A Cold-Adapted and Organic Solvent-Tolerant Lipase from a Psychrotrophic Bacterium Pseudomonas sp. Strain YY31: Identification, Cloning, and Characterization

Abstract

A novel cold-adapted lipase (designated as LipYY31) was obtained from a psychrotrophic Pseudomonas sp. YY31. The strain YY31 was gram-negative, rod shaped, motile by means of one polar flagellum, and exhibited chemotaxis toward oil droplets under a microscope. The strain displayed remarkable degradation of edible oil and fat even at 5 °C. The LipYY31 DNA fragment contains an open reading frame of 1,410 bp which encoded a protein of 470 amino acids with an estimated molecular mass of 49,584 Da. LipYY31 showed high sequence similarity to those of subfamily Ι.3 lipase and had a conserved GXSXG motif around the catalytic Ser residue. Its optimal temperature was 25-30 °C, and it retained 20-40 % of its activity at 0-5 °C. The optimal pH value was 8.0. The activity was strongly inhibited by Cd(2+), Zn(2+), EDTA and was highly dependent on Ca(2+). Tricaprin and p-nitrophenyl caprate were the most favorable substrates among the triglycerides and p-nitrophenyl esters, respectively. LipYY31 also had high activity towards natural substrates including edible vegetable oils and animal fat. Furthermore, LipYY31 was very active and stable in the presence of several detergents and organic solvents. In particular, the lipase exhibited high stability against organic solvents such as methanol, ethanol, and isopropanol.

Features of pseudomonads growing at low temperatures: another facet of their versatility

“…Lipase‐mediated biodiesel production, which is normally carried out at 35–50°C, would benefit from substantial cost reductions if the process could be accomplished at lower temperatures. Several psychrotolerant and psychrophilic pseudomonads have been found that contain lipases showing high activity at low or moderate temperatures (Suzuki et al ., ; Luo et al ., ; Yamashiro et al ., , among others). Similarly, several cold‐adapted Pseudomonas strains secrete proteases that are active at low temperatures and might be of use in biotechnological applications (Margesin and Schinner, ; Hamamoto et al ., ; Aghajari et al ., ; Zeng et al ., ).…”

Identification, Characterization, and Immobilization of an Organic Solvent-Stable Alkaline Hydrolase (PA27) from Pseudomonas aeruginosa MH38

“…After an early study on an organic solvent stable lipase from Pseudomonas aeroginosa LST-03 [6], significant efforts have been done to identify novel organic solvent-stable enzymes from Pseudomonas aeruginosa , Stenotrophomonas maltophilia , and Bacillus sphaericus . To date, several organic solvent stable lipases were identified and characterized, including LipS [7], SML [8], LipYY31 [9], LC2-8 [10], PseA [11], and 205y lipase [12]. However, the number of organic solvent stable lipases is still limited, and the search for novel enzymes to satisfy the needs of practical applications is of high importance.…”

The production and characterization of a new active lipase from Acremonium alcalophilum using a plant bioreactor

“…These acetyl groups specifically can represent up to 4.32% of dry weight in a variety of crop species [28]. Deacetylation of aspen wood and wheat straw, with up to 90% acetyl ester removal, has indicated that as the xylan becomes increasingly deacetylated, it becomes 5–7 times more digestible [29]. Thus, the discovery and characterization of enzymes that can facilitate the decomposition of these polysaccharides could improve the viability of the cellulosic ethanol industry.…”