Extracellular Lipase Research Articles

Identification of a lipolytic <em>Trichoderma</em> sp. and characterization of its extracellular lipase

The Ceylon Journal of Science is a peer-reviewed journal published quarterly by the University of Peradeniya, Sri Lanka in March, June, September and December. It is aimed at publishing high quality research articles on topics related to different disciplines in Science. The journal accepts original research articles, book reviews, reviews and mini-reviews, short communications, opinions, research notes, and commentaries and notes. The Journal has its own website https://site.pdn.ac.lk/cjs/. The journal strictly adheres to publication ethics as emphasized by the Committee on Publication Ethics (COPE). It is indexed in Sri Lanka Journals Online (SLJOL), Directory of Open Access Journals (DOAJ), Google Scholar and Zoological Records. According to the Google Scholar;H5-Index: 12H5-Median: 15According to the Exaly (1970 – 2021);Impact Factor: 0.6 (top 19%)Extended IF: 0.6 (top 19%)H-Index: 8 (top 28%)Citations/paper: 1.42

Study of Hydrolytic Enzymes Activity and Stability of the Isolated Yeast Close to Zygoascus hellenicus

Background: Lipase is one of the most well-known and essential biocatalysts in the de-tergent, food, and pharmaceutical industries. Microbial lipase sources such as yeasts are applicable due to their stability in harsh conditions. Objective: In this study, the effect of temperature, initial pH, and incubation time were investigated to improve the extracellular lipase production by yeast, named Zygoascus hellenicus strain MZ_574439 T. Methods: Strain MZ_574439 T has already been isolated and registered from Iran. In the current project, strain MZ_574439 T with 99% similarity to Z. hellenicus was isolated from water samples. Results: Our findings showed that the isolated strain has a remarkable difference from its close phy-logenetic species in the production of lipase and can produce extracellular lipase up to 7.2 U/ml while the Z. hellenicus has no ability of lipase production. The isolated strain was not able to pro-duce other hydrolytic enzymes. The enzyme activity results showed that the best activity for the iso-lated lipase is pH= 7 and 37°C. The best stability condition for the enzyme occurs at 50°C and pH =7. Conclusion: From the current study, it can be concluded that Z. hellenicus produces lipase. The li-pase enzyme production was optimized with different physiological conditions. Yeast extract could be a better source for maximum lipase production.

Role of cuticle-degrading enzymes of Beauveria bassiana and Metarhizium anisopliae in virulence on Plodia interpunctella (Lepidoptera, Pyralidae) larvae

Entomopathogenic fungi (EPF) are important biological control agents in pest management programs in agroecosystems against insect pests. EPF such as Beauveria bassiana (Bals.) Vuillemin and Metarhizium anisopliae (Metchn.) Sorokin produce a wide range of extracellular enzymes involved in disturbance of the first barrier in the insect cuticle comprising proteins, chitin, and lipids. Realizing relationships between the expression of these enzymes and fungal virulence might aid in development of effective mycoinsecticides. The virulence of B. bassiana (isolates TV and OZ1) and M. anisopliae (isolate CS1) were investigated on Plodia interpunctella (Hübner) larvae in this study. The third instar larvae were immersed in a suspension containing 1 × 108 conidia.mL-1 of fungal conidial inoculum. The results revealed that all three fungal isolates caused mortality in larvae, but there was a considerable variation in their virulence. Total proteinase, exochitinase and lipase assays were done for these isolates. The TV isolate with the highest mortality with 41.7%, had the highest level of specific activity of exochitinase, protease, and lipase with 0.148, 0.654, and 0.190 U. mg-1 protein, respectively. In the current study, a positive correlation was determined between the virulence of fungal isolates and the activities of protease and lipase, but this link was not significant for exochitinase. Our results demonstrated that extracellular enzymes, particularly protease and lipase, may play a crucial role in the virulence of these fungal isolates against P. interpunctella larvae.

