Genome-centric polyhydroxyalkanoate reconciliation reveals nutrient enriched growth dependent biosynthesis in Bacillus cereus IBA1.

Microbiological polyhydroxyalkanoates (PHAs) are rooted as the most promising bio-replacements of synthetic polymers. Inherent properties of these PHAs further expand their applicability in numerous industrial, environmental, and clinical sectors. To propel these, a new environmental, endotoxin free gram-positive bacterium i.e., Bacillus cereus IBA1 was identified to harbor advantageous PHA producer characteristics through high-throughput omics mining approaches. Unlike traditional fermentations, nutrient enriched strategy was used to enhance PHA granular concentrations by ∼2.3 folds to 2.78 ± 0.19 g/L. Additionally, this study is the first to confirm an underlying growth dependent PHA biogenesis through exploring PHA granule associated operons which harbour constitutively expressing PHA synthase (phaC) coupled with differentially expressing PHA synthase subunit (phaR) and regulatory protein (phaP, phaQ) amid different growth phases. Moreover, the feasibility of this promising microbial phenomenon could propel next-generation biopolymers, and increase industrial applicability of PHAs, thereby significantly contributing to the sustainable development.

Acclimatization of microbial community of submerged membrane bioreactor treating hospital wastewater.

This study was performed to understand the dynamics of the microbial community of submerged membrane bioreactor during the acclimatization process to treat the hospital wastewater. In this regard, three acclimatization phases were examined using a mixture of synthetic wastewater (SWW) and real hospital wastewater (HWW) in the following proportions; In Phase 1: 75:25 v/v (SWW: HWW); Phase 2: 50:50 v/v (SWW: HWW); and Phase 3: 25:75 v/v (SWW: HWW) of wastewater. The microbial community was analyzed using Illumina high throughput sequencing to identify the bacterial and micro-eukaryotes community in SMBR. The acclimatization study clearly demonstrated that shift in microbial community composition with time. The dominance of pathogenic and degrading bacterial communities such as Mycobacterium, Pseudomonas, and Zoogloea was observed at the phase 3 of acclimatization. This study witnessed the major shift in the micro-eukaryotes community, and the proliferation of fungi Basidiomycota was observed in phase 3 of acclimatization.

Effect of cathode material and charge loading on the nitrification performance and bacterial community in leachate treating Electro-MBRs.

Electro-MBR technology, which combines an electrocoagulation process inside the mixed liquor of a membrane bioreactor, was studied for the treatment of a high-strength ammonia leachate (124 ± 4 mg NH4-N L-1). A lab-scale aerobic Electro-MBR was operated with a solid retention time of 45 days, hydraulic retention times of 24h and 12h, and charge loading ranging from 100 to 400 mAh L-1. At 400 mAh L-1, with a combination of a Ti/Pt cathode and a sacrificial iron anode, removal percentages for ammonia nitrogen, total organic carbon, and total phosphorus were 99.8%, 38%, and 99.0%, respectively. At 400 mAh L-1, the estimated ferric ion dosage was 325 mg Fe3+ L-1. Experiments conducted with different cathode materials showed that previously reported inhibition phenomena may result from a cathodic nitrate reduction into ammonia nitrogen. Conventional cathode materials, such as graphite, have electrochemical nitrate reduction rates of -0.03 mg NO3-N mAh-1. By comparison, when using Ti/Pt, the rate was -0.0045 mg NO3-N mAh-1(85% lower than graphite due to its low hydrogen overpotential). Charge loading tested in this study had no significant impact on both nitrification performance and microbial population diversity. However, the relative abundance of the mixed liquor's Nitrosomonas increased from 4.8% to 8.2% when the charge loading increased from 0 to 400 mAh L-1. Results from this study are promising for future applications of the Ti/Pt - Iron Electro-MBR in various high-strength ammonia wastewater treatment applications.

Identifying the link between MBRs' key operating parameters and bacterial community: A step towards optimized leachate treatment.

A MBR treating compost leachate was studied in order to link the operating parameters (solid and hydraulic retention time) to contaminant's specific bacterial catabolic activity. In this context, a lab-scale aerobic membrane bioreactor was operated for 200 days, at solid retention times (SRT) of 30 and 45 days and four different contaminant load rates. Results showed that increasing the food to microorganism ratio (F/M) by increasing the contaminant load rates lessened the selectivity pressure, which allowed the proliferation of subdominant operational taxonomic units (OTU) (relative abundance >3%) that were otherwise inhibited by highly adapted dominant OTUs (relative abundance >10%). Subsequently, increasing the SRT resulted in a lower species richness and the selection of two dominant types of bacteria: 1) genera with low growth rates that feed on non-limiting substrates or substrates with few competitors, and 2) genera with metabolisms that are highly specific to the available substrates and that can outcompete the other genera by using the substrate more efficiently. The bacterial population evolution observed during this study suggests that the mixed liquor population diversity and structure can be modulated with the operating conditions for the bioenhancement of contaminant specific catabolic activity. Identified dominant and subdominant genera were linked to the MBR's NH4+ and COD removal performances. Interestingly, nitrification performances were unaffected by the organic load rate and the Nitrosomonas relative abundance.

