Extracellular Lipase Research Articles

Engineering lipase TLL to improve its acid tolerance and its biosynthesis in Trichoderma reesei for biodiesel production from acidified oil

Lipases catalyze the synthesis of biodiesel, which is an important renewable alternative energy source. Cost-efficient conversion of waste acidified oil to biodiesel entails acid-tolerant lipases which have not been extensively studied. This study showed that the commonly used Thermomyces lanuginosus lipase TLL displayed a weak acid tolerance and an unsatisfactory performance in biodiesel production from acidified oil. Directed evolution of TLL identified one TLL-T3 variant with three residue substitutions (A69S/V150P/N222G). TLL-T3 displayed significantly enhanced acid tolerance, and its application in acidified oil treatment led to a biodiesel yield up to 90 % (w/w). A scaled-up production of TLL-T3 in Trichoderma reesei was further achieved and the highest extracellular lipase activity reached 16,123 U/mL after fermentation optimization. These results provide new insights into structural adaptation to acid tolerance by lipases and show that TLL-T3 holds great potential in commercial biodiesel production from waste acidified oil.

Evaluation of the potential probiotic Bacillus subtilis isolated from darkbarbel catfish (Pelteobagrus fulvidraco) on growth performance, serum immunity, and disease resistance of Aeromonas hydrophila.

This study aimed to identify potential probiotic strains of Bacillus subtilis from healthy fish gut microbiota for application in aquaculture. The effects of dietary B. subtilis administration on growth performance, serum enzyme activity, immune gene expression, and disease resistance in darkbarbel catfish (Pelteobagrus fulvidraco) were investigated. The isolate, identified through gene sequencing and biochemical tests, demonstrated resilience to pH 3.0% and 6.0% bile, and exhibited extracellular protease, cellulose, lipase, and amylase production. Darkbarbel catfish were fed diets with varying B. subtilis concentrations (0 CFU/kg [T0], 107 CFU/kg [T1], 108 CFU/kg [T2], and 109 CFU/kg [T3]). After 8 weeks, significant increases (p < 0.05) were observed in final body weight, weight gain rate, specific growth rate, serum lysozyme, serum superoxide dismutase, alkaline phosphatase, and total antioxidant capacity, whereas malondialdehyde levels significantly decreased. Feeding darkbarbel catfish with B. subtilis diets increased immunoglobulin M (IgM) and C3 gene expression (p < 0.05), indicating a positive impact on the fish's immune system. The strain upregulated interleukin 10 (IL-10) and transforming growth factor-β (TGF-β) expression and downregulated TNF-α and IL-1β, suggesting potential anti-inflammatory effects. Following a 7-day challenge with Aeromonas hydrophila, fish fed with B. subtilis exhibited lower mortality, with higher survival rates in the T2 and T3 groups. In conclusion, supplementing darkbarbel catfish diets with B. subtilis effectively enhances growth performance, immune response, and disease resistance.

Insights into the molecular mechanisms of lipid transformation in sturgeon fillets: Interplay between specific spoilage and dominant bacteria

This study investigates spoilage bacteria's impact on lipid metabolism in sturgeon fillets using UHPLC-Q-Orbitrap-MS/MS-based untargeted lipidomic analysis. A total of 4041 lipid molecules across five classes and 42 subclasses were identified, including glycerophospholipids (GPs, 50.88%), glycerolipids (GLs, 36.08%), sphingolipids (SPs, 10.47%), fatty acyls (FAs, 2.45%), and sterol lipids (STs, 0.12%). Aeromonas sobria, a specific spoilage bacterium, reduced GPs and FAs while increasing GLs, SPs, and STs via extracellular lipases and esterases. Acinetobacter albensis, the dominant bacterium, mainly elevated SPs and FAs. Their interaction promoted lipid metabolism and oxidation while producing volatile organic compounds (VOCs). Ethyl isobutyrate, ethyl propionate, isobutyl formate, pentan-2-one, propan-2-one, 2-butanone, 3-methyl-3-buten-1-ol, and dimethyl sulfide were mainly associated with Acinetobacter albensis, while 1-hexanol, 1-pentanol, 1-penten-3-ol, 1-hydroxypropan-2-one, 3-methyl-1-butanol, 2-methylbutanal, 3-hydroxy-2-butanone, and propionaldehyde were mainly related to Aeromonas sobria. This work unveils the mechanism of lipid transformation in sturgeon fillets during refrigerated storage, offering insights for aquatic products quality control.