A Study on the Isolation and Characterization of Bacillus subtilis Strains with Extracellular Lipase Production from Specific Environments

A Study on the Isolation and Characterization of Bacillus subtilis Strains with Extracellular Lipase Production from Specific Environments

Identification by Reverse Vaccinology of Three Virulence Factors in Burkholderia cenocepacia That May Represent Ideal Vaccine Antigens.

The Burkholderia cepacia complex comprises environmental and clinical Gram-negative bacteria that infect particularly debilitated people, such as those with cystic fibrosis. Their high level of antibiotic resistance makes empirical treatments often ineffective, increasing the risk of worst outcomes and the diffusion of multi-drug resistance. However, the discovery of new antibiotics is not trivial, so an alternative can be the use of vaccination. Here, the reverse vaccinology approach has been used to identify antigen candidates, obtaining a short-list of 24 proteins. The localization and different aspects of virulence were investigated for three of them-BCAL1524, BCAM0949, and BCAS0335. The three antigens were localized in the outer membrane vesicles confirming that they are surface exposed. We showed that BCAL1524, a collagen-like protein, promotes bacteria auto-aggregation and plays an important role in virulence, in the Galleria mellonella model. BCAM0949, an extracellular lipase, mediates piperacillin resistance, biofilm formation in Luria Bertani and artificial sputum medium, rhamnolipid production, and swimming motility; its predicted lipolytic activity was also experimentally confirmed. BCAS0335, a trimeric adhesin, promotes minocycline resistance, biofilm organization in LB, and virulence in G. mellonella. Their important role in virulence necessitates further investigations to shed light on the usefulness of these proteins as antigen candidates.

Serratia marcescens Necrotizing Fasciitis of the Ankle: A Case Report.

Necrotizing fasciitis is a devastating inflammatory infection requiring emergent medical treatment and surgical intervention. Even with timely management, the mortality rate of necrotizing fasciitis approaches 25%. The causative bacteria invade fascial planes and express toxins that advance rapidly. Here, we document a rare case of necrotizing fasciitis from Serratia marcescens infection. Serratia marcescens is capable of inducing a necrotizing inflammatory cascade mediated by extracellular cytotoxin and lipase. In this case report, a 90-year-old man presented to our emergency department from a long-term care facility with a relatively benign-appearing ulcer with surrounding cellulitis on the right ankle. Blood cultures and wound cultures confirmed the organism to be S marcescens. A multidisciplinary team was consulted for management. The patient received antibiotic therapy and medical support, but because of his comorbid conditions and social situation, the designated medical decision maker opted for comfort care rather than aggressive surgical debridement. The patient progressed through the clinical stages of necrotizing fasciitis. Within 36 hours, the patient died as result of sepsis-induced organ failure.

Genetic and Transcriptional Regulatory Mechanisms of Lipase Activity in the Plant Pathogenic Fungus Fusarium graminearum.

Lipases, which catalyze the hydrolysis of long-chain triglycerides, diglycerides, and monoglycerides into free fatty acids and glycerol, participate in various biological pathways in fungi. In this study, we examined the biological functions and regulatory mechanisms of fungal lipases via two approaches. First, we performed a systemic functional characterization of 86 putative lipase-encoding genes in the plant-pathogenic fungus Fusarium graminearum. The phenotypes were assayed for vegetative growth, asexual and sexual reproduction, stress responses, pathogenicity, mycotoxin production, and lipase activity. Most mutants were normal in the assessed phenotypes, implying overlapping roles for lipases in F. graminearum. In particular, FgLip1 and Fgl1 were revealed as core extracellular lipases in F. graminearum. Second, we examined the lipase activity of previously constructed transcription factor (TF) mutants of F. graminearum and identified three TFs and one histone acetyltransferase that significantly affect lipase activity. The relative transcript levels of FgLIP1 and FGL1 were markedly reduced or enhanced in these TF mutants. Among them, Gzzc258 was identified as a key lipase regulator that is also involved in the induction of lipase activity during sexual reproduction. To our knowledge, this study is the first comprehensive functional analysis of fungal lipases and provides significant insights into the genetic and regulatory mechanisms underlying lipases in fungi. IMPORTANCE Fusarium graminearum is an economically important plant-pathogenic fungus that causes Fusarium head blight (FHB) on wheat and barley. Here, we constructed a gene knockout mutant library of 86 putative lipase-encoding genes and established a comprehensive phenotypic database of the mutants. Among them, we found that FgLip1 and Fgl1 act as core extracellular lipases in this pathogen. Moreover, several putative transcription factors (TFs) that regulate the lipase activities in F. graminearum were identified. The disruption mutants of F. graminearum-lipase regulatory TFs all showed defects in sexual reproduction, which implies a strong relationship between sexual development and lipase activity in this fungus. These findings provide valuable insights into the genetic mechanisms regulating lipase activity as well as its importance to the developmental stages of this plant-pathogenic fungus.

