A Novel Cold-Adapted Lipase from Sorangium cellulosum Strain So0157-2: Gene Cloning, Expression, and Enzymatic Characterization

Abstract

Genome sequencing of cellulolytic myxobacterium Sorangium cellulosum reveals many open-reading frames (ORFs) encoding various degradation enzymes with low sequence similarity to those reported, but none of them has been characterized. In this paper, a predicted lipase gene (lipA) was cloned from S. cellulosum strain So0157-2 and characterized. lipA is 981-bp in size, encoding a polypeptide of 326 amino acids that contains the pentapeptide (GHSMG) and catalytic triad residues (Ser114, Asp250 and His284). Searching in the GenBank database shows that the LipA protein has only the 30% maximal identity to a human monoglyceride lipase. The novel lipA gene was expressed in Escherichia coli BL21 and the recombinant protein (r-LipA) was purified using Ni-NTA affinity chromatography. The enzyme hydrolyzed the p-nitrophenyl (pNP) esters of short or medium chain fatty acids (≤C10), and the maximal activity was on pNP acetate. The r- LipA is a cold-adapted lipase, with high enzymatic activity in a wide range of temperature and pH values. At 4 °C and 30 °C, the Km values of r-LipA on pNP acetate are 0.037 ± 0.001 and 0.174 ± 0.006 mM, respectively. Higher pH and temperature conditions promoted hydrolytic activity toward the pNP esters with longer chain fatty acids. Remarkably, this lipase retained much of its activity in the presence of commercial detergents and organic solvents. The results suggest that the r-LipA protein has some new characteristics potentially promising for industrial applications and S. cellulosum is an intriguing resource for lipase screening.

Cold and Hot Extremozymes: Industrial Relevance and Current Trends

“…In addition, Novozymes has developed two other alternative lipases: Lipex ® and Lipolase ® Ultra, which perform optimally in low–moderate temperatures. Research for the discovery and development of novel cold-adapted lipases from psychrophilic microorganisms is booming because of the useful application of lipases in detergents and the trend toward cold-washing detergency (Park et al, 2009 ; Xuezheng et al, 2010 ; Cheng et al, 2011 ; Litantra et al, 2013 ; Jiewei et al, 2014 ; Ji et al, 2015 ). A high-performance lipase from Pseudomonas stutzeri PS59 was recently identified for use in detergent formulations.…”

An Organic Solvent-Tolerant Alkaline Lipase from Cold-Adapted <i>Pseudomonas mandelii</i>: Cloning, Expression, and Characterization

“…[7][8][9] Here, we report the cloning, expression, and characterization of an organic solvent-tolerant alkaline lipase, LipS (GenBank ID JQ071496), from Pseudomonas mandelii JR-1. P. mandelii is a cold-adapted bacterium that grows at 4 C but not at 37 C, with an apparent optimum temperature range of 25-30 C. 10,11) To produce a clone, we utilized the genome sequence of Pseudomonas fluorescens Pf0-1, 12) which is closely related to P. mandelii, from among completed microbial genomes.…”

Expression and characterization of thermotolerant lipase with broad pH profiles isolated from an AntarcticPseudomonassp strain AMS3

“…In contrast, AMS3 lipase exhibited higher optimum activity compared to other reported lipases isolated from psychrophilic bacteria. r- LipA from Sorangium cellulosum ( Cheng et al, 2011 ), lipA1 from Psychrobacter sp. 7195 ( Zhang et al, 2009 ), lipase from Pseudomonas fragi strain IFO 3458 (PFL) ( Alquati et al, 2002 ) and lipX from Psychrobacter sp.…”