Adsorption of extracellular lipase in a packed-bed reactor: an alternative immobilization approach.

In light of the growing demand for novel biocatalysts and enzyme production methods, this study aimed to evaluate the potential of Aspergillus tubingensis for producing lipase under submerged culture investigating the influence of culture time and inducer treatment. Moreover, this study also investigated conditions for the immobilization of A. tubingensis lipase by physical adsorption on styrene-divinylbenzene beads (Diaion HP-20), for these conditions to be applied to an alternative immobilization system with a packed-bed reactor. Furthermore, A. tubingensis lipase and its immobilized derivative were characterized in terms of their optimal ranges of pH and temperature. A. tubingensis was shown to be a good producer of lipase, obviating the need for inducer addition. The enzyme extract had a hydrolytic activity of 23 U mL-1 and achieved better performance in the pH range of 7.5 to 9.0 and in the temperature range of 20 to 50°C. The proposed immobilization system was effective, yielding an immobilized derivative with enhanced hydrolytic activity (35 U g-1), optimum activity over a broader pH range (5.6 to 8.4), and increased tolerance to high temperatures (40 to 60 ℃). This research represents a first step toward lipase production from A. tubingensis under a submerged culture and the development of an alternative immobilization system with a packed-bed reactor. The proposed system holds promise for saving time and resources in future industrial applications.

Novel extracellular lipase gene Lip1728 influences nutrient-dependent performance bacterial quorum sensing of Burkholderia pyrrocinia WZ10-3

Quorum sensing (QS) is a cellular communication mechanism in which bacteria secrete and recognize signaling molecules to regulate group behavior. Lipases provide energy for bacterial cell growth but it is unknown whether they influence nutrient-dependent QS by hydrolyzing substrate. A high-yield lipase-producing strain, Burkholderia pyrrocinia WZ10–3, was previously identified in our laboratory, but the composition of its crude enzymes was not elucidated. Here, we identified a key extracellular lipase, Lip1728, in WZ10–3, which accounts for 99 % of the extracellular lipase activity. Lip1728 prefers to hydrolyze triglycerides at sn-1,3 positions, with pNP-C16 being its optimal substrate. Lip1728 exhibited activity at pH 5.0–10.0 and regardless of the presence of metal ions. It had strong resistance to sodium dodecyl sulfate and short-chain alcohols and was activated by phenylmethanesulfonylfluoride (PMSF). Lip1728 knockout significantly affected lipid metabolism and biofilm formation in the presence of olive oil. Finally, oleic acid, a hydrolysate of Lip1728, influenced the production of the signal molecule N-acyl homoserine lactone (AHL) and biofilm formation by downregulating the AHL synthetase gene pyrI. In conclusion, Lip1728, as a key extracellular lipase in B. pyrrocinia WZ10–3, exhibits superior properties that make it suitable for biodiesel production and plays a crucial role in QS.

Properties of extracellular lipase from isolated bacterial on bovine milk fats

Properties of extracellular lipase from isolated bacterial on bovine milk fats

A new endophytic Penicillium oxalicum with aphicidal activity and its infection mechanism.

Aphid infestation adversely affects the yield and quality of crops. Rapid reproduction and insecticidal resistance have made controlling aphids in the field challenging. Therefore, the present study investigated the insecticidal property of Penicillium oxalicum (QLhf-1) and its mechanism of action against aphids, Hyalopterus arundimis Fabricius. Bioassay revealed that the control efficacy of the spores against aphids (86.30% and 89.05% on the third day and fifth day after infection, respectively) were higher than other components, such as the mycelium. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) showed that QLhf-1 invaded the aphid cuticle through spores and used the aphid tissues as a nutrient source for growth and reproduction, causing stiffness and atrophy and a final death. Three extracellular enzymes, lipase, protease, and chitinase had a synergistic effect with spores, and they acted together to complete the infection process by degrading the aphid body wall and accelerating the infection process. The newly discovered endophytic penicillin strain P. oxalicum 'QLhf-1' can effectively kill aphids. The results provided strong evidence for the biological control of aphids, and lay a foundation for the development and utilization of QLhf-1. © 2024 Society of Chemical Industry.