Optimization of fermentation medium components by response surface methodology (RSM) and artificial neural network hybrid with genetic algorithm (ANN-GA) for lipase production by Burkholderia cenocepacia ST8 using used automotive engine oil as substrate

Bioconversion of used automotive engine oil (UEO) into lipase was conducted via submerged fermentation by Burkholderia cenocepacia ST8, as a strategy for value-added product generation and waste management. Response surface methodology (RSM) and artificial neural network hybrid with genetic algorithm (ANN-GA) were employed to optimize the fermentation medium for enhancing extracellular lipase production by B. cenocepacia ST8. Employing a four-factor-five-level central composite rotatable experimental design (CCRD), a reduced quartic polynomial RSM model and ANN model (4-4-1) trained using Bayesian Regularization were developed to attain the optimized fermentation medium for maximum level of lipase production. The RSM model predicted the following as the optimum media constituents: 2.28% v/v of Tween 80, 2.26% v/v of UEO, 0.79% w/v of nutrient broth, and 1.33% w/v of gum arabic, with an actual lipase yield of 216 U/mL. While, ANN-GA predicted the optimum media constituents to be 3% v/v of Tween 80, 3% v/v of UEO, 0.72% w/v of nutrient broth, and 3.38% w/v of gum arabic, with actual lipase yield of 225 U/mL. In comparison to the unoptimized medium, optimized RSM and ANN-GA systems both demonstrated a 1.6-fold increment in lipase production. Tween 80 and nutrient broth concentrations were the most important variables influencing the lipase production. The findings of this study indicated that the ANN-GA and RSM could be useful for effective optimization of the fermentation medium for enzyme production.

Scale-Up Lipase Production and Development of Methanol Tolerant Whole-Cell Biocatalyst from Magnusiomyces spicifer SPB2 in Stirred-Tank Bioreactor and Its Application for Biodiesel Production

This study aimed to economically develop the yeast whole-cell biocatalyst from the lipase-secreting Magnusiomyces spicifer SPB2 to serve green biodiesel production. The scaled-up productions of lipases were optimized using a 5-L stirred-tank bioreactor. The maximum extracellular lipase and cell-bound lipase (CBL) yields of 1189.65 U/L and 5603.74 U/L were achieved at 24 h and 60 h, respectively, in the modified IMY medium (pH 5.0) containing 2% of soybean oil as a carbon source and 0.2% Gum Arabic as an emulsifying agent. The optimized cultivation was initiated with an inoculum size of 1 × 107 cells/mL and conducted under an aeration rate of 0.75 vvm with an agitation speed of 400 rpm. The obtained whole-cell biocatalyst of M. spicifer SPB2 was applied to catalyze the transesterification reaction using palm oil and methanol as substrates. The greatest yield of 97.93% fatty acid methyl ester (FAME) was reached at 72 h using a palm oil/methanol ratio of 1:7, indicating high methanol stability of the biocatalyst. Moreover, substrate homogenization accelerated the reaction to achieve FAME production of 97.01% at 48 h and remained stable afterwards. Without homogenization, the highest FAME of 98.20% was obtained at 60 h. The whole-cell biocatalyst prepared from lipase-secreting M. spicifer SPB2 at an up-scaled level greatly enhanced efficiency and feasibility for commercial biodiesel production through a green conversion process.

