Organic solvent tolerant metallo protease of novel isolate Serratia marcescens PPB-26: production and characterization

Abstract

Proteases are a class of enzymes that catalyze hydrolysis of peptide bonds of proteins. In this study, 221 proteolytic bacterial isolates were obtained by enrichment culture method from soils of various regions of Himachal Pradesh, India. From these a hyper producer of protease was screened and identified by morphological and physiological testing and by 16S rDNA sequence as Serratia marcescens PPB-26. Statistical optimization of physiochemical parameters enhanced the protease production by 75 %. Protease of S. marcescens PPB-26 was classified as a metalloprotease. It showed optimal activity at 30 °C, pH 7.5 (0.15 M Tris–HCl buffer) and with 0.8 % substrate concentration. It had K m = 0.3 %, V max = 34.5 μmol min−1 mg−1 protein and a half life of 2 days at 30 °C. The enzyme was stable in most metal ions but showed increased activity with Fe2+ and Cu2+ while strong inhibition with Co2+ and Zn2+. Further investigation showed that the enzyme could not only retain its activity in various organic solvents but also showed increased activity with methanol and ethanol. The reported metalloprotease is thus a potential candidate for carrying out industrial peptide synthesis.

Purification and Characterization of a DegP-Type Protease from the Marine Bacterium Cobetia amphilecti KMM 296

“…The enzyme expectedly exhibited reduced activity in the presence of the serine protease inhibitor PMSF, suggesting that the CamSP protease belongs to the class of serine proteases (Table 4). Ethanol and isopropanol increased the activity of CamSP by an average of 4-times and glycerol and Triton-X-100 by 2-times, which are consistent with the data on some proteases, for example, a Serratia marcescens metalloprotease [25], making the C. amphilecti protease useful for industrial applications such as peptide synthesis [39,50].…”

“…Efforts are currently underway to find and isolate new bacteria from poorly understood habitats that can produce enzymes with unique properties suitable for industrial applications [ 25 ]. The World Ocean can become such a unique source of producers of new proteases.…”

Production of thermostable multiple enzymes from Bacillus amyloliquefaciens KUB29

“…and Mn 2+ increased the activity of their proteases. The increase of protease activity in the presence of divalent cations, as Fe 2+ and Cu 2+ , was also reported for other bacteria, such as Serratia marcescens PPB-26 (Thakur et al, 2016).…”

“…licheniformis K7A (Hadjidj et al, 2018) and B. licheniformis NH1 (Thakur et al, 2016), as well as proteases whose highest activity is obtained in acidic pH (Xu et al, 2015). The crude protease preparations obtained from B. licheniformis AE-2, B.…”

“…Cada bactéria produz enzimas com características particulares para atuarem em condições físico-químicas específicas. Por exemplo, algumas espécies como Arthrobacterarilaitensis (Siala R et al, 2015) e Serratiamarcescens (Thakur S et al, 2016) já foram relatadas por serem produtoras de proteases tolerantes a solventes orgânicos.…”