Enhanced biomass production through optimization of carbon source and utilization of wastewater as a nutrient source.

The study aimed to utilize the domestic wastewater as nutrient feedstock for mixotrophic cultivation of microalgae by evaluating appropriate carbon source. The microalgae Chlorella vulgaris was cultivated in municipal wastewater under various carbon sources (glucose, glycerol, and acetate), followed by optimization of appropriate carbon source concentration to augment the biomass, lipid, and carbohydrate contents. Under optimized conditions, namely of 5 g/L glucose, C. vulgaris showed higher increments of biomass with 1.39 g/L dry cell weight achieving biomass productivity of 0.13 g/L/d. The biomass accumulated 19.29 ± 1.83% total lipid, 41.4 ± 1.46% carbohydrate, and 33.06 ± 1.87% proteins. Moreover, the cultivation of Chlorella sp. in glucose-supplemented wastewater removed 96.9% chemical oxygen demand, 65.3% total nitrogen, and 71.2% total phosphate. The fatty acid methyl ester obtained showed higher amount (61.94%) of saturated fatty acid methyl esters associated with the improved fuel properties. These results suggest that mixotrophic cultivation using glucose offers great potential in the production of renewable biomass, wastewater treatment, and consequent production of high-value microalgal oil.

Integration of microalgal cultivation system for wastewater remediation and sustainable biomass production.

Untreated wastewaters have been a great concern and can cause major pollution problems for environment. Conventional approaches for treating wastewater involve tremendous capital cost, have major short comings and are not sustainable. Microalgae culture offers an interesting step for wastewater treatment. Microalgae serve the dual purpose of phycoremediation along with the production of potentially valuable biomass, which can be used for several purposes. The ability of microalgae to accumulate nitrogen, phosphorus, heavy metals and other toxic compounds can be integrated with wastewater treatment system to offer an elegant solution towards tertiary and quaternary treatment. The current review explores possible role of microalgal based wastewater treatment and explores the current progress, key challenges, limitations and future prospects with special emphasis on strategies involved in harvesting, boosting biomass and lipid yield.

A mini review: photobioreactors for large scale algal cultivation.

Microalgae cultivation has gained much interest in terms of the production of foods, biofuels, and bioactive compounds and offers a great potential option for cleaning the environment through CO2 sequestration and wastewater treatment. Although open pond cultivation is most affordable option, there tends to be insufficient control on growth conditions and the risk of contamination. In contrast, while providing minimal risk of contamination, closed photobioreactors offer better control on culture conditions, such as: CO2 supply, water supply, optimal temperatures, efficient exposure to light, culture density, pH levels, and mixing rates. For a large scale production of biomass, efficient photobioreactors are required. This review paper describes general design considerations pertaining to photobioreactor systems, in order to cultivate microalgae for biomass production. It also discusses the current challenges in designing of photobioreactors for the production of low-cost biomass.

Evidence of sexual transfer of mycobacteria from male to female partners reporting to an IVF clinic.

Female genital tuberculosis (GTB) contributes significantly to infertility in low- and middle-income countries. Dissemination of infection from pulmonary and extrapulmonary sites is the major reason for causation of GTB. Additionally, sexual transmission of GTB from male partners has been reported. We selected 81 couples desiring babies from an in vitro fertilization clinic. We used multiplex-PCR for mycobacterial detection in semen of males, in the endometrium of their female counterparts and in the products of conception (POC) from miscarriage. Data interpretation shows that these pregnancies failed owing to sexual transmission of mycobacteria. We noticed by multiplex PCR that mycobacterial infestation in the female can take place in either endometrium or POC from asymptomatic males harbouring mycobacteria in their semen. Therefore, we propose sexual transfer of mycobacteria to be a probable cause of miscarriage. Thus, we suggest multiplex PCR based screening of semen for all males of the couples attempting successful childbirth.

Mycobacterium tuberculosis FtsZ requires at least one arginine residue at the C-terminal end for polymerization in vitro.

