Recent advances in production and applications of ectoine, a compatible solute of industrial relevance.

Extremophilic bacteria growing in saline ecosystems are potential producers of biotechnologically important products including compatible solutes. Ectoine/hydroxyectoine are two such solutes that protect cells and associated macromolecules from osmotic, heat, cold and UV stress without interfering with cellular functions. Since ectoine is a high value product, overviewing strategies for improving yields become relevant. Screening of natural isolates, use of inexpensive substrates and response surface methodology approaches have been used to improve bioprocess parameters. In addition, genome mining exercises can aid in identifying hitherto unreported microorganisms with a potential to produce ectoine that can be exploited in the future. Application wise, ectoine has various biotechnological (protein protectant, membrane modulator, DNA protectant, cryoprotective agent, wastewater treatment) and biomedical (dermatoprotectant and in overcoming respiratory and hypersensitivity diseases) uses. The review summarizes current updates on the potential of microorganisms in the production of this industrially relevant metabolite and its varied applications.

Comprehensive updates on the biological features and metabolic potential of the versatile extremophilic actinomycete Nocardiopsis dassonvillei.

Nocardiopsis dassonvillei prevails under harsh environmental conditions and the purpose of this review is to highlight its biological features and recent biotechnological applications. The organism prevails in salt-rich soils/marine systems and some strains endure extreme temperatures and pH. A few isolates are associated with marine organisms and others cause human diseases. Comparative genomic analysis indicates its versatility in producing biotechnologically relevant metabolites. Antimicrobial, cytotoxic, anticancer and growth promoting biomolecules are obtained from this organism. It also synthesizes biotechnologically important enzymes. Bioactive compounds and enzymes obtained from this actinomycete provide evidence regarding its metabolic competence and its potential economic value.

Use of marine microorganisms in designing anti-infective strategies for sustainable aquaculture production.

Aquaculture, a noteworthy food production sector, is confronted with disease occurrences. Treatment of aquaculture pathogens with antibiotics is often rendered ineffective due to biofilm formation and the development of resistant strains. Marine ecosystems encompass unusual microorganisms that produce novel bioactive compounds, including agents that could be used as alternatives to antibiotics. Moreover, biomass and/or biomolecules associated with these microorganisms could act as feed supplements to enhance the overall health of aquaculture species' and improve water quality parameters. The present review summarizes the contents of studies on such marine microorganisms with the potential to be developed as agents for tackling bacterial diseases in the aquaculture segment. Bioactive compounds produced by marine bacteria are known to inhibit biofilm-associated infections mediated by their bactericidal properties (produced by Bacillus, Vibrio, Photobacterium, and Pseudoalteromonas species), surfactant activity (obtained from different species of Bacillus and Staphylococcus lentus), anti-adhesive activity (derived from Bacillus sp. and Brevibacterium sp.), and quorum sensing inhibition. Several marine fungal isolates capable of producing antibacterial agents have also been effective in inhibiting aquaculture-associated pathogens. Another strategy followed by investigators to reduce the severity of infections is the use of bacterial, yeast, and microalgae biomass as feed supplements, probiotics, and immunostimulants. In some cases, marine microalgae have been employed as sustainable alternatives to fish oil and fish meal without compromising on nutritional quality. Their inclusion in aquaculture feed has enhanced growth, favored better survival of cultured species, and improved water quality parameters. Marine microorganisms (by providing effective bioactive compounds and being used as feed supplements) could enable aquaculture practices to be more sustainable in the future.

Whole-Genome Sequencing of the Tropical Marine Bacterium Nocardiopsis dassonvillei NCIM 5124, Containing the Ectoine Biosynthesis Gene Cluster ectABC.

The genome sequence (7,057,619 bp; GC content, 72.07%) of a tropical marine isolate, Nocardiopsis dassonvillei NCIM 5124, containing the biomedically and biotechnologically important gene cluster ectABC is reported here.

A comprehensive assessment of Yarrowia lipolytica and its interactions with metals: Current updates and future prospective.