MBR treatment of leachates originating from waste management facilities: A reference study of the design parameters for efficient treatment.

The main objective of the study was to define the interaction between the solid retention time (SRT) and the contaminant loading rate on a membrane bioreactor's efficacy in removing contaminants frequently detected (chemical oxygen demand (COD), NH4+, total phosphorus and metals) above the discharge criteria in waste-originating leachates. The rates and coefficient calculated from this study's experimental data can be used for the design of membrane bioreactor treating wastewaters, even beyond the scope of this experiment. Over a period of 152 days, SRTs of 28 and 47 days and HRTs of 13, 25, 36 and 52 h were studied using a real leachate with a constant composition. Results showed that membrane bioreactors can efficiently treat >1850 mg COD L-1 d-1 of highly to moderately biodegradable COD, with the SRT having no significant impact on the removal of recalcitrant COD. Overall ammonium removal rates of >740 mg NH4-N L-1 d-1 can be achieved as long as a residual alkalinity of 200 mg CaCO3 L-1 and an adequate dissolved oxygen concentration (6-7 mg L-1) are both maintained. Overall phosphorus removal rates are independent of the phosphorus loading rate. However, the highest overall phosphorus removal rate (39 ± 2 mg P per g of total suspended solids) was obtained at the lowest SRT (28 days) due to an increased extracellular polymeric substance production. Finally, membrane bioreactor's metal removal capacity is mostly dependent on the metals' affinity to both the leachate's recalcitrant COD as well as sludge concentrations.

Biopesticide and formulation processes based on starch industrial wastewater fortified with soybean medium.

The aim of this study was to produce Bacillus thuringiensis-based biopesticide using starch-producing industry wastewater (SIW) fortified with soybean medium and optimize the formulated product using different adjuvants. This study was necessary as low endotoxin concentration is obtained in formulated biopesticide when SIW alone is used as fermentation medium. The fermentation runs were conducted using SIW alone and SIW fortified with 25% soybean (w/v) medium in 2000 L and 150 L bioreactor, respectively. SIW supplemented with soybean medium showed an increase in cell count (from 1.95 × 108 to 1.65 × 109 CFU mL-1), spore synthesis (from 1.5 × 108 to 1.35 × 109 CFU mL-1) and endotoxin concentration (from 436 to 1170 µg mL-1) when compared to SIW medium alone. The fermented broth was concentrated using continuous centrifugation and adjuvants were added for biopesticide formulation in order to enhance its resistance against UV rays and rainfastness. Entomotoxicity of the formulation produced using fermented broth of SIW fortified with soybean (38,000 IU µL-1) was higher than that obtained by SIW medium alone (21,000 IU µL-1), commercial biopesticide Foray 76B (20,000 IU µL-1) and Btk sander's (12,500 IU µL-1).

The bacterial community structure of submerged membrane bioreactor treating synthetic hospital wastewater.

The pharmaceuticals are biologically active compounds used to prevent and treat diseases. These pharmaceutical compounds were not fully metabolized by the human body and thus excreted out in the wastewater stream. Thus, the study on the treatment of synthetic hospital wastewater containing pharmaceuticals (ibuprofen, carbamazepine, estradiol and venlafaxine) was conducted to understand the variation of the bacterial community in a submerged membrane bioreactor (SMBR) at varying hydraulic retention time (HRT) of 6, 12 and 18 h. The variation in bacterial community dynamics of SMBR was studied using high throughput sequencing. The removal of pharmaceuticals was uniform at varying HRT. The removal of both ibuprofen and estradiol was accounted for 90%, whereas a lower removal of venlafaxine (<10%) and carbamazepine (>5%) in SMBR was observed. The addition of pharmaceuticals alters the bacterial community structure and result in increased abundance of bacteria (e.g., Flavobacterium, Pedobacter, and Methylibium) reported to degrade toxic pollutant.

Recent developments of downstream processing for microbial lipids and conversion to biodiesel.