Production of Methanol Tolerant Cell-bound Lipase of Magnusiomyces spicifer SPB2 and Application as Whole-cell Biocatalyst in Transesterification Reaction

Cell-bound lipase (CBL) is a potential biocatalyst for production of fatty acid alkyl ester (FAAE), which is feasible replacement for diesel fuel. This study aimed to improve both hydrolysis and transesterification activities of CBL from the yeast Magnusiomyces spicifer SPB2, which could be employed as whole-cell biocatalyst for transesterification reactions through optimizing the growth medium using Taguchi orthogonal array experimental method (L9). The optimized medium, resulting the highest CBL transesterification activity, consisted of soybean oil, peptone, Gum Arabic and an initial pH of 5. Interestingly, the fatty acid methyl ester (FAME) yield increased up to 33.28 times when compared with non-optimized medium. Moreover, cell morphology of M. spicifer SPB2 varied depending on the culture medium component. The single cell was more active to be used as the whole-cell biocatalyst than the pseudomycelial form for transesterification reaction. The highest FAME yield was 93.86 % when refined palm oil to methanol molar ratio of 1:8 was used as substrates. Whole-cell lipase from M. spicifer showed favorable activity to iso-butanol as an acyl accepter, resulting in fatty acid butyl ester (FABE) yield of 86.80 % at 1:3 molar ratio of palm oil to iso-butanol. The FAME and FABE yields dramatically decreased from 93.86 and 86.80 % to 57.02 and 4.48 %, respectively after the reuse of the enzyme to the second batch of reaction. These results indicated that the whole-cell biocatalyst developed from M. spicifer was an efficient and economical approach to improve enzyme activity and stability in the transesterification reaction. HIGHLIGHTS Methanol tolerant cell-bound lipase and extracellular lipase of spicifer SPB2 were produced The whole-cell biocatalyst of spicifer SPB2 was developed for FAME production The optimized medium was developed to prepare whole-cell lipases from spicifer SPB2 Gum Arabic was the most influencing factor for synthetic activity of whole-cell CBL from yeast Single cell morphology exhibited higher transesterification activity than pseudomycelial form GRAPHICAL ABSTRACT

Purification and characterization of lipase produced from Bacillus cereus (PCSIR NL-37)

There is a growing trend to produce lipase from microorganisms owing to their commercial demand in various industries. Bacillus cereus has been shown to have extracellular lipase activity and high growth rates. This study explains the purification of microbial lipase to homogeneity by dialysis, precipitation and chromatography. The purified enzyme with 56kDa relative molecular mass exhibited the highest activity at 60°C (95.56U/ml) and pH 7 (124.50U/ml). The enzyme activity was highly promoted in the presence of K+ (136.17U/ml) and Zn++(133.07 U/ml), and SDS did not affect the enzyme activity, whereas in the company of triton X100 activity of lipase is maximum (23.90 U/ml). The enzyme activity was enhanced by using almond oil (120.00 U/ml) as a substrate. We deduce cheaper protocols for producing extracellular lipase via simple laboratory techniques, which could be a good insight for its production at the commercial level. Keywords: Lipase; Bacillus cereus; chromatography; enzyme activity; purification.

Screening, identification, and characterization of lipase-producing halotolerant Bacillus altitudinis Ant19 from Antarctic soil.

Potent lipase-producing and halotolerant Bacillus altitudinis Ant19 strain was screened and isolated from Antarctic soil. The isolate showed broad-range lipase activity against different lipid substrates. Presence of lipase activity was confirmed by PCR amplification and sequencing of the lipase gene from Ant19. The study attempted to establish the use of crude extracellular lipase extract as cheap alternative to purified enzyme by characterizing the crude lipase activity and testing it in certain practical applications. Crude lipase extract from Ant19 showed high stability at 5-28℃ (> 97%), while lipase activity was noted in a wide temperature range of 20-60℃ (> 69%), with optimum activity at 40℃ (117.6%). The optimum lipolytic activity was noted at pH 8 with good activity and stability in alkaline conditions (pH 7-10). Moreover, the lipase activity was substantially stable in various solvents, commercial detergents, and surfactants. It retained 97.4% activity in 1% solution of commercial Nirma detergent. Besides, it was non-regiospecific, and active against substrates having different fatty acid chain lengths with preference for shorter chain length. Further, the crude lipase enhanced the oil stain removal efficiency of commercial detergent from 52 to 77.9%, while 66% oil stain was removed using crude lipase alone. Immobilization process improved the storage stability of crude lipase for 90days. In our knowledge, it is the first study on characterization of lipase activity from B. altitudinis, which has promising applications in various fields.