The non-conventional yeast Yarrowia lipolytica has been popular as a model system for understanding biological processes such as dimorphism and lipid accumulation. The organism can efficiently utilize hydrophobic substrates (hydrocarbons and triglycerides) thereby rendering it relevant in bioremediation of oil polluted environments. The current review focuses on the interactions of this fungus with metal pollutants and its potential application in bioremediation of metal contaminated locales. This fungus is intrinsically equipped with a variety of physiological and biochemical features that enable it to tide over stress conditions induced by the presence of metals. Production of enzymes such as phosphatases, reductases and superoxide dismutases are worth a special mention. In the presence of metals, levels of inherently produced metal binding proteins (metallothioneins) and the pigment melanin are seen to be elevated. Morphological alterations with respect to biofilm formation and dimorphic transition from yeast to mycelial form are also induced by certain metals. The biomass of Y. lipolytica is inherently important as a biosorbent and cell surface modification, process optimization or whole cell immobilization techniques have aided in improving this capability. In the presence of metals such as mercury, cadmium, copper and uranium, the culture forms nanoparticulate deposits. In addition, on account of its intrinsic reductive ability, Y. lipolytica is being exploited for synthesizing nanoparticles of gold, silver, cadmium and selenium with applications as antimicrobial compounds, location agents for bioimaging and as feed supplements. This versatile organism thus has great potential in interacting with various metals and addressing problems related to their pollutant status.

Assessment of recombinant glutathione-S-transferase (HaGST-8) silica nano-conjugates for effective removal of pesticides.

Diverse glutathione-S-transferases (GSTs) are produced by insect pests including Helicoverpa armigera (HaGSTs) for detoxification of insecticides or xenobiotic compounds that they encounter. In an earlier study, the HaGST-8 gene was isolated from H. armigera larvae exposed to pesticide mixtures and the recombinant protein was expressed in the yeast Pichia pastoris. In this investigation, HaGST-8 was successfully immobilized on glutaraldehyde-activated APTES functionalized silica nanoparticles to obtain SiAPT-HaGST-8 nano-conjugates. Although enzyme activity associated with these conjugates was comparable to that of free HaGST-8, the specific activity of the former was found to be 1.25 times higher than the latter. In comparison with the free enzyme (that demonstrated a pH optimum of 9.0), for the nano-conjugates, the pH range was extended between pH 8.0 to 9.0. The optimum temperature for activity of both forms of the enzyme was found to be 30 °C. Stability of the enzyme was improved from 20 d for free HaGST-8 to 30 d for SiAPT-HaGST-8 nano-conjugates. Some loss in GST activity was detected after every reuse cycle of nano-conjugates and in all, 63% reduction was observed after three cycles. When 3 kinds of pesticides (namely, chlorpyrifos, dichlorvos and cypermethrin) were reacted with SiAPT-HaGST-8, more than 80% reduction in levels were observed. On the basis of the results obtained, the use of such silica nanoparticle-based systems for stable enzyme conjugation followed by effective removal of pesticides from aqueous media is envisaged.

Layer-by-Layer Assembled Nanostructured Lipid Carriers for CD-44 Receptor-Based Targeting in HIV-Infected Macrophages for Efficient HIV-1 Inhibition.

Macrophages act as a cellular reservoir in HIV infection. Elimination of HIV from macrophages has been an unfulfilled dream due to the failure of drugs to reach them. To address this, we developed CD44 receptor-targeted, novel hyaluronic acid (HA)-coated nanostructured lipid carriers (NLCs) of efavirenz via washless layer-by-layer (LbL) assembly of HA and polyallylamine hydrochloride (PAH). NLCs were subjected to TEM analysis, size and zeta potential, in vitro release and encapsulation efficiency studies. The uptake of NLCs in THP-1 cells was studied using fluorescence microscopy and flow cytometry. The anti-HIV efficacy was evaluated using p24 antigen inhibition assay. NLCs were found to be spherical in shape with anionic zeta potential (-23.66 ± 0.87 mV) and 241.83 ± 5.38 nm particle size. NLCs exhibited prolonged release of efavirenz during in vitro drug release studies. Flow cytometry revealed 1.73-fold higher uptake of HA-coated NLCs in THP-1 cells. Cytotoxicity studies showed no significant change in cell viability in presence of NLCs as compared with the control. HA-coated NLCs distributed throughout the cell including cytoplasm, plasma membrane and nucleus, as observed during fluorescence microscopy. HA-coated NLCs demonstrated consistent and significantly higher inhibition (81.26 ± 1.70%) of p24 antigen which was 2.08-fold higher than plain NLCs. The obtained results suggested preferential uptake of HA-coated NLCs via CD44-mediated uptake. The present finding demonstrates that HA-based CD44 receptor targeting in HIV infection is an attractive strategy for maximising the drug delivery to macrophages and achieve effective viral inhibition.

