Pseudomonas aeruginosa BBRC-10036 was used for lipase production. The organism secreted the enzyme extracellulary. In order to purify the enzyme, precipitation was carried out first, and then this lipase was purified by Ion Exchange Chromatography leading to 2.3-fold purification and 11.47% recovery. Lipase from P.aeruginosa was entrapped into Ca-alginate gel beads and effect of independent variables such as alginate concentration (%w/v), CaCl2 concentration (M) and enzyme load (%v/v) on immobilization yield and activity of immobilized enzyme were investigated. Media optimization for immobilization of lipase was carried out by Response Surface Methodology. The optimum conditions were: sodium alginate concentration 2.5% (w/v), calcium chloride concentration 2.5 (M) and enzyme load 50% (v/v). Under these conditions, the highest immobilization yield and the optimum activity of immobilized enzyme obtained were 93.65% and 2.64 unit/g (IME), respectively.

Background: Lipase enzymes are of great importance in various industries. Currently, extensive efforts have been focused on exploring new lipase producer microorganism as well as genetic and protein engineering of available lipases to improve their functional features. Methods: For screening lipase-producing lactobacilli, isolated strains were inoculated onto tributyrin agar plates. Molecular identification of lipaseproducing Lactobacilli was performed by sequencing the 16Sr DNA gene, and a phylogenetic tree was constructed. The LAF_RS05195 gene, encoding lipase protein in L. fermentum isolates, was identified using specific primers, amplified by PCR (918 bp) and cloned into the pET28a (+) vector. The recombinant proteins were expressed 2, 4, 6, and 12 hours after induction with IPTG and assessed using the SDS-PAGE. Enzymatic activity of the purified recombinant protein was measured at 410 nm in the presence of ρ NPA and ρ-NPP. Results: Among five identified native lipase-producing isolates, one isolate showed 98% similarity with Enterococcus species. The other four isolates indicated 98% similarity to L. fermentum. After purification steps with NiNTA column, a single protein band of about 34 kDa was detected on SDS-PAGE gel. The enzymatic activity of purified recombinant protein alongside ρ-NPA and ρ-NPP was measured to be 0. 6 U/ml and 0. 2 U/ml, respectively. Conclusion: In the present research, a novel lipase/esterase from L. fermentum was cloned and expressed. The novel lipase/esterase has the merit to be further studied due to its substrate specificity.

Background and objectives: Industrial wastewater is worldwide health concern. Microorganisms present in the environment have an important role in the biodegradation of lipids, fats and proteins from wastewater. In this regard, microbial lipases and proteases are interesting research targets because of high stability, broad substrate specificity, high yields and availability. In this study, we analyze sequences encoding lipase of Pseudomonas putida and subtilisin of Bacillus subtilis for generation of a new recombinant protein for degradation of environmental contaminations caused by lipids and proteins. Methods: In this study, sequences of the genes encoding lipase and subtilisin were obtained from GenBank. To predict the 3D structure of the protein, modeling was carried out. The prediction of secondary structure, tertiary structure and solvent accessibility was carried using bioinformatics tools including I-TASSER, GoR4 and ExPasy. Results: The lipase-subtilisin fusion protein was wellcharacterized by bioinformatical studies with appropriate spatial and secondary structures. The protein had appropriate hydrophilicity, biological half-life and thermal and acidic stability. The codon optimization was performed appropriately. Conclusion: Overall, the bioinformatical analysis of the designed protein showed that the recombinant lipase-subtilisin protein has a stable structure both in vitro and in vivo, a negative normalized B-factor and lipolytic and proteolytic activities, which makes it suitable for treatment of lipid and protein contaminations.

Conjugated Linoleic Acid (CLA), Glycerol (G) and sunflower oil blends with varying concentration were subjected to enzymatic esterification using a 1, 3- specific immobilized lipase. CLA was used as acyl due to its purported health benefits. The transesterified lipids were evaluated for free fatty acids (FFA) and composition of fatty acids by gas chromatography. Lipozyme Rhizomucor mehei immoblized (RM IM) is preferred for enzymatic esterification because of more CLA incorporation and less free fatty acids at the end of reaction than Lipozyme Thermomyces lanuginosus (TL) IM. Response surface methodology (RSM) was used to determine the effects of three variables Glycerol concentration, reaction temperature, and amount of enzyme on the lipase catalyzed incorporation of CLA into Sunflower oil. The optimum condition for maximum CLA incorporation (96.7% Incorporation Yield), maximum reaction rate (0.0271 M/h) and minimum free fatty acids production (Acid No.2.73) was obtained, at 75oC by using 3 % enzyme and 0.27 M glycerol.