Homologous High-Level Lipase and Single-Cell Protein Production with Engineered Yarrowia lipolytica via Scale-Up Fermentation for Industrial Applications

Yarrowia lipolytica is a promising feed additives. Here, we aimed to produce extracellular lipases and single-cell proteins (SCPs) at high levels simultaneously through fed-batch fermentation of engineered Y. lipolytica. The parameters for 500 mL shake flask cultures were optimized with a single factorial design. The resultant activity of lipase reached 880.6 U/mL after 84 h of fermentation, and 32.0 g/L fermentation broth of dry SCP was obtained at 120 h. To attain high SCP and lipase productivity, the high-density fed-batch fermentation of Y. lipolytica was scaled up in 10 L, 30 L, and 100 L fermentors. Using glycerol as the sole carbon source, the lipase activity peaked to 8083.3 U/mL, and the final dry SCP weight was 183.1 g/L at 94.6 h in 10 L fermentors. The extracellular lipase activity and SCP weight reached 11,100.0 U/mL and 173.3 g of dry SCP/L at 136 h in 30 L fermentors, respectively. Following 136 h of fed-batch fermentation, the extracellular lipase activity and dry SCP weight reached 8532.0 U/mL and 170.3 g/L in 100 L fermentors, respectively. A balance between the lipase secretion and growth of Y. lipolytica recombinant strain was achieved, indicating that an efficient fermentation strategy could promote further scale-up for industrial SCP production from engineered Y. lipolytica.

Physiological and Biochemical Characteristics, and Biotechnological Potential of Hydrolitic Haloalkalitolerant Bacteria of Soda Sludge Storage

Peculiarities of manifestation of hydrolytic activity of haloalkalitolerant bacteria Pseudomonas peli 3-T, Microbacterium kitamiense 16-DB, and Bacillus aequororis 5-DB, isolated from the soil of the territory of the soda sludge storage and the soda sludge of the functioning sludge reservoir of Berezniki Soda Plant (Perm Territory, Russia) were studied. It was shown that the maximum activity of extracellular lipases and amylases of the three studied bacteria were manifested at pH 11 and a high concentration of sodium chloride (50–200 g/L). Cultivation of P. peli 3-T on a mineral medium with 0.5% glycerol (source of carbon) and 0.03% urea (source of nitrogen) makes it possible to obtain biomass (6.9 g/L) and activity of extracellular lipase (1.26 U/L) and lipase associated with cells (3.02 U/mg dry cells) with a high economic ratio of substrate consumption (138%). The efficiency of immobilization of P. peli 3-T and B. aequororis 5-DB cells for use in lipid and starch biotransformation was shown. P. peli 3-T cells immobilized by adsorption on kaolin and incorporated into the structure of barium alginate and agarose gels retained from 40.4 to 63.8% of the cell lipase activity in suspension. B. aequororis 5-DB cells immobilized by adsorption on kaolin retained 42.5% of lipase and 90.7% of amylase activity as compared to the initial ones. At the same time, the activities of the P. peli 3-T and B. aequororis 5-DB enzymes were preserved during 6 consecutive reactions. The amylase activity of immobilized M. kitamiense 16-DB cells by adsorption on kaolin and chitosan decreased to 2.7–3.5% of the initial value and was completely inhibited upon immobilization by cell’s entrapment into barium alginate gels or agarose gels.

Polymer-Degrading Enzymes of Pseudomonas chloroaphis PA23 Display Broad Substrate Preferences.