Transcriptome Response of the Tropical Marine Yeast Yarrowia lipolytica on Exposure to Uranium.

In our earlier investigation, we reported the consequences of uranium (U)-induced oxidative stress and cellular defense mechanisms alleviating uranium toxicity in the marine yeast Yarrowia lipolytica NCIM 3589. However, there is lack of information on stress response towards uranium toxicity at molecular level in this organism. To gain an insight on this, transcriptional response of Y. lipolytica after exposure to 50 µM uranium was investigated by RNA sequencing at the global level in this study. The de novo transcriptome analysis (in triplicates) revealed 56 differentially expressed genes with significant up-regulation and down-regulation of 33 and 23 transcripts, respectively, in U-exposed yeast cells as compared to the control, U-unexposed cells. Highly up-regulated genes under U-treated condition were identified to be primarily involved in transport, DNA damage repair and oxidative stress. The major reaction of Y. lipolytica to uranium exposure was the activation of oxidative stress response mechanisms to protect the important biomolecules of the cells. On the other hand, genes involved in cell wall and cell cycle regulation were significantly down-regulated. Overall, the transcriptional profiling by RNA sequencing to stress-inducing concentration of uranium sheds light on the various responses of Y. lipolytica for coping with uranium toxicity, providing a foundation for understanding the molecular interactions between uranium and this marine yeast.

Removal of uranium by immobilized biomass of a tropical marine yeast Yarrowia lipolytica.

A marine yeast, Yarrowia lipolytica isolated from an oil polluted sea water and shown earlier to sequester dissolved uranium (U) at pH 7.5, was utilized in the present study for developing an immobilized-cell process for U removal from aqueous solutions under batch and continuous flow through systems. In batch system, optimum biosorption conditions for U removal were assessed by investigating the effects of biomass dose, initial U concentration, contact time and pH of solution using Y. lipolytica cells immobilized in calcium alginate beads. Appreciable uranium-binding capabilities over a wide pH range (3-9) were observed with the alginate beads bearing yeast cells. Out of Langmuir and Freundlich models employed for describing the sorption equilibrium data under batch mode, uranyl adsorption followed Langmuir approach with satisfactory correlation coefficient higher than 0.9. Uranyl adsorption kinetics by Y. lipolytica entrapped in alginate beads was best described by the pseudo-second-order model. While the environmental scanning electron microscopy established the immobilization and the uniform distribution of Y. lipolytica cells in the alginate beads, the Energy Dispersive X-ray spectroscopy analysis confirmed the deposition of U in the beads following their exposure to uranyl solution. Fixed bed flow-through column comprising of Y. lipolytica biomass immobilized in polyacrylamide matrix displayed high efficacy for continuous removal of uranium at pH 7.5 up to five adsorption-desorption cycles. Adsorbed U by immobilized cells could be significantly desorbed using 0.1 N HCl. Overall, our results present the superior efficiency of immobilized Y. lipolytica biomass for U removal using batch and regenerative approaches.

Evaluation of silica nanoparticle mediated delivery of protease inhibitor in tomato plants and its effect on insect pest Helicoverpa armigera.

The inert and surface tunable nature of silica nanoparticles (SiNPs) makes them suitable for different applications. We have evaluated the potential of SiNPs for delivering proteins in tomato (Lycopersicon esculentum) plants. SiNPs of 20 and 100 nm (Si20 and Si100) were functionalized with (3-aminopropyl) triethoxysilane (APTES) to obtain Si20APT and Si100APT, respectively, that were non-toxic toward plants. The functionalized nanoparticles were taken up by plants through roots as well as leaf surfaces. They were seen to be localized near the vasculature, particularly around the xylem. Si20APT and Si100APT nanoparticles were conjugated with soybean trypsin inhibitor (STI) to yield Si20APT-STI and Si100APT-STI, respectively. Based on the trypsin inhibitory activity of loaded nanoparticles, optimum loading was obtained for 0.4 mg of STI per 0.8 mg of NPs. Si20APT nanoparticles retained higher contents of STI than Si100APT. Exposure of STI-conjugated nanoparticles to 25°C or pH 8.0 aided release of the inhibitor. The particle bound STI inhibited bovine trypsin by 80% and Helicoverpa armigera gut proteinase (HGP) activity by 50%. Second instar H. armigera larvae ingesting STI-loaded particles (incorporated in artificial diet or leaves) showed significant retardation in growth. In choice assays, Si20APT-STI applied leaf discs were strikingly avoided by insect larvae. On the basis of the results obtained in this investigation, we recommend the use of Si20 nanoparticles for developing plant delivery vehicles in the future.