We examined whether C-terminal residues of soluble recombinant FtsZ of Mycobacterium tuberculosis (MtFtsZ) have any role in MtFtsZ polymerization in vitro. MtFtsZ-deltaC1, which lacks C-terminal extreme Arg residue (underlined in the C-terminal extreme stretch of 13 residues, DDDDVDVPPFMRR), but retaining the penultimate Arg residue (DDDDVDVPPFMR), polymerizes like full-length MtFtsZ in vitro. However, MtFtsZ-deltaC2 that lacks both the Arg residues at the C-terminus (DDDDVDVPPFM), neither polymerizes at pH 6.5 nor forms even single- or double-stranded filaments at pH 7.7 in the presence of 10 mM CaCl(2). Neither replacement of the penultimate Arg residue, in the C-terminal Arg deletion mutant DDDDVDVPPFMR, with Lys or His or Ala or Asp (DDDDVDVPPFMK/H/A/D) enabled polymerization. Although MtFtsZ-deltaC2 showed secondary and tertiary structural changes, which might have affected polymerization, GTPase activity of MtFtsZ-deltaC2 was comparable to that of MtFtsZ. These data suggest that MtFtsZ requires an Arg residue as the extreme C-terminal residue for polymerization in vitro. The polypeptide segment containing C-terminal 67 residues, whose coordinates were absent from MtFtsZ crystal structure, was modeled on tubulin and MtFtsZ dimers. Possibilities for the influence of the C-terminal Arg residues on the stability of the dimer and thereby on MtFtsZ polymerization have been discussed.

In vitro polymerization of Mycobacterium leprae FtsZ OR Mycobacterium tuberculosis FtsZ is revived or abolished, respectively, by reciprocal mutation of a single residue.

A single residue that dramatically influences polymerization of principal cell division protein FtsZ of Mycobacterium leprae (MlFtsZ) and Mycobacterium tuberculosis (MtFtsZ) has been identified. Soluble, recombinant MlFtsZ did not show polymerization in vitro, in contrast to MtFtsZ, which polymerised. Mutation of the lone non-conserved residue T172 in the N-terminal domain of MlFtsZ to A172, as it exists in MtFtsZ, showed dramatic polymerization of MlFtsZ-T172A in vitro. Reciprocal mutation of A172 in MtFtsZ to T172, as it exists in MlFtsZ, abolished polymerization of MtFtsZ-A172T in vitro. While T172A mutation enhanced weak GTPase activity of MlFtsZ, reciprocal A172T mutation marginally reduced GTPase activity of MtFtsZ in vitro. These observations demonstrate that the residue at position 172 plays critical role in the polymerization of MlFtsZ and MtFtsZ. A possible evolutionary correlation between the presence of polymerization-adversive or polymerization-favouring residue at position 172 in FtsZ and generation time of the respective bacterium are discussed.

Porous synthetic hectorite clay-alginate composite beads for effective adsorption of methylene blue dye from aqueous solution.

The present study deals with the preparation and characterization of mesoporous synthetic hectorite (MSH) clay which further encapsulated with Na-alginate for the preparation of mesoporous synthetic hectorite-alginate beads (MSH-AB) where Ca2+ act as a cross-linking agent. The detail characterization of MSH and MSH-AB were carried out by various physicochemical techniques. The thermogravimetric analysis study showed better thermal stability results for MSH-AB. The textural properties results of MSH and MSH-AB showed the high surface area 468, 205m2/g, and the pore volume of 0.34, 0.29cm3/g respectively. The applicability of powder MSH and MSH-AB in wet (W) and dry (D) forms were assessed for the removal of cationic dye, methylene blue (MB) by optimizing various batch adsorption parameters. The Langmuir monolayer adsorption capacity obtained for MSH-AB-W showed significant high adsorption efficacy (i.e., 785.45mgMB/g) compared to the MSH-AB-D (357.14mgMB/g) and powder MSH materials (196.00mgMB/g). The adsorption isotherm studies showed that the Langmuir isotherm model was best suitable for MSH, whereas the Freundlich model was utilised to describe the adsorption behavior of organized hydrogel composite beads. The pseudo-second-order kinetics model was observed best for MB sorption onto MSH, whereas pseudo-first order useful to describe the kinetic behavior of MSH-AB. The regeneration experimental results revealed that MSH-AB-W could be recycled more than six cycles with high MB removal efficiency. Furthermore, the adsorption property of the MSH-AB-W was examined for the binary mixture of MB with other dye solutions such as Methyl Red (MR), Methyl Orange (MO), Alizarine Yellow (AY), and Remazol Brilliant Blue (RBB) to evaluate the selective adsorption efficiency. The MSH composite beads were found potentially suitable as an efficient, selective and recyclable adsorbent for the removal of MB from the aqueous solutions.

Myosin-1 inhibition by PClP affects membrane shape, cortical actin distribution and lipid droplet dynamics in early Zebrafish embryos.

Myosin-1 (Myo1) represents a mechanical link between the membrane and actin-cytoskeleton in animal cells. We have studied the effect of Myo1 inhibitor PClP in 1-8 cell Zebrafish embryos. Our results indicate a unique involvement of Myo1 in early development of Zebrafish embryos. Inhibition of Myo1 (by PClP) and Myo2 (by Blebbistatin) lead