Biocatalytic lipoprotein bioamphiphile induced treatment of recalcitrant hydrocarbons in petroleum refinery oil sludge through transposon technology.

The present investigation employed transposon technology to enhance the degradation of recalcitrant petroleum hydrocarbons present in petroleum oil sludge by using biosurfactant hyper-producing strain Enterobacter xiangfangensis STP-3. Out of 2500 transposon induced mutants, mutants M257E.xiangfangensis and M916E.xiangfangensis hyper-produce biocatalytic lipoprotein biosurfactant by1.98 and 2.34 fold higher than wild-type strain. Transposon induced mutation also modified the amino acid composition which improved the hydrophobicity and thermal stability of the biosurfactants produced by mutants, compared to the wild-type biosurfactant. GC-MS and LC-MS-MS revealed that biosurfactants have pentameric lipid moiety and esterase as protein moiety. Increased biosurfactant hydrophobicity and yield by the mutants resulted in the enhanced bioavailability of petroleum hydrocarbons, thereby mutants M257E.xiangfangensis and M916E.xiangfangensis demonstrated better petroleum oil sludge degradation by 82% and 88% respectively, than wild-type (72%). Disrupted genes vgr G and pgm M in M257E.xiangfangensis and M916E.xiangfangensis respectively hyper-produce biosurfactant by competitive pathway inhibition and increased precursor availability mechanism. Hyper-production of biosurfactant was also validated by comparing the expression of biosynthetic genes ent E, ent F and est using qPCR. This is the first report on the application of transposon technology to hyper-produce biosurfactant for the effective bioremediation of hydrocarbon contaminated environments.

Biosequestration of lignin in municipal landfill leachate by tailored cationic lipoprotein biosurfactant through Bacillus tropicus valorized tannery solid waste.

Bioremediation of municipal landfill leachate (MLL) is often intricate due to presence of refractory lignin. In the present study, it was attempted to tailor the histidine rich protein moiety of cationic lipoprotein biosurfactant (CLB) to sequester the lignin from MLL. Animal fleshing (AF), the solid waste generated in tanning industry was utilized for the production of histidine rich CLB by de novo substrate dependent synthesis pathway involving Bacillus tropicus. The optimum conditions for the maximum production of CLB were determined using response surface methodology. At the optimized conditions, the maximum yield of CLB was 217.4 mg/g AF (on dry basis). The produced histidine rich CLB was purified using Immobilized metal affinity chromatography at the optimum binding and elution conditions. The histidine residues were more pronounced in the CLB, as determined by HPLC analysis. The CLB was further characterized by SDS-PAGE, Zeta potential, XRD, FT-IR, Raman, NMR, GC-MS and TG analyses. The CLB was immobilized onto functionalized nanoporous activated bio carbon (FNABC) and the optimum immobilization capacity was found to be 211.6 mg/g FNABC. The immobilization of CLB onto FNABC was confirmed using SEM, FT-IR, XRD and TG analyses. The isotherm models, kinetic and thermodynamics studies of CLB immobilization onto FNABC were performed to evaluate its field level application. Subsequently, the CLB-FNABC was then applied for the sequestration of lignin in MLL. The maximum lignin sequestration was achieved by 92.5 mg/g CLB-FNABC at the optimized sequestration time, 180 min; pH, 5; temperature, 45 °C and mass of CLB-FNABC, 1.0 g. The sequestration of lignin by CLB- FNABC was confirmed by SEM, FT-IR and UV-Vis analyses. Further, the mechanistic study revealed the anchoring of CLB onto the surface of lignin through electrostatic interaction.

Preformed aerenchyma determines the differential tolerance response under partial submergence imposed by fresh and saline water flooding in rice.