Uptake and detoxification of diesel oil by a tropical soil Actinomycete Gordonia amicalis HS-11: Cellular responses and degradation perspectives.

A tropical soil Actinomycete, Gordonia amicalis HS-11, has been previously demonstrated to degrade unsaturated and saturated hydrocarbons (squalene and n-hexadecane, respectively) in an effective manner. In present study, G. amicalis HS-11 degraded 92.85 ± 3.42% of the provided diesel oil [1% (v/v)] after 16 days of aerobic incubation. The effect of different culture conditions such as carbon source, nitrogen source, pH, temperature, and aeration on degradation was studied. During degradation, this Actinomycete synthesized surface active compounds (SACs) in an extracellular manner that brought about a reduction in surface tension from 69 ± 2.1 to 30 ± 1.1 mN m-1 after 16 days. The morphology of cells grown on diesel was monitored by using a Field Emission Scanning Electron Microscope. Diesel-grown cells were longer and clumped with smooth surfaces, possibly due to the secretion of SACs. The interaction between the cells and diesel oil was studied by Confocal Laser Scanning Microscope. Some cells were adherent on small diesel droplets and others were present in the non-attached form thus confirming the emulsification ability of this organism. The fatty acid profiles of the organism grown on diesel oil for 48 h were different from those on Luria Bertani Broth. The genotoxicity and cytotoxicity of diesel oil before and after degradation were determined. Cytogenetic parameters such as mitotic index (MI); mitosis distribution and chromosomal aberration (type and frequency) were assessed. Oxidative stress was evaluated by measuring levels of catalase, superoxide dismutase and concentration of malondialdehyde. On the basis of these studies it was deduced that the degradation metabolites were relatively non-toxic.

Gold nanoparticles biosynthesized by Nocardiopsis dassonvillei NCIM 5124 enhance osteogenesis in gingival mesenchymal stem cells.

Gold nanoparticles are widely used for biomedical applications owing to their biocompatibility, ease of functionalization and relatively non-toxic nature. In recent years, biogenic nanoparticles have gained attention as an eco-friendly alternative for a variety of applications. In this report, we have synthesized and characterized gold nanoparticles (AuNPs) from an Actinomycete, Nocardiopsis dassonvillei NCIM 5124. The conditions for biosynthesis were optimized (100 mg/ml of cell biomass, 2.5 mM tetrachloroauric acid (HAuCl4) at 80 °C and incubation time of 25 min) and the nanoparticles were characterized by TEM, SAED, EDS and XRD analysis. The nanoparticles were spherical and ranged in size from 10 to 25 nm. Their interactions with human gingival tissue-derived mesenchymal stem cells (GMSCs) and their potential applications in regenerative medicine were evaluated further. The AuNPs did not display cytotoxicity towards GMSCs when assessed by 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide assay, DNA fragmentation patterns and Annexin V/propidium iodide staining techniques. These AuNPs induced faster cell migration when monitored by the in vitro wound healing assay. The effect of these nanoparticles on osteogenesis of GMSCs was also studied. Based on the results obtained from alkaline phosphatase, Von Kossa staining and Alizarin Red S staining, the AuNPs were seen to positively affect differentiation of GMSCs and enhance mineralization of the synthesized matrix. We therefore conclude that the biogenic, non-toxic AuNPs are of potential relevance for tissue regeneration applications.

Impact of uranium exposure on marine yeast, Yarrowia lipolytica: Insights into the yeast strategies to withstand uranium stress.

A marine yeast, Yarrowia lipolytica, was evaluated for morphological, physiological and biochemical responses towards uranium (U) exposure at pH 7.5. The yeast revealed biphasic U binding - a rapid biosorption resulting in ∼35% U binding within 15-30 min followed by a slow biomineralization process, binding up to ∼45.5% U by 24 h on exposure to 50 µM of uranyl carbonate. Scanning electron microscopy coupled with Energy Dispersive X-ray spectroscopy analysis of 24 h U challenged cells revealed the deposition of uranyl precipitates due to biomineralization. The loss of intracellular structures together with surface and subcellular localization of uranyl precipitates in 24 h U exposed cells were visualized by transmission electron microscopy. Cells treated with 50 µM U exhibited membrane permeabilization which was higher at 200 µM U. Enhanced reactive oxygen species (ROS) accumulation and lipid peroxidation, transient RNA degradation and protein oxidation were observed in U exposed cells. High superoxide dismutase levels coupled with uranium binding and bioprecipitation possibly helped in counteracting U stress in 50 µM U treated cells. Resistance to U toxicity apparently developed under prolonged uranyl (50 µM) incubations. However, cells could not cope up with toxicity at 200 µM U due to impairment of resistance mechanisms.