Although many bacterial lipases and PHA depolymerases have been identified, cloned, and characterized, there is very little information on the potential application of lipases and PHA depolymerases, especially intracellular enzymes, for the degradation of polyester polymers/plastics. We identified genes encoding an intracellular lipase (LIP3), an extracellular lipase (LIP4), and an intracellular PHA depolymerase (PhaZ) in the genome of the bacterium Pseudomonas chlororaphis PA23. We cloned these genes into Escherichia coli and then expressed, purified, and characterized the biochemistry and substrate preferences of the enzymes they encode. Our data suggest that the LIP3, LIP4, and PhaZ enzymes differ significantly in their biochemical and biophysical properties, structural-folding characteristics, and the absence or presence of a lid domain. Despite their different properties, the enzymes exhibited broad substrate specificity and were able to hydrolyze both short- and medium-chain length polyhydroxyalkanoates (PHAs), para-nitrophenyl (pNP) alkanoates, and polylactic acid (PLA). Gel Permeation Chromatography (GPC) analyses of the polymers treated with LIP3, LIP4, and PhaZ revealed significant degradation of both the biodegradable as well as the synthetic polymers poly(ε-caprolactone) (PCL) and polyethylene succinate (PES).

Lipases from different yeast strains: Production and application for n-3 fatty acid enrichment of tuna eyeball oil

Enrichment of n-3 polyunsaturated fatty acids (PUFA) in the glyceride fraction of skipjack tuna eyeballs oil (STEO) was investigated by using six extracellular and cell-bound lipases from three different yeast strains. Among all lipases tested, Candida rugosa extracellular lipase (Cr-ECL) showed promising enrichment of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in STEO. Firstly, various parameters including types of solvent, solvent to oil ratio, water to oil ratio, hydrolysis time, temperature, and lipase concentration were studied to select the major independent parameters. Finally, response surface methodology (RSM) was used to optimize the reaction conditions. Optimal condition included 450 U of Cr-ECL at water to oil ratio of 1:8 for 11.28 h, which resulted in 1.9-fold increase in DHA concentration in STEO. EPA was increased by 2.1-fold, when 191.35 U of Cr-ECL, water to oil ratio of 1:6 for 5.14 h was used. The Cr-ECL catalyzed hydrolysis was non-stereoselective, but had acyl chain specificity, and recognition of the whole triacyl-glycerol molecule as shown by 1H NMR and 13C NMR. Therefore, the established hydrolysis protocol for DHA and EPA enrichment was successful in converting fish oil from tuna waste to useful and healthy fish oil enriched with n-3 fatty acids. These findings pave the way for the future valorization of industrial marine waste oil.

Production of Enriched Biomass by Carotenogenic Yeasts Cultivated on by-Products of Poultry Processing—A Screening Study

Carotenogenic yeasts are a group of microorganisms producing valuable metabolites such as carotenoids, ergosterol, ubiquinone or fatty acids. Their exceptional adaptability allows them to grow in diverse conditions. Owing to their extracellular lipase activity, they are capable of processing many lipid-type waste substrates. This study discusses the processing of poultry waste, specifically fat and feathers by using carotenogenic yeasts. Poultry fat does not require any pre-treatment to be utilized by yeast, but hydrolytic pre-treatment is required for the utilization of the nitrogen contained in feathers. Glycerol was used as a supplementary substrate to support the culture in the early stages of growth. Seven yeast strains were used for the experiments, of which the strain Rhodotorula mucilaginosa CCY19-4-25 achieved exceptional results of biomass production: 29.5 g/L on poultry fat + 10% glycerol at C/N ratio 25 and 28.3 g/L on media containing poultry fat + 25% glycerol at C/N 50. The bioreactor cultivation of the Rhodosporidium toruloides strain in media containing glycerol and feather hydrolysate as a nitrogen substrate achieved a biomass yield of 34.92 g/L after 144 h of cultivation. The produced enriched yeast biomass can be used as a component for poultry feeding; thus, the study is performed under the biorefinery concept.