Plant's response to fresh- and saline-water flooding and the resulting partial submergence, seems different due to the added complexities of element toxicity of salinity. We identified a few rice genotypes which can tolerate combined stresses of partial submergence and salinity during saline water flooding. To gain mechanistic insights, we compared two rice genotypes: Varshadhan (freshwater-flooding tolerant) and Rashpanjor (both fresh- and saline-water flooding tolerant). We found greater ethylene production and increased "respiratory burst oxidase homolog" (RBOH)-mediated reactive oxygen species (ROS) production led to well-developed constitutive aerenchyma formation in Rashpanjor, which makes it preadapted to withstand fresh- and saline-water flooding. On the contrary, an induced aerenchyma formation-dependent tolerance mechanism of Varshadhan worked well for freshwater flooding but failed to provide tolerance to saline-water flooding. Additional salt stress was found to significantly inhibit the induced aerenchyma formation process due to the dampening of ROS signaling by the action of metallothionein in Varshadhan. Besides, inconspicuous changes in ionic regulation processes in these two genotypes under saline-water flooding suggest preadapted constitutive aerenchyma formation plays a more significant role than elemental toxicity per se in tolerating combined stresses encountered during saline water flooding in rice. Overall, our study indicated that well-developed constitutive aerenchyma provide an adaptive advantage during partial submergence due to saline water flooding in rice as the key process of induced aerenchyma formation is hampered in the presence of salinity stress coupled with partial submergence.

Recent Advances of Genetic Resources, Genes and Genetic Approaches for Flooding Tolerance in Rice.

Flooding is one of the most hazardous natural disasters and a major stress constraint to rice production throughout the world, which results in huge economic losses. The frequency and duration of flooding is predicted to increase in near future as a result of global climate change. Breeding of flooding tolerance in rice is a challenging task because of the complexity of the component traits, screening technique, environmental factors and genetic interactions. A great progress has been made during last two decades to find out the flooding tolerance mechanism in rice. An important breakthrough in submergence research was achieved by the identification of major quantitative trait locus (QTL) SUB1 in rice chromosomes that acts as the primary contributor for tolerance. This enabled the use of marker-assisted backcrossing (MABC) to transfer SUB1 QTL into popular varieties which showed yield advantages in flood prone areas. However, SUB1 varieties are not always tolerant to stagnant flooding and flooding during germination stage. So, gene pyramiding approach can be used by combining several important traits to develop new breeding rice lines that confer tolerances to different types of flooding. This review highlights the important germplasm/genetic resources of rice to different types of flooding stress. A brief discussion on the genes and genetic mechanism in rice exhibited to different types of flooding tolerance was discussed for the development of flood tolerant rice variety. Further research on developing multiple stresses tolerant rice can be achieved by combining SUB1 with other tolerance traits/genes for wider adaptation in the rain-fed rice ecosystems.

Rice with SUB1 QTL possesses greater initial leaf gas film thickness leading to delayed perception of submergence stress.

BACKGROUND AND AIMS: Submergence tolerance in rice is primarily attributed to the action of the SUB1 gene, but other associated traits such as leaf gas film (LGF) thickness, leaf hydrophobicity, porosity and leaf density have been known to aid submergence tolerance in rice. However, association of these traits with SUB1 quantitative trait locus (QTL) has not been demonstrated. In this study, we aim to investigate (1) whether the presence of the SUB1 QTL in the genetic background has any influence on the thickness of the LGF and (ii) whether its removal has any impact on stress perception and submergence tolerance in Sub1 and non-Sub1 rice. METHODS: We examined 12 genotypes (including both Sub1 and non-Sub1 types) for different leaf traits such as initial LGF thickness, leaf hydrophobicity, tissue porosity and leaf density in order to work out the relatioship of these traits to the SUB1 QTL in rice. Furthermore, we investigated the changes in the gene expression profile and different metabolic processes in selected genotypes in the presence and absence of their LGF to study its impact on stress perception and adaptation. KEY RESULTS: The initial thickness of the LGF and hydrophobicity seemed to have a highly positive correlation with the presence of the SUB1 QTL in the genetic background of rice; however, other leaf traits such as porosity and density seemed to be independent of it. Artificial removal of the LGF resulted in partial loss of tolerance, showing increased ethylene production and early induction of anoxia-related genes (SUB1A-1, ACS5, Ramy3D and ADH1) which manifested symptoms such as increased stem elongation, faster chlorophyll and starch breakdown, and partial loss of quiescence in SUB1-containing rice genotypes. Stripping of the LGF resulted in early and enhanced induction of SUB1A-1, indicating a quicker perception of stress. CONCLUSIONS: The presence of SUB1 in the genetic background positively influences surface hydrophobicity and the concomitant LGF thickness of rice. Furthermore, LGF helps in terms of providing better ethylene dissipation and reduced in planta accumulation, owing to the slowing down of ethylene-induced leaf senescence under submergence stress.