Biodegradation of synthetic plastics by the extracellular lipase of Aspergillus niger

A rapid increase in plastic pollution is a major threat to the environment. One intriguing group of enzymes that can act as biocatalysts for the breakdown of polymers is lipase. This study reports the production of lipase from Aspergillus niger MG654699.1 utilizing agro-industrial residue (wheat bran) through solid-state fermentation. The produced lipase showed 176.55 U/mL of activity, 7.18 mg/mL of protein content, and 24.60 U/mg of specific activity under the optimal conditions of 37°C and pH 7.0. The biocatalytic activity of 30 KDa lipase resulted in 3.8%, 3.6%, and 5% weight loss of PE (polyethylene), PET (polyethylene terephthalate), and PS (polystyrene), respectively. Application of Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM) confirmed the lipase-mediated deterioration of treated polymer samples. The alterations in functional groups and surface structures of the samples showed the chemical and physical impact of the applied enzyme. The findings of this study showed that lipase can be employed as an eco-friendly and green biocatalyst for effective depolymerization and deterioration of environmental plastic waste.

Effects of Olive Oil and Tween 80 on Production of Lipase by Yarrowia Yeast Strains

Lipase is one of the most commonly used biocatalysts in the food, pharmaceutical and cosmetic industries, and can be produced by Yarrowia lipolytica yeast. Despite the intensive studies of lipase from Yarrowia, there are still many open questions regarding the enzyme secretion process, especially by new isolates of this genus as well as the effect of substrates or surfactants, or both on the production of lipase. This research focused on the effect of olive oil and surfactant Tween 80 including the optimisation of the concentration of these compounds on the production of lipase by some novel Yarrowia isolates. Moreover, the optimal environmental parameters (pH, temperature) of crude enzyme synthetised by Yarrowia strains were determined. All investigated strains were able to produce lipase in both intracellular and extracellular fractions. The extracellular lipase activities were higher than the intracellular ones (Y. divulgata Y.02062 and Yarrowia lipolytica 854/4 147 U/L, 80 U/L and 474 U/L, 122 U/L, respectively). In the case of extracellular lipase, supplementing olive oil and Tween 80 enhanced significantly the synthesis and secretion of the enzyme. The lipase activity can even be enhanced by 20 times higher from 25 U/L to 474 U/L in the case of Yarrowia lipolytica 854/4 strain. In the case of intracellular, supplementation of Tween 80 generally reduces lipase activity except for the Y. lipolytica 1/4 strain, which was affected by two times the increase. The optimised concentration of olive oil and Tween 80 were determined for Y. divulgata Y.02062, Y. divulgata 5257, Y. lipolytica 1/4, and Yarrowia lipolytica 854/4 strains as 1.6% olive oil and 0.09% Tween 80, 1.6% olive oil and 0.06% Tween 80, 1.4% olive oil and 0.09% Tween 80 as well as 1.6% olive oil and 0.065% Tween 80, respectively. The optimum pH and temperature of crude lipases (intra and extracellular) synthetised by the tested Yarrowia lipolytica and Y. divulgata yeast strains were found to be pH 7.2 and 37 °C, respectively. Our results confirmed that the new isolate Y. divulgata is a very promising species for further development for industrial use as Y. lipolytica.

Exploring the Interplay between Nutrients, Bacteriophages, and Bacterial Lipases in Host- and Bacteria-mediated Pathogenesis.