With increasing global population and depleting resources, there is an apparent demand for radical unprecedented innovation to satisfy the basal needs of lives. Hence, non-conventional renewable energy resources like biodiesel have been worked out in past few decades. Biofuel (e.g. Biodiesel) serves to be the most sustainable answer to solve "food vs. fuel crisis". In biorefinery process, lipid extraction from oleaginous microbial lipids is an integral part as it facilitates the release of fatty acids. Direct lipid extraction from wet cell-biomass is favorable in comparison to dry-cell biomass because it eliminates the application of expensive dehydration. However, this process is not commercialized yet, instead, it requires intensive research and development in order to establish robust approaches for lipid extraction that can be practically applied on an industrial scale. This review aims for the critical presentation on cell disruption, lipid recovery and purification to support extraction from wet cell-biomass for an efficient transesterification.

Pilot-scale biopesticide production by Bacillus thuringiensis subsp. kurstaki using starch industry wastewater as raw material.

Pilot-scale Bacillus thuringiensis based biopesticide production (2000 L bioreactor) was conducted using starch industry wastewater (SIW) as a raw material using optimized operational parameters obtained in 15 L and 150 L fermenters. In pilot scale fermentation process the oxygen transfer rate is a major limiting factor for high product yield. Thus, the volumetric mass transfer coefficient (KLa) remains a tool to determine the oxygen transfer capacity [oxygen utilization rate (OUR) and oxygen transfer rate (OTR)] to obtain better bacterial growth rate and entomotoxicity in new bioreactor process optimization and scale-up. This study results demonstrated that the oxygen transfer rate in 2000 L bioreactor was better than 15 L and 150 L fermenters. The better oxygen transfer in 2000 L bioreactor augmented the bacterial growth [total cell (TC) and viable spore count (SC)] and delta-endotoxin yield. Prepared a stable biopesticide formulation for field use and its entomotoxicity was also evaluated. This study result corroborates the feasibility of industrial scale operation of biopesticide production using starch industry wastewater as raw material.

Review on fate and mechanism of removal of pharmaceutical pollutants from wastewater using biological approach.

Due to research advancement and discoveries in the field of medical science, maintains and provides better human health and safer life, which lead to high demand for production of pharmaceutical compounds with a concomitant increase in population. These pharmaceutical (biologically active) compounds were not fully metabolized by the body and excreted out in wastewater. This micro-pollutant remains unchanged during wastewater treatment plant operation and enters into the receiving environment via the discharge of treated water. Persistence of pharmaceutical compounds in both surface and ground waters becomes a major concern due to their potential eco-toxicity. Pharmaceuticals (emerging micro-pollutants) deteriorate the water quality and impart a toxic effect on living organisms. Therefore, from last two decades, plenty of studies were conducted on the occurrence, impact, and removal of pharmaceutical residues from the environment. This review provides an overview on the fate and removal of pharmaceutical compounds via biological treatment process.

Detergent assisted ultrasonication aided in situ transesterification for biodiesel production from oleaginous yeast wet biomass.

In situ transesterification of oleaginous yeast wet biomass for fatty acid methyl esters (FAMEs) production using acid catalyst, methanol with or without N-Lauroyl sarcosine (N-LS) treatment was performed. The maximum FAMEs yield obtained with or without N-LS treatment in 24h reaction time was 96.1±1.9 and 71±1.4% w/w, respectively. The N-LS treatment of biomass followed by with or without ultrasonication revealed maximum FAMEs yield of 94.3±1.9% and 82.9±1.8% w/w using methanol to lipid molar ratio 360:1 and catalyst concentration 360mM (64µL H2SO4/g lipid) within 5 and 25min reaction time, respectively. The FAMEs composition obtained in in situ transesterification was similar to that obtained with conventional two step lipid extraction and transesterification process. Biodiesel fuel properties (density, kinematic viscosity, cetane number and total glycerol) were in accordance with international standard (ASTM D6751), which suggests the suitability of biodiesel as a fuel.

Detergent assisted lipid extraction from wet yeast biomass for biodiesel: A response surface methodology approach.

The lipid extraction from the microbial biomass is a tedious and high cost dependent process. In the present study, detergent assisted lipids extraction from the culture of the yeast Yarrowia lipolytica SKY-7 was carried out. Response surface methodology (RSM) was used to investigate the effect of three principle parameters (N-LS concentration, time and temperature) on microbial lipid extraction efficiency % (w/w). The results obtained by statistical analysis showed that the quadratic model fits in all cases. Maximum lipid recovery of 95.3±0.3% w/w was obtained at the optimum level of process variables [N-LS concentration 24.42mg (equal to 48mgN-LS/g dry biomass), treatment time 8.8min and reaction temperature 30.2°C]. Whereas the conventional chloroform and methanol extraction to achieve total lipid recovery required 12h at 60°C. The study confirmed that oleaginous yeast biomass treatment with N-lauroyl sarcosine would be a promising approach for industrial scale microbial lipid recovery.