Biodegradation of petroleum refining industry oil sludge by microbial-assisted biocarrier matrix: process optimization using response surface methodology.

Safe disposal of petroleum oil sludge generated from crude oil storage tank bottom is a major challenge for petroleum refineries across the globe. The presence of long chain hydrocarbons in petroleum oil sludge are known to have effects on the environment through bioaccumulation or biosorption. The present study was focused to develop a modified bioremediation process using hydrocarbonoclastic microbial-assisted biocarrier matrix (MABC) mediated through biosurfactants and biocatalysts for the efficient treatment of petroleum industrial oily sludge. The development of hydrocarbonoclastic microbial-assisted biocarrier matrix was confirmed by scanning electron microscopy analysis. The biocatalysts such as lipase, laccase, esterase and biosurfactant produced by MABC system were found to be 40 U/mg, 18 U/mg, 36 U/mg and 220 mg/g of oil sludge respectively using one variable at a time approach. Further, the response surface methodology was used to determine the optimum treatment conditions (Time, pH, Mass of biocarrier matrix and Amount of oil sludge) for the enhanced removal of total petroleum hydrocarbons (TPH) present in the oil sludge and TPH was degraded by 88.78% at Hydraulic Retention Time of 7 days. The biodegradation of oil sludge was confirmed using Gas Chromatography-Mass Spectrometry analysis.

Tissue Tolerance Coupled With Ionic Discrimination Can Potentially Minimize the Energy Cost of Salinity Tolerance in Rice.

Salinity is one of the major constraints in rice production. To date, development of salt-tolerant rice cultivar is primarily focused on salt-exclusion strategies, which incur greater energy cost. The present study aimed to evaluate a balancing strategy of ionic discrimination vis-à-vis tissue tolerance, which could potentially minimize the energy cost of salt tolerance in rice. Four rice genotypes, viz., FL478, IR29, Kamini, and AC847, were grown hydroponically and subjected to salt stress equivalent to 12 dS m-1 at early vegetative stage. Different physiological observations (leaf chlorophyll content, chlorophyll fluorescence traits, and tissue Na+ and K+ content) and visual scoring suggested a superior Na+-partitioning strategy operating in FL478. A very low tissue Na+/K+ ratio in the leaves of FL478 after 7 days of stress hinted the existence of selective ion transport mechanism in this genotype. On the contrary, Kamini, an equally salt-tolerant genotype, was found to possess a higher leaf Na+/K+ ratio than does FL478 under similar stress condition. Salt-induced expression of different Na+ and K+ transporters indicated significant upregulation of SOS, HKT, NHX, and HAK groups of transporters in both leaves and roots of FL478, followed by Kamini. The expression of plasma membrane and vacuolar H+ pumps (OsAHA1, OsAHA7, and OsV-ATPase) were also upregulated in these two genotypes. On the other hand, IR29 and AC847 showed greater salt susceptibility owing to excess upward transport of Na+ and eventually died within a few days of stress imposition. But in the "leaf clip" assay, it was found that both IR29 and Kamini had high tissue-tolerance and chlorophyll-retention abilities. On the contrary, FL478, although having higher ionic-discrimination ability, showed the least degree of tissue tolerance as evident from the LC50 score (amount of Na+ required to reduce the initial chlorophyll content to half) of 336 mmol g-1 as against 459 and 424 mmol g-1 for IR29 and Kamini, respectively. Overall, the present study indicated that two components (ionic selectivity and tissue tolerance) of salt tolerance mechanism are distinct in rice. Unique genotypes like Kamini could effectively balance both of these strategies to achieve considerable salt tolerance, perhaps with lesser energy cost.