Pathogenic bacteria and host cells counteract or neutralize each other's effect in two fundamental ways: Direct invasion and secretion of various substances. Among these, lipases secreted by pathogenic bacteria and host cell lysozyme are key actors. Secreted lipases from pathogenic bacterial are suggested as a key player in the pathogen-host interaction. Among the gut microbial energy sources, glucose and fats have been referred to as one of the best inducers and substrates for bacterial lipases. Enrichment of bacterial growth medium with extra glucose or oil has been shown to induce lipase production in pathogenic bacteria. More recently, research has focused on the role of human gut phage alterations in the onset of dysbiosis because the bacteria-phage interactions can be dramatically affected by the nutrient milieu of the gut. However, the reciprocal role of bacterial lipases and phages in this context has not been well studied and there is no data available about how high glucose or fat availability might modulate the cellular milieu of the pathogenic bacteria-phageeukaryotic host cell interface. The purpose of this study was to evaluate the immunologic outcome of pathogenic bacteria-phage interaction under normal, high glucose, and high butter oil conditions to understand how nutrient availability affects lipase activity in pathogenic bacteria and, ultimately, the eukaryotic host cell responses to pathogenic bacteria-phage interaction. 10 groups of co-cultured T84 and HepG2 cells were treated with Pseudomonas aeruginosa strain PAO1 (P.a PAO1) in the presence and absence of its KPP22 phage and incubated in three different growth media (DMEM, DMEM + glucose and DMEM + butter oil). Structural and physiological (barrier function and cell viability), inflammatory (IL-6 and IL-8), metabolic (glucose and triglycerides), and enzymatic (lipases and lysozyme) parameters were determined. Excess glucose or butter oil enhanced additively extracellular lipase activity of P.a PAO1. Excess glucose or butter oil treatments also magnified P. a PAO1- induced secretion of inflammatory signal molecules (IL-1β, IL-6) from co-cultured cells, concomitant with the enhancement of intracellular triglycerides in co-cultured HepG2 cells, these effects being abolished by phage KPP22. The results of the present study imply that KPP22 phage influences the interplay between food substances, gut bacterial lipases, and the gut cellular milieu. This can be applied in two-way interaction: by affecting the microbial uptake of excess free simple sugars and fats from the gut milieu leading to decreased bacterial lipases and by modulating the immune system of the intestinal -liver axis cells. Further studies are needed to see if the biological consequences of these effects also occur in vivo.

Bioprospecting Antibacterial Properties of Fungi Isolated from Kakinada Sea Coast and Eucalyptus Foliage

Background: Fungi produce a wide range of secondary metabolites with bactericidal or bacteriostatic properties. In search of novel antibacterial compounds recently, many fungi of marine and plant origin have been studied for their antimicrobial properties. Aims: This work aimed to study bio-prospect marine and endophytic fungi for their antibacterial properties. objective: To determine the antibacterial properties of ethyl acetate extracts of fungal isolates. Methods: Intensive microbiological methods were followed for isolation, differential growth, and qualitative screening of enzyme production. The isolates were characterised and identified based on morpho-taxonomy, 18S rRNA gene sequencing, and phylogenetic analysis of the tar-get genes. The antimicrobial activity of fungal ethyl acetate extracts against S. aureus, B. sub-tilis, and E. coli was evaluated using a well-diffusion method, and MIC was determined by the microdilution method. Cell lysis was observed through Transmission Electron Microscopy. Results: A BLAST search of 18S rRNA gene sequences of the marine isolates GSBT S13 and GSBT S14 showed 99.3 % sequence similarity with A. glaucus for both isolates and that of endophyte GSBT E3 showed 99.7 % sequence similarity with B. pinkertoniae. Cellulase pro-duction was comparatively higher in GSBT E3 and lipase from GSBT S13 and GSBT S14. Ethyl acetate extracts of GSBT S14 and GSBT E3 showed a clear zone of inhibition by the well-diffusion method, further confirmed by electron microscopy. HR-TEM showed that the ethyl acetate extracts of the isolates appeared to damage the cell membrane, leading to cell shrinkage and death in E. coli and S. aureus. Conclusion: GSBT S13 and GSBT S14 exhibited extracellular amylase, cellulase, and lipase activities. Ethyl acetate extracts of both GSBT S14 and B. pinkertoniae GSBT E3 showed better antibacterial properties against S. aureus.

Characterization of Pseudomonas spp. contamination and in situ spoilage potential in pasteurized milk production process

To investigate the prevalence of Pseudomonas in the pasteurized milk production process and its effect on milk quality, 106 strains of Pseudomonas were isolated from the pasteurized milk production process of a milk production plant in Shaanxi Province, China. The protease, lipase and biofilm-producing capacities of the 106 Pseudomonas strains were evaluated, and the spoilage enzyme activities of their metabolites were assessed by simulating temperature incubation in the refrigerated (7 °C) and transport environment (25 °C) segments and thermal treatments of pasteurization (75 °C, 5 min) and ultra-high temperature sterilization (121 °C, 15 s). A phylogenetic tree was drawn based on 16S rDNA gene sequencing and the top 5 strains were selected as representative strains to identify their in situ spoilage potential by examining their growth potential and ability to hydrolyze proteins and lipids in milk using growth curves, pH, whiteness, Zeta-potential, lipid oxidation, SDS-PAGE and volatile flavor compounds. The results showed that half and more of the isolated Pseudomonas had spoilage enzyme production and biofilm capacity, and the spoilage enzyme activity of metabolites was affected by the culture temperature and sterilization method, but ultra-high temperature sterilization could not completely eliminate the enzyme activity. The growth of Pseudomonas lundensis and Pseudomonas qingdaonensis was less affected by temperature and time, and the hydrolytic capacity of extracellular protease and lipase secreted by Pseudomonas lurida was the strongest, which had the greatest effect on milk quality. Therefore, it is crucial to identify the key contamination links of Pseudomonas, the main bacteria responsible for milk spoilage, and the influence of environmental factors on its deterioration.