A novel Sphingobacterium sp. RB, a rhizosphere isolate degrading para-nitrophenol with substrate specificity towards nitrotoluenes and nitroanilines.

Sphingobacterium sp. RB, a novel bacterial strain isolated from a soil sample, was able to utilize para-nitrophenol (PNP) as sole source of carbon and energy at high concentrations (1.0-5.0 mM). The culture completely degraded 3.0 mM PNP within 36 h with proportionate increase in biomass. With 5.0 mM PNP (700 ppm), 70% degradation was observed within 72 h of incubation. Scanning electron microscope images of the isolate in the presence and absence of PNP showed no significant morphological variations. Liquid chromatography-mass spectrometry analysis indicated that the biodegradation of PNP in this bacterium proceeded via the formation of 1,2,4-benzenetriol. Cells previously exposed to PNP (induced) were 30% more effective in degrading PNP. With catechol and phenol, such induction was not observed. Uninduced cells of Sphingobacterium sp. RB were capable of degrading a variety of other nitroaromatic compounds, including 2-nitroaniline, 2,4-dinitroaniline, 2-nitrotoluene, 3-nitrotoluene and 2,4-dinitrophenol, within 72 h, thus proving its candidacy as a potent bioremediation agent. To the best of our knowledge, this is the first report on a Sphingobacterium species degrading PNP via formation of 1,2,4-benzenetriol.

Harnessing the catabolic versatility of Gordonia species for detoxifying pollutants.

The genus Gordonia includes variedly pigmented aerobic, non-motile, non-sporulating Gram positive (sometimes variable) coccoid forms and rods. Different isolates display distinguishing physiological traits and biochemical properties that are significant in remediation applications. Strains inherently prevalent in soils, seawater, sediments and wastewaters can degrade hydrocarbons. Immobilized cells and microbial consortia containing Gordonia species have been used for in situ applications. Hydrocarbon uptake in this Actinomycete is mediated by attachment to large droplets or by pseudosolubilization of substrates. Hydrocarbons so internalized are degraded by relevant enzymes that are innately present in this microorganism. Wild-type and recombinant strains also mediate desulfurization of polyaromatic sulfur heterocyclic compounds. This organism is metabolically capable of bringing about detoxification of phthalate esters. Two species namely, Gordonia polyisoprenivorans and Gordonia westfalica mediate degradation of rubber and the metabolic pathways involved in the process have been well-understood. Some members are able to transform nitriles into commercially valuable products and others degrade the explosive hexahydro-1,3,5-trinitro-1,3,5-triazine. Cholesterol, pyridine derivatives, fuel oxygenates, thiodiglycol, bis-(3-pentafluorophenylpropyl)-sulfide and 6:2 fluorotelomersulfonate are also biotransformed or degraded by Gordonia species. Some members of this genus are significant in the treatment of wastewaters including those that are rich in steroids and lignin. There are also several patents highlighting the commercial relevance of this genus. On account of its diverse catabolic properties, this Actinomycete has become important in bioremediation of polluted environments.

Responses exhibited by various microbial groups relevant to uranium exposure.

There is a strong interest in knowing how various microbial systems respond to the presence of uranium (U), largely in the context of bioremediation. There is no known biological role for uranium so far. Uranium is naturally present in rocks and minerals. The insoluble nature of the U(IV) minerals keeps uranium firmly bound in the earth's crust minimizing its bioavailability. However, anthropogenic nuclear reaction processes over the last few decades have resulted in introduction of uranium into the environment in soluble and toxic forms. Microbes adsorb, accumulate, reduce, oxidize, possibly respire, mineralize and precipitate uranium. This review focuses on the microbial responses to uranium exposure which allows the alteration of the forms and concentrations of uranium within the cell and in the local environment. Detailed information on the three major bioprocesses namely, biosorption, bioprecipitation and bioreduction exhibited by the microbes belonging to various groups and subgroups of bacteria, fungi and algae is provided in this review elucidating their intrinsic and engineered abilities for uranium removal. The survey also highlights the instances of the field trials undertaken for in situ uranium bioremediation. Advances in genomics and proteomics approaches providing the information on the regulatory and physiologically important determinants in the microbes in response to uranium challenge have been catalogued here. Recent developments in metagenomics and metaproteomics indicating the ecologically relevant traits required for the adaptation and survival of environmental microbes residing in uranium contaminated sites are also included. A comprehensive understanding of the microbial responses to uranium can facilitate the development of in situ U bioremediation strategies.

