Biochemical properties of free and immobilized Candida viswanathii lipase on octyl-agarose support: Hydrolysis of triacylglycerol and soy lecithin

Abstract

A B S T R A C TMicrobial lipases are important enzymes in food and pharmaceutical industries. In this work, a Candida viswanathii lipase was purified by hydrophobic interaction chromatography on octyl Sepharose. The purification presented 78.4% yield and the enzyme was 8.7-fold purified, with specific activity 700.4 U/mg protein and 69 kDa molecular weight. Immobilization of the enzyme on the same support presented 72.5% yield and a derivative with 101% expressed activity (109.2 U/g support), indicating hyperactivation of the enzyme. Optimal activity for both free and immobilized lipase was observed at pH 4.0 and 45°C. The free and immobilized lipase showed broad-range stability from acid to neutral pH, and apparent activation and stability on organic solvents. The derivative was 60-fold thermostabilized in relation to the free enzyme and fully retained its activity after four cycles of p-nitrophenyl palmitate hydrolysis. Slight activation was observed with dithiothreitol and βmercaptoethanol. The free and immobilized lipase efficiently hydrolyzed monoesters, simple and mixed long chain triacylglycerols, as well as soy lecithin. The activity and stability in acid pH, the organic solvent tolerance and the lecithin hydrolysis indicate high potential application of the enzyme and its derivative in textile, food and pharma industries and for chemical synthesis.

Simultaneous high nutritional single cell oil and lipase production by Candida viswanathii

“…Enzyme activity assay. Lipase activity was assayed with p-nitrophenyl palmitate (pNPP) as a substrate (Almeida et al, 2018). pNPP was initially dissolved in 0.5 mL of dimethyl sulfoxide (DMSO), then diluted to 0.05 M with McIlvaine buffer pH 3.5, containing 0.5% Triton X-100 (w/v).…”

“…Almeida et al (2016) also presented the potential of C. viswanathii to produce lipase in a solid-state cultivation system using poultry fat as a carbon source. Purification and immobilization of C. viswanathii lipase produced in submerged conditions showed a potential application of this catalyst for the hydrolysis of triacylglycerols and phospholipids (Almeida et al, 2018). The accumulation of microbial oil by C. viswanathii was first studied by Ayadi et al (2016) as a potential single cell oil using agro-industrial waste.…”

A Valuable Product of Microbial Cell Factories: Microbial Lipase

“…Interestingly, a lipase isolated from Candida viswanathii exhibited its highest activity at pH of 4.0; at pH 2.0–3.0, the enzyme activity was reduced to 15–40%, although 80% and 90% of its activity was maintained at pH values of 3.5 and 4.5, respectively. At pH values > 6.5, the activity decreased although a second pH maximum at 8.0 was observed ( Almeida et al, 2018 ). However, a lipase from A. niger exhibited its highest activity at pH 4.0, with 90% of this activity maintained at higher pH values ( Zubiolo et al, 2014 ).…”

“…In the papermaking industry, the addition of lipase can remove only the contaminating resins and oils in the paper; this will serve to reduce the breakage and to ensure the paper quality and output ( Almeida et al, 2018 ).…”

LIP4 gene expression from Candida viswanathii strain

“…Biochemical properties of crude lipase showed optimum pH and temperature activities at 4.0 and 40 o C, high thermal stability and activity in organic solvent [8]. The purified lipase presented high solvent stability and hydrolytic activity on natural triacylglycerols and soy lecithin [9]. C. viswanathii was also able to produce high level of lipase using vegetable oils from Brazilian Amazonian with different fatty acids compositions [10].…”