in line with many previous studies where Coprothermobacter and Methanothermobacter were found to live in strict syntrophic associations (Sasaki et al 2011;Ge et al 2012). Proteolytic activity of Coprothermobacter is well documented (Etchebehere et al 1998;Tandishabo et al 2012;Majeed et al 2013;Lü et al 2014), and in particular this microorganism is capable to ferment proteins, growing well in presence of peptides (Ollivier et al 1985). Therefore, the solubilization of proteins into small molecules can promote Coprothermobacter establishment, especially at high temperatures (Lee et al 2009).…”