A comprehensive proteomics analysis of the response of Pseudomonas aeruginosa to nanoceria cytotoxicity

The increased commercial use and spread of nanoceria raises concerns about the risks associated with its effects on living organisms. Although Pseudomonas aeruginosa may be ubiquitous in nature, it is largely found in locations closely linked with human activity. P. aeruginosa san ai was used as a model organism for a deeper understanding of the interaction between biomolecules of the bacteria with this intriguing nanomaterial. A comprehensive proteomics approach along with analysis of altered respiration and production of targeted/specific secondary metabolites was conducted to study the response of P. aeruginosa san ai to nanoceria. Quantitative proteomics found that proteins associated with redox homeostasis, biosynthesis of amino acids, and lipid catabolism were upregulated. Proteins from outer cellular structures were downregulated, including transporters responsible for peptides, sugars, amino acids and polyamines, and the crucial TolB protein of the Tol-Pal system, required for the structural formation of the outer membrane layer. In accordance with the altered redox homeostasis proteins, an increased amount of pyocyanin, a key redox shuttle, and the upregulation of the siderophore, pyoverdine, responsible for iron homeostasis, were found. Production of extracellular molecules, e.g. pyocyanin, pyoverdine, exopolysaccharides, lipase, and alkaline protease, was significantly increased in P. aeruginosa san ai exposed to nanoceria. Overall, nanoceria at sublethal concentrations induces profound metabolic changes in P. aeruginosa san ai and provokes increased secretion of extracellular virulence factors, revealing the powerful influence this nanomaterial has on the vital functions of the microorganism.

Bioprospecting for Novel Bacterial Sources of Hydrolytic Enzymes and Antimicrobials in the Romanian Littoral Zone of the Black Sea.

Marine microorganisms have evolved a large variety of metabolites and biochemical processes, providing great opportunities for biotechnologies. In the search for new hydrolytic enzymes and antimicrobial compounds with enhanced characteristics, the current study explored the diversity of cultured and uncultured marine bacteria in Black Sea water from two locations along the Romanian coastline. Microbial cell density in the investigated samples varied between 65 and 12.7 × 103 CFU·mL-1. The total bacterial community identified by Illumina sequencing of 16S rRNA gene comprised 185 genera belonging to 46 classes, mainly Gammaproteobacteria, Alphaproteobacteria, Flavobacteriia, and 24 phyla. The 66 bacterial strains isolated on seawater-based culture media belonged to 33 genera and showed variable growth temperatures, growth rates, and salt tolerance. A great fraction of these strains, including Pseudoalteromonas and Flavobacterium species, produced extracellular proteases, lipases, and carbohydrases, while two strains belonging to the genera Aquimarina and Streptomyces exhibited antimicrobial activity against human pathogenic bacteria. This study led to a broader view on the diversity of microbial communities in the Black Sea, and provided new marine strains with hydrolytic and antimicrobial capabilities that may be exploited in industrial and pharmaceutical applications.

Novel perspective on the spoilage metabolism of refrigerated sturgeon fillets: Nonspecific spoilage dominant organisms play an important role

Aeromonas sobria, a specific spoilage organism (SSO) in refrigerated (4 °C) sturgeon fillets, is considered as the main contributor for sturgeon spoilage. However, the spoilage metabolism of A. sobria is influenced by bacterial interactions, and the effect of nonspecific spoilage dominant organisms on the spoilage metabolism of A. sobria during storage is still unknown. In this study, the spoilage characteristics and metabolite changes of different inoculation groups under chilled storage were investigated through a co-cultivation model. The results of spoilage indicators indicated that the microbial interactions changed the spoilage characteristic of the fish model, although A. sobria played the most important role. Based on metabolome results, 549 differential metabolites were identified in two ion modes, which involved 9 chemical categories and 4 main metabolic pathways. The nonspecific spoilage dominant bacteria M. caseolyticus in the early stage of refrigeration affects the amino acid metabolism pathways of fish model by secreting extracellular proteases and peptidases. However, the nonspecific spoilage dominant bacteria A. albensis in the later period of refrigeration accelerate the lipid metabolism, and amino acid metabolism via secreting extracellular peptidases, lipase and esterase. This work may provide insights on controlling the spoilage of aquatic products by intervening degradation pathways.