Effect of m-PEG modification on the properties and structure of recombinant lipase from Aspergillus niger GZUF36

Natural lipase is challenging to meet industrial demands because of its low activity, poor stability, and intolerance to organic solvents. Our research aimed to improve the catalytic performance of the extracellular lipase from Aspergillus niger GZUF36 (PEXANL1) by modifying it with a molecular weight of 5000 monomethoxy polyethylene glycol (m-PEG5000) activated by p-toluenesulfonyl chloride (m-PEG5000-OTe). After modification of PEXANL1 with m-PEG5000-OTe (m-PEG5000-OTe@PEXANL1), the activity of m-PEG5000-OTe@PEXANL1 increased by 2.38-fold, its stability at high temperatures improved, and the optimal pH shifted towards the neutral region (pH 6). The effect of surfactants Triton X-100, Tween 20, Tween 80, and SDS on the enzyme activity of m-PEG5000-OTe@PEXANL1 was 118.44%, 124.28%, 120.40%, and 108.53%, respectively. They all activated the enzyme activity. Additionally, m-PEG5000-OTe@PEXANL enhanced its tolerance in organic solvents such as xylene and isopropanol. We analyzed the electrostatic interactions using Zeta potential and characterized the secondary structure using circular dichroism (CD) spectroscopy. The results indicated that the chemical modification led to structural changes in PEXANL1, enhancing its performance. The m-PEG5000-OTe@PEXANL1 may be used to synthesize healthy lipid 1, 3-diglyceride. That study laid the foundation for further industrial applications of PEXANL1 or other lipase by improving the catalytic performance of lipase through chemical modification.

Phenotypic and genetic characterization of a lipolytic Ralstonia mannitolilytica isolate from petroleum-contaminated soil in Iraq.

Lipases play a crucial role in various industrial applications, and microbial lipases, particularly those from bacteria, possess significant properties. With increasing concerns about the environmental and health impacts of hydrocarbons from pipelines and refineries, there is a growing need to mitigate the risks associated with these compounds. In this study, 40 bacterial isolates were recovered from contaminated soil samples collected from multiple refineries across Iraq. Using the Vitek system, bacterial isolates were identified up to the species level, revealing that only 12 isolates exhibited lipase-producing capabilities. Among the lipase-producing isolates, Ralstonia mannitolilytica demonstrated the highest extracellular lipase activity, as determined by an olive oil plate assay supplemented with rhodamine B. Confirmation of the species identity was achieved through 16S rRNA gene sequencing, with the obtained sequence deposited under accession number LC772176.1. Further sequence analysis revealed single nucleotide polymorphisms (SNPs) in the genome of Ralstonia mannitolilytica strain H230303-10_N19_7x_R2 (CP011257.1, positions 1,311,102 and 1,311,457). Additionally, the presence of the lipase gene was confirmed through amplification and sequencing using a thermocycler PCR. Sequence analysis of the gene, aligned using Geneious Prime software, identified SNPs (CP010799, CP049132, AY364601, CP011257, and CP023537), and a phylogenetic tree was constructed based on genetic characterization. Our findings highlight the potential of Ralstonia mannitolilytica as a promising candidate for lipase production and contribute to our understanding of its genetic diversity and biotechnological applications in hydrocarbon degradation and industrial processes.