Energy balance of biofuel production from biological conversion of crude glycerol.

Crude glycerol, a by-product of biodiesel production, has gained significant attention as a carbon source for biofuel production. This study evaluated the energy balance of biodiesel, hydrogen, biogas, and ethanol production from 3.48 million L of crude glycerol (80% w/v). The conversion efficiency (energy output divided by energy invested) was 1.16, 0.22, 0.27, and 0.40 for the production of biodiesel, hydrogen, biogas, and ethanol respectively. It was found that the use of crude glycerol for biodiesel production was an energy gain process, with a positive energy balance and conversion efficiency of greater than 1. The energy balance revealed a net energy gain of 5226 GJ per 1 million kg biodiesel produced. Production of hydrogen, biogas and ethanol from crude glycerol were energy loss processes. Therefore, the conversion of crude glycerol to lipids and subsequently to biodiesel is suggested to be a better option compared to hydrogen, biogas, or ethanol production with respect to energy balance.

Corrosion and stability study of Bacillus thuringiensis var. kurstaki starch industry wastewater-derived biopesticide formulation.

Biopesticides are usually sprayed on forests by using planes made up of aluminum alloy. Bioval derived from starch industry wastewater (SIW) in suspension form was developed as stable anticorrosive biopesticide formulation. In this context, various anticorrosion agents such as activated charcoal, glycerin, ethylene glycol, phytic acid, castor oil and potassium silicate were tested as anticorrosive agents. There was no corrosion found in Bioval formulation where potassium silicate (0.5% w/v) was added and compared with Foray 76 B, as an industrial standard, when stored over 6 months. In relation to other parameters, the anticorrosion formulation of Bioval+buffer+KSi reported excellent zeta potential (-33.19 ± 4 mV) and the viscosity (319.13 ± 32 mPa.s) proving it's stability over 6 months, compared to the standard biopesticide Foray 76 B (-36.62 ± 4 mV potential zeta, pH 4.14 ± 0.1 and 206 ± 21 mPa.s viscosity). Metal analysis of the different biopesticides showed that Bioval+buffer+KSi has no corrosion (5.11 ± 0.5 mg kg(-1) of Al and 13.53 ± 1.5 mg kg(-1) of Fe) on the aluminum alloy due to the contribution of sodium acetate buffer at pH 5. The bioassays reported excellent results for Bioval+Buffer+KSi (2.95 ± 0.3 × 10(9) CFU mL(-1) spores and 26.6 ± 2.7 × 10(9) IU L(-1) Tx) compared with initial Bioval (2.46 ± 0.3 × 10(9) CFU mL(-1) spores and 23.09 ± 3 × 10(9) IU L(-1) Tx) and Foray 76 B (2.3 ± 0.2 × 10(9) CFU mL(-1) spores and 19.950 ± 2.1 UI L(-1) Tx) which was due to the break-up of the external chitinous membrane due to abrasive action of potassium silicate after ingestion by insects. The contribution of sodium acetate buffer and potassium silicate (0.5% and at pH = 5) as anticorrosion agent in the Bioval allowed production of an efficient biopesticide with a reduced viscosity and favorable pH as compared to Foray 76 B which enhanced the entomotoxic potential against spruce budworm (SB) larvae (Lepidoptera: Choristoneura fumiferana).

Ultrasonication assisted lipid extraction from oleaginous microorganisms.

Various solvents, including water, hexane, methanol, and chloroform/methanol (1:1 v/v), were tested to identify the efficiency of lipid extraction from Trichosporon oleaginosus and an oleaginous fungal strain SKF-5 under ultrasonication (520 kHz 40 W and 50 Hz 2800 W) and compared with the conventional chloroform methanol (2:1 v/v) extraction method. The highest lipid recovery 10.2% and 9.3% with water, 43.2% and 33.2% with hexane, 75.7% and 65.1% with methanol, 100% and 100% w/w biomass with chloroform/methanol were obtained from T. oleaginosus and SKF-5 strain, respectively, at ultrasonication frequency 50 Hz and power input 2800 W. Ultrasonication chloroform/methanol extraction recovered total lipid in a short time (15 min) and low temperature (25°C). Whereas the conventional chloroform methanol extraction to achieve total lipid recovery required 12h at 60°C. Ultrasonication chloroform/methanol extraction would be a promising method of lipid extraction from the microorganisms.