Coculture induced improved production of biosurfactant by Staphylococcus lentus SZ2: Role in protecting Artemia salina against Vibrio harveyi.

Coculturing microorganisms can lead to enhanced production of bioactive compounds as a result of cross-species or cross-genera interactions. In this study, we demonstrate improved production of the biosurfactant (BS-SLSZ2 with antibiofilm properties) by the marine epibiotic bacterium Staphylococcus lentus SZ2 after cross-genera interactions with an aquaculture pathogen Vibrio harveyi. In cocultures, growth of V. harveyi was completely inhibited and resultant biofilms were exclusively composed of S. lentus. The cell free supernatant (CFS) derived from cocultures displayed improved antibiofilm activity with enhanced contents of BS-SLSZ2 compared to monocultured S. lentus. During coculture experiments, after short periods of incubation (6 and 12 h), 2.3 fold increased production of BS-SLSZ2 was observed. Planktonic growth of V. harveyi was also inhibited after coculturing with S. lentus as evidenced from plate culture-based studies and microscopic observations. The CFS derived from monocultures and cocultures did not display bactericidal activity and the observed inhibition of V. harveyi could be of competitive nature. During in vivo challenge experiments, S. lentus protected the model aquaculture system Artemia salina from V. harveyi infections. Seven days post infection, survival of the group of larvae infected with V. harveyi was 5 ±â€¯4.47%. Better survival rates (73.33 ±â€¯5.16%, comparable with the unexposed group) were observed in the group of larvae incubated with S. lentus and V. harveyi. This study highlights increased biosurfactant production by cocultured S. lentus and the application of this bacterium as a protective probiotic strain for inclusion in aquaculture practices.

Morphological response of Yarrowia lipolytica under stress of heavy metals.

The marine dimorphic yeast Yarrowia lipolytica has been proposed as a suitable model for the dimorphism study. In this study, the morphological behaviour of two marine strains of Y. lipolytica (NCIM 3589 and NCIM 3590) was studied under stress of different heavy metals. Scanning electron microscopy was used to investigate the morphological features of yeast cells. This study revealed that the normal ellipsoidal shape of yeast cells was changed into oval, rounded, or elongated in response to different heavy-metal stress. Light microscopy was also used to investigate individual properties of yeast cells. The average cell length and radius of both marine strains was increased with increasing concentrations of heavy-metal ions. In addition, the elongation factor was calculated and was increased in the presence of heavy metals like Pb(II), Co(II), Cr(III), Cr(VI), and Zn(II) under the static conditions.

Heavy metal tolerance in marine strains of Yarrowia lipolytica.

Heavy metal tolerance of two marine strains of Yarrowia lipolytica was tested on solid yeast extract peptone dextrose agar plates. Based on minimum inhibitory concentration esteems, it is inferred that the two strains of Y. lipolytica were tolerant to heavy metals such as Pb(II), Cr(III), Zn(II), Cu(II), As(V), and Ni(II) ions. The impact of various heavy metal concentrations on the growth kinetics of Y. lipolytica was likewise assessed. With increased heavy metal concentration, the specific growth rate was reduced with delayed doubling time. Furthermore, biofilm development of both yeasts on the glass surfaces and in microtitre plates was assessed in presence of different heavy metals. In microtitre plates, a short lag phase of biofilm formation was noticed without the addition of heavy metals in yeast nitrogen base liquid media. A lag phase was extended over increasing metal concentrations of media. Heavy metals like Cr(VI), Cd(II), and As(V) are contrastingly influenced on biofilms' formation of microtitre plates. Other heavy metals did not much influence on biofilms development. Thus, biofilm formation is a strategy of Y. lipolytica under stress of heavy metals has significance in bioremediation process for recovery of heavy metals from contaminated environment.