Burkholderia cepacia: Understanding Pathogenicity, Virulence Factors, and Therapeutic Strategies

Burkholderia cepacia, formerly described as Pseudomonas cepacia, belongs to the Burkholderia cepacia complex (Bcc), a collection of closely related gram-negative bacteria that is considered a leading organism in causing pulmonary infections in humans. In healthcare settings, Burkholderia species have the ability to colonise medical equipment and surfaces, posing a threat of nosocomial infections. Moreover, they exhibit inherently resistance to various classes of antibiotics, thus being nominated as multi-drug resistant pathogens (MDR), that is why treatment of infections caused by these organisms is challenging. Enzymes such as metalloproteases, serine proteases, metalloproteases, and other extracellular lipases play a major role in its pathogenesis. In addition to this, different components, such as pili, flagella, and lipopolysaccharide (LPS), have a role in both the motility of the bacterial cell as well as its adherence to host cells. Identification is usually done with different biochemical reactions such as late oxidase activity, oxidation of glucose, variable lysine decarboxylation, maltose, mannitol, and lactose, and hydrogen sulphide gas production. Advanced techniques like semi-nested PCR, real-time PCR, or MALDI-TOF mass spectrometry enable quick and precise identification.&#x0D; Key words: Multi-drug resistant, Polymerase chain reaction, Burkholderia cepacia complex, matrix assisted laser desorption ionisation, time of flight mass spectrometry

Molecular Cloning of the Extracellular Lipases of Bacillus Amyloliquefaciens Isolated from Agrifood Wastes.

The lipase enzyme (EC: 3.1.1.3) is one of the most important catalysts in food, dairy, detergent, and textile industries. This study was performed to identify, isolate and characterize of lipase producing bacterial strain from agrifood wastes and to identify and characterize of their lipase genes. In the present study, two lipase-producing isolates were identified from the effluent of Golbahar meat products and Soveyda vegetable oil factories using in silico and in vitro approaches. The results of morphological, biochemical, and molecular characterizations showed that both lipase-producing isolates belong to the Bacillus amyloliquefaciens species. Phylogenetic analysis confirmed the results of phenotypic, biochemical, and molecular characterizations. The results showed differences between LipA and LipB in the Golbahar and Soveyda isolates. Three different amino acids (residues 14, 100, and 165) were observed in LipA and one different amino acid (residue 102) was detected in LipB extracellular lipases. The proteinmolecular weightof the two extracted lipases ranged from 20 to 25 kDa. The identified extracellular lipases also had different physicochemical features. The maximum lipase activity of the Golbahar and Soveyda isolates was observed at 45 °C and at the pH of 8, but the Golbahar isolates exhibited higher lipase activity compared to the Soveyda isolates. The Golbahar and Soveyda isolates exhibited different activities in the presence of some ions, inhibitors, denaturing agents, and organic solvents and the Golbahar isolates showed better lipase activity than the Soveyda isolates. In this study, two extracellular lipase-producing isolates of B. amyloliquefaciens were identified from different food industries, and their characteristics were investigated. The results of various investigations showed that the lipases produced by the Golbahar isolate have better characteristics than the lipases of the Soveyda isolate. The Golbahar lipases have a suitable temperature and pH activity range and maintain their activity in the presence of some ions, inhibitors, denaturing agents, and organic solvents. After further investigation, the Golbahar isolate lipase can be used in various industries. In addition, this lipase can be used enzyme engineering processes and its activity can be arbitrarily changed by targeted mutations. The results of this study can increase our knowledge of extracellular lipases and may turn out to have industrial applications.

New Bioremediation by Lipase Purified from Ralstonia mannitolilytica for Petroleum Hydrocarbons

A growing issue of contaminated soil caused by oil derivers in Iraq, requires detection of new methods for bioremediation. The present work aims to purify of extracellular lipase enzyme that can eliminate oil derivers. Seven strains of lipolytic bacteria were collected from Al- Dora electricity station soil Baghdad/ Iraq that polluted with oil derivers. Ralstonia mannitolilytica considered higher ability in lipolytic enzymes production. Crude lipase activity gave (1.66 U/ml). The purification procedure resulted in 8.9 fold with a 23.2% final yield. The purity of lipase and molecular weight confirmed by SDS-PAGE. The efficiency of ten aliphatic hydrocarbon compounds in the soil contaminated with diesel decreases after treatment with purified enzyme (7.6 U/ml) for 24 hr. at natural environment to mimic the nature compared with control. The results concluded the degradation of aliphatic hydrocarbon compounds by lipase enzyme that purified from local Ralstonia mannitolilytica isolates was an efficient (biological) treatment. Although there are studies related to bioremediation used different bacterial isolates types and enzymes such as Pseudomonas sp. none of it used Ralstonia mannitolilytica isolate and lipase enzyme as bioremediation of oil spills.