Ionic selectivity and coordinated transport of Na+ and K+ in flag leaves render differential salt tolerance in rice at the reproductive stage.

MAIN CONCLUSION: The present study shows that salt tolerance in the reproductive stage of rice is primarily governed by the selective Na+ and K+ transport from the root to upper plant parts. Ionic discrimination at the flag leaf, governed by differential expression of Na+- and K+-specific transporters/ion pumps, is associated with reduced spikelet sterility and reproductive stage salt tolerance. Reproductive stage salt tolerance is crucial in rice to guarantee yield under saline condition. In the present study, differential ionic selectivity and the coordinated transport (from root to flag leaf) of Na+ and K+ were investigated to assess their impact on reproductive stage salt tolerance. Four rice genotypes having differential salt sensitivity were subjected to reproductive stage salinity stress in pots. The selective Na+ and K+ transport from the root to upper plant parts was observed in tolerant genotypes. We noticed that prolonged salt exposure did not alter flag leaf greenness even up to 6 weeks; however, it had a detrimental effect on panicle development especially in the salt-susceptible genotype Sabita. But more precise chlorophyll fluorescence imaging analysis revealed salinity-induced damages in Sabita. The salt-tolerant genotype Pokkali (AC41585), a potential Na+ excluder, managed to sequester higher Na+ load in the roots with little upward transport as evident from greater expression of HKT1 and HKT2 transporters. In contrast, the moderately salt-tolerant Lunidhan was less selective in Na+ transport, but possessed a higher capacity to Na+ sequestration in leaves. Higher K+ uptake and tissue-specific redistribution mediated by HAK and AKT transporters showed robust control in selective K+ movement from the root to flag leaf and developing panicles. On the contrary, expressions of Na+-specific transporters in developing panicles were either down-regulated or unaffected in tolerant and moderately tolerant genotypes. Yet, in the panicles of the susceptible genotype Sabita, some of the Na+-specific transporter genes (SOS1, HKT1;5, HKT2;4) were upregulated. Apart from the ionic regulation strategy, cellular energy balance mediated by different plasma-membrane and tonoplastic H+-pumps were also associated with the reproductive stage salt tolerance in rice.

Enhanced biodegradation of hydrocarbons in petroleum tank bottom oil sludge and characterization of biocatalysts and biosurfactants.

Petroleum hydrocarbon removal from tank bottom oil sludge is a major issue due to its properties. Conventional physicochemical treatment techniques are less effective. Though the bioremediation is considered for the hydrocarbon removal from tank bottom oil sludge, the efficiency is low and time taking due to the low yield of biocatalysts and biosurfactants. The focal theme of the present investigation is to modify the process by introducing the intermittent inoculation for the enhanced biodegradation of hydrocarbons in the tank bottom oil sludge by maintaining a constant level of biocatalysts such as oxidoreductase, catalase, and lipase as well as biosurfactants. In addition, the heavy metal removal was also addressed. The microbial consortia comprising Shewanalla chilikensis, Bacillus firmus, and Halomonas hamiltonii was used for the biodegradation of oil sludge. One variable at a time approach was used for the optimum of culture conditions. The bacterial consortia degraded the oil sludge by producing biocatalysts such as lipase (80 U/ml), catalase (46 U/ml), oxidoreductase (68 U/ml) along with the production of lipoprotein biosurfactant (152 mg/g of oil sludge) constantly and achieved 96% reduction of total petroleum hydrocarbon. The crude enzymes were characterized by FT-IR and the biosurfactant was characterized by surface tension reduction, emulsification index, FT-IR, TLC, and SDS-PAGE. GC-MS and NMR also revealed that the hydrocarbons present in the oil sludge were effectively degraded by the microbial consortia. The ICP-OES result indicated that the microbial consortium is also effective in removing the heavy metals. Hence, bioremediation using the hydrocarbonoclastic microbial consortium can be considered as an environmentally friendly process for disposal of tank bottom oil sludge from petroleum oil refining industry.

Riboflavin (Vitamin B2) mediated defence induction against bacterial leaf blight: probing through chlorophyll a fluorescence induction O-J-I-P transients.