Removal of estrogenic activity of natural and synthetic hormones from a municipal wastewater: efficiency of horseradish peroxidase and laccase from Trametes versicolor.

Some researches studied the removal of steroid estrogens by enzymatic treatment, however none verified the residual estrogenicity after the enzymatic treatment at environmental conditions. In this study, the residual estrogenic activities of the key natural and synthetic steroid estrogens were investigated following enzymatic treatment with horseradish peroxidase (HRP) and laccase from Trametes versicolor. Synthetic water and municipal wastewater containing environmental concentrations of estrone, 17beta-estradiol, estriol, and 17alpha-ethinylestradiol were treated. Liquid chromatography-mass spectrometry analysis demonstrated that the studied steroid estrogens were completely oxidized in the wastewater reaction mixture after a 1-h treatment with either HRP (8-10 U ml(-1)) or laccase (20 U ml(-1)). Using the recombinant yeast assay, it was also confirmed that both enzymatic treatments were very efficient in removing the estrogenic activity of the studied steroid estrogens. The laccase-catalyzed process seemed to present great advantages over the HRP-catalyzed system for up-scale applications for the treatment of municipal wastewater.

Laccase-catalyzed conversion of natural and synthetic hormones from a municipal wastewater.

The Trametes versicolor-derived laccase-catalyzed oxidation of natural estrogens (estrone--E1; 17beta-estradiol--E2; and estriol--E3) and a synthetic estrogen (17alpha-ethinylestradiol--EE2) was studied in synthetic water and municipal wastewater to optimize the process for steroid estrogen removal in wastewater. The optimal pH for each studied steroid estrogen oxidation was approximately 6 in synthetic water. This research also focused on the wastewater matrix effect on developed enzymatic treatment. At pH 7.0 and 25+/-1 degrees C, the experiments showed that the laccase-catalyzed system for the removal of steroid estrogens was not significantly affected by the municipal wastewater matrix. Laccase activity of 20 U/ml was sufficient to achieve complete removal of studied steroid estrogens in both synthetic water and municipal wastewater. Moreover, 1-hydroxy-benzotriazole, when used as a mediator, improved laccase-catalyzed system efficiency, thus decreasing the overall cost of the enzymatic system.

Oxidation of natural and synthetic hormones by the horseradish peroxidase enzyme in wastewater.

Steroid estrogens, including both natural estrogens (e.g., estrone - E1; 17beta-estradiol - E2; and estriol - E3) and synthetic estrogens (e.g., 17alpha-ethinylestradiol - EE2), are known as endocrine-disrupting compounds. The objective of this research was to evaluate the feasibility of the enzymatic oxidation of estrogens and to optimize this process in municipal wastewater contaminated with steroid estrogens using horseradish peroxidase (HRP) and hydrogen peroxide. An initial HRP activity of 0.02 U ml(-1) was sufficient to completely remove EE2 from the synthetic solution, although greater HRP doses (up to 0.06 U ml(-1)) were required to remove E1, E2 and E3. The optimal molar peroxide-to-substrate ratio was determined to be approximately 0.45. Based on the Michaelis-Menten kinetics, the HRP had an increasing reactivity with E1, E3, E2, and EE2, in increasing order. In real activated sludge process effluent, an HRP dose of 8-10 U ml(-1) was required to completely remove all of the studied estrogens, while only 0.032 U ml(-1) of HRP was necessary to treat synthetic water containing the same estrogen concentrations.

Natural and synthetic hormone removal using the horseradish peroxidase enzyme: temperature and pH effects.

The primary objective of our research was to establish the technical feasibility of using the horseradish peroxidase (HRP) enzyme for natural and synthetic estrogens-estrone (E1), 17beta-estradiol (E2), estriol (E3), and 17alpha-ethinylestradiol (EE2)-removal. The effects of temperature and pH on enzymatic treatment kinetics were investigated. Residual estrogen concentrations were quantified by liquid chromatography, coupled with mass spectrometry analysis. In a synthetic solution at pH 7 and 25+/-1 degrees C, the HRP enzyme-catalyzed process was capable of achieving 92-100% removal of E1, E2, E3, and EE2 within 1h of treatment with an HRP activity of 0.017 U/ml. The influence of the pH (5-9) and temperature (5-35 degrees C) on estrogen removal was observed to be significant, with the optimum pH near neutral conditions. The results also showed that wastewater constituents significantly impact the HRP-catalyzed estrogen removal. The experimental research proved that the HRP-catalyzed system is technically feasible for the removal of the main estrogens present in the environment at low concentrations.