Effects of Phomopsidione on the Viability, Virulence, and Metabolites Profile of Methicillin-Resistant Staphylococcus aureus (MRSA).

Methicillin-resistant Staphylococcus aureus (MRSA) infections have become one of the most threatening multidrug-resistant pathogens. Thus, an ongoing search for anti-MRSA compounds remains an urgent need to effectively treating MRSA infections. Phomopsidione, a novel antibiotic isolated from Diaporthe fraxini, has previously demonstrated potent anti-candidal activity. The present study aimed to investigate the effects of phomopsidione on the viability, virulence, and metabolites profile of MRSA. MRSA was sensitive to phomopsidione in a concentration-dependent manner. Phomopsidione exhibited minimum inhibitory concentration and minimum bactericidal concentration of 62.5 and 500.00µg/mL against MRSA on broth microdilution assay. The compound showed significant reduction in virulence factors production including extracellular polymeric substances quantification, catalase, and lipase. An untargeted metabolomics analysis using liquid chromatography-high resolution mass spectrometry revealed a significant difference in the metabolites profile of MRSA with 13 putatively identified discriminant metabolites. The present study suggested the potential of phomopsidione as a promising anti-MRSA agent.

Waste cooking oil and molasses for the sustainable production of extracellular lipase by Saitozyma flava.

Organic waste valorization is one of the principal goals of the circular economy. Bioprocesses offer a promising approach to achieve this goal by employing microorganisms to convert organic feedstocks into high value products through their metabolic activities. In this study, a fermentation process for yeast cultivation and extracellular lipase production was developed by utilizing food waste. Lipases are versatile enzymes that can be applied in a wide range of industrial fields, from detergent, leather, and biodiesel production to food and beverage manufacturing. Among several oleaginous yeast species screened, Saitozyma flava was found to exhibit the highest secreted lipase activity on pNP-butyrate, pNP-caproate, and pNP-caprylate. The production medium was composed of molasses, a by-product of the sugar industry, which provided nutrients for yeast biomass formation. At the same time, waste cooking oil was employed to induce and enhance extracellular lipase production. After 48h of process, 20g/L of yeast biomass and 150mU/mgdw of lipase activity were achieved, with a productivity of 3mU/mgdw/h. The purified lipase from S. flava showed optimal performances at temperature 28°C and pH 8.0, exhibiting a specific activity of 62U/mg when using p-NPC as substrate.

Characterization of a recombinant Aspergillus niger GZUF36 lipase immobilized by ionic liquid modification strategy

Enzyme immobilized on magnetic nanomaterials is a promising biocatalyst with efficient recovery under applied magnets. In this study, a recombinant extracellular lipase from Aspergillus niger GZUF36 (PEXANL1) expressed in Pichia pastoris GS115 was immobilized on ionic liquid-modified magnetic nano ferric oxide (Fe3O4@SiO2@ILs) via electrostatic and hydrophobic interaction. The morphology, structure, and properties of Fe3O4@SiO2@ILs and immobilized PEXANL1 were characterized by scanning electron microscopy, Fourier transform infrared spectroscopy, x-ray diffraction, vibration sample magnetometer, and zeta potential analysis. Under optimized conditions, the immobilization efficiency and activity recovery of immobilized PEXANL1 were 52 ± 2% and 122 ± 2%, respectively. The enzymatic properties of immobilized PEXANL1 were also investigated. The results showed that immobilized PEXANL1 achieved the maximum activity at pH 5.0 and 45 °C, and the lipolytic activity of immobilized PEXANL1 was more than twice that of PEXANL1. Compared to PEXANL1, immobilized PEXANL1 exhibited enhanced tolerance to temperature, metal ions, surfactants, and organic solvents. The operation stability experiments revealed that immobilized PEXANL1 maintained 86 ± 3% of its activity after 6 reaction cycles. The enhanced catalytic performance in enzyme immobilization on Fe3O4@SiO2@ILs made nanobiocatalysts a compelling choice for bio-industrial applications. Furthermore, Fe3O4@SiO2@ILs could also benefit various industrial enzymes and their practical uses.Key points• Immobilized PEXANL1 was confirmed by SEM, FT-IR, and XRD.• The specific activity of immobilized PEXANL1 was more than twice that of PEXANL1.• Immobilized PEXANL1 had improved properties with good operational stability.Graphical abstract