Bacterial leaf blight (BLB) is a serious threat for rice (Oryza sativa L.) cultivation caused by the bacterial pathogen Xanthomonas oryzae pv. oryzae. The pathogen mainly damages the leaf chlorophyllous tissue, resulting in poor photosynthesis and causing up to 50% reductions in grain yield. In the present work, we have compared the structural and functional ability of the chloroplast of three varieties of rice with different degrees of susceptibility (TN1, highly susceptible; IR-20, moderately resistant; DV-85, resistant to BLB) treated with riboflavin (1 and 2mM) and infected with BLB, with chlorophyll fluorescence as a tool. As indicated by the chlorophyll fluorescence technique, the disease progress curve and yield data, riboflavin acted as an effective vitamin for inducing resistance against BLB. Plants treated with riboflavin showed improved PSII activity, more chlorophyll content and higher yield than the diseased plants.

GC-MS Analysis of Bio-active Molecules Derived from Paracoccus pantotrophus FMR19 and the Antimicrobial Activity Against Bacterial Pathogens and MDROs.

The present investigation is focused on the study of chemical composition of a bioactive compound derived from a rumen isolate Paracoccus pantotrophus FMR19 using GC-MS and to find out the antibacterial activity of the extracted crude bioactive compounds against multidrug resistant organisms (MDROs) and other clinical pathogens. GC-MS analysis revealed that P. pantotrophus FMR19 produced eight major compounds that have been reported to exhibit antimicrobial property. The main components identified from hexane fraction are long chain alkanes, fatty alcohols, fatty acid methyl ester and aromatic hydrocarbons. These molecules are not only active against clinical pathogens such as Salmonella sp. and Proteus sp. and also effective against MDROs such as Metallo ß lactamase and Pan drug resistant bacterial strains and Methicillin resistant Staphylococcus aureus.

Do the pre-service education programmes for midwives in India prepare confident 'registered midwives'? A survey from India.

Objective The graduates of the diploma and degree programmes of nursing and midwifery in India are considered skilled birth attendants (SBAs). This paper aimed to assess the confidence of final-year students from pre-service education programmes (diploma and bachelor's) in selected midwifery skills from the list of midwifery competencies of the International Confederation of Midwives (ICM). Design A cross-sectional survey was conducted in Gujarat, India, involving 633 final-year students from 25 educational institutions (private or government), randomly selected, stratified by the type of programme (diploma and bachelor's). Students assessed their confidence on a four-point scale, in four midwifery competency domains - antepartum, intrapartum, postpartum, and newborn care. Explorative factor analysis was used to reduce skill statements into separate subscales for each domain. Results Overall, 25-40% of students scored above the 75th percentile and 38-50% below the 50th percentile of confidence in all subscales for antepartum, intrapartum, postpartum, and newborn care. The majority had not attended the required number of births prescribed by the Indian Nursing Council. Conclusions The pre-service education offered in the diploma and bachelor's programmes in Gujarat does not prepare confident SBAs, as measured on selected midwifery competencies of the ICM. One of the underlying reasons was less clinical experience during their education. The duration, content, and pedagogy of midwifery education within the integrated programmes need to be reviewed.

Using 'appreciative inquiry' in India to improve infection control practices in maternity care: a qualitative study.

BACKGROUND: Infections acquired during childbirth are a common cause of maternal and perinatal mortality and morbidity. Changing provider behaviour and organisational settings within the health system is key to reducing the spread of infection. OBJECTIVE: To explore the opinions of health personnel on health system factors related to infection control and their perceptions of change in a sample of hospital maternity units. DESIGN: An organisational change process called 'appreciative inquiry' (AI) was introduced in three maternity units of hospitals in Gujarat, India. AI is a change process that builds on recognition of positive actions, behaviours, and attitudes. In-depth interviews were conducted with health personnel to elicit information on the environment within which they work, including physical and organisational factors, motivation, awareness, practices, perceptions of their role, and other health system factors related to infection control activities. Data were obtained from three hospitals which implemented AI and another three not involved in the intervention. RESULTS: Challenges which emerged included management processes (e.g. decision-making and problem-solving modalities), human resource shortages, and physical infrastructure (e.g. space, water, and electricity supplies). AI was perceived as having a positive influence on infection control practices. Respondents also said that management processes improved although some hospitals had already undergone an accreditation process which could have influenced the changes described. Participants reported that team relationships had been strengthened due to AI. CONCLUSION: Technical knowledge is often emphasised in health care settings and less attention is paid to factors such as team relationships, leadership, and problem solving. AI can contribute to improving infection control by catalysing and creating forums for team building, shared decision making and problem solving in an enabling environment.

Coherence between health policy and human resource strategy: lessons from maternal health in Vietnam, India and China.

The failure to meet health goals such as the Millennium Development Goals (MDG) is partly due to the lack of appropriate resources for the effective implementation of health policies. The lack of coherence between the health policies and human resource (HR) strategy is one of the major causes. This article explores the relationship and the degree of coherence between health policy--in this case maternal health policy--processes and HR strategy in Vietnam, China and India in the period 2005-09. Four maternal health policy case studies were explored [skilled birth attendance (SBA), adolescent and sexual reproductive health, domestic violence and medical termination of pregnancy] across three countries through interviews with key respondents, document analysis and stakeholder meetings. Analysis for coherence between health policy and HR strategy was informed by a typology covering 'separation', 'fit' and 'dialogue'. Regarding coherence we found examples of complete separation between health policy and HR strategy, a good fit with the SBA policy though modified through 'dialogue' in Vietnam, and in one case a good fit between policy and strategy was developed through successive evaluations. Three key influences on coherence between health policy and HR strategy emerge from our findings: (1) health as the lead sector, (2) the nature of the policy instrument and (3) the presence of 'HR champions'. Finally, we present a simple algorithm to ensure that appropriate HR related actors are involved; HR is considered at the policy development stage with the option of modifying the policy if it cannot be adequately supported by the available health workforce; and ensuring that HR strategies are monitored to ensure continued coherence with the health policy. This approach will ensure that the health workforce contributes more effectively to meeting the MDGs and future health goals.

The effect of surveillance and appreciative inquiry on puerperal infections: a longitudinal cohort study in India.

OBJECTIVE: To evaluate the effects of an intervention comprising surveillance and an organisational change called Appreciative Inquiry on puerperal infections in hospitals in Gujarat state, India. METHODS: This longitudinal cohort study with a control group was conducted over 16 months between 2010 and 2012. Women who delivered in six hospitals were followed-up. After a five month pre-intervention period, the intervention was introduced in three hospitals. Monthly incidence of puerperal infection was recorded throughout the study in all six hospitals. A chi-square test and logistic regression were used to examine for associations, trends and interactions between the intervention and control groups. FINDINGS: Of the 8,124 women followed up, puerperal infections were reported in 319 women (3.9%) over the course of the study. Puerperal sepsis/genital tract infections and urinary tract infections were the two most common puerperal infections. At the end of the study, infection incidence in the control group halved from 7.4% to 3.5%. Levels in the intervention group reduced proportionately even more, from 4.3% to 1.7%. A chi-square test for trend confirmed the reduction of infection in the intervention and control groups (p<0.0001) but the trends were not statistically different from one another. There was an overall reduction of infection by month (OR = 0.94 95% CI 0.91-0.97). Risk factors like delivery type, complications or delivery attendant showed no association with infection. CONCLUSION: Interruption of resource flows in the health system occurred during the intervention phase, which may have affected the findings. The incidence of infection fell in both control and intervention groups during the course of the study. It is not clear if appreciative inquiry contributed to the reductions observed. A number of practical and methodological limitations were faced. TRIAL REGISTRATION: Controlled-Trials.com ISRCTN03513186.

Lipase from marine strain using cooked sunflower oil waste: production optimization and application for hydrolysis and thermodynamic studies.

The marine strain Pseudomonas otitidis was isolated to hydrolyze the cooked sunflower oil (CSO) followed by the production of lipase. The optimum culture conditions for the maximum lipase production were determined using Plackett-Burman design and response surface methodology. The maximum lipase production, 1,980 U/ml was achieved at the optimum culture conditions. After purification, an 8.4-fold purity of lipase with specific activity of 5,647 U/mg protein and molecular mass of 39 kDa was obtained. The purified lipase was stable at pH 5.0-9.0 and temperature 30-80 °C. Ca(2+) and Triton X-100 showed stimulatory effect on the lipase activity. The purified lipase was highly stable in the non-polar solvents. The functional groups of the lipase were determined by Fourier transform-infrared (FT-IR) spectroscopy. The purified lipase showed higher hydrolytic activity towards CSO over the other cooked oil wastes. About 92.3 % of the CSO hydrolysis was observed by the lipase at the optimum time 3 h, pH 7.5 and temperature 35 °C. The hydrolysis of CSO obeyed pseudo first order rate kinetic model. The thermodynamic properties of the lipase hydrolysis were studied using the classical Van't Hoff equation. The hydrolysis of CSO was confirmed by FT-IR studies.

Microbial induced lipoprotein biosurfactant from slaughterhouse lipid waste and its application to the removal of metal ions from aqueous solution.

This study aims at demonstrating the production of lipoprotein biosurfactant from Pseudomonas gessardii using goat tallow, a slaughterhouse lipid waste, as the substrate and its application to the removal of metal ions from aqueous solution. The maximum bio-transformation of goat tallow into biosurfactant occurred at 48 h. The mass of the lipoprotein biosurfactant produced was 2.03 g/g of goat tallow. The biosurfactant was clearly characterized by surface tension, critical micelle concentration, emulsification index and molecular weight. The amino acid and fatty acid moieties of the biosurfactant were determined using High performance liquid chromatography (HPLC) and Gas chromatography (GC). The thermal behavior studies were evaluated using Thermo gravimetric (TG) and Differential scanning calorimetry (DSC) analysis. The lipoprotein biosurfactant was loaded onto the mesoporous activated carbon (MAC) for the sequestering of metal ions from the aqueous solution. The biosurfactant exhibited a removal efficiency for metal ions from aqueous solution in the order Cr(3+)>Ca(2+)>Cu(2+)>Fe(2+). The morphological observations and functional groups of the lipoprotein biosurfactant and that of the lipoprotein biosurfactant bound metal ions were determined using scanning electron micrograph (SEM) images and Fourier transform infrared (FT-IR) spectroscopy, respectively. This is the first report on the production of lipoprotein biosurfactant by P. gessardii using goat tallow as the substrate to sequester the metal ions from the aqueous solution.

Production of lipase from Pseudomonas gessardii using blood tissue lipid and thereof for the hydrolysis of blood cholesterol and triglycerides and lysis of red blood cells.

The study demonstrates the production of lipase (LIP) from Pseudomonas gessardii using blood tissue lipid as the substrate for the hydrolysis of blood cholesterol and triglycerides. The lipase was purified with the specific activity of 828 U/mg protein and the molecular weight of 56 kDa. The maximum lipase activity was observed at the pH 7.0 and the temperature 37 °C. The amino acid composition of purified lipase was determined by HPLC. The mesoporous activated carbon (MAC) was used for the immobilization of lipase for the repeated use of the enzyme catalyst. The K (m) value of immobilized lipase (MAC-LIP) and the free lipase (LIP) was 0.182 and 1.96 mM, respectively. The V (max) value of MAC-LIP and LIP was 1.33 and 1.26 mM/min, respectively. The MAC and MAC-LIP were characterized by scanning electron microscopy (SEM). The hydrolysis study showed 78 and 100% hydrolysis of triglycerides and cholesterol, respectively, for LIP and 84 and 100% hydrolysis of triglycerides and cholesterol, respectively, for MAC-LIP at the reaction time of 1 h. The effect of lipase on cell wall lysis was carried out on the RBCs of blood plasma. Interestingly, 99.9% lysis of RBCs was observed within 2 h. SEM images and phase contrast microscopy confirmed the lysis of RBCs. This work provides a potential biocatalyst for the hydrolysis of blood cholesterol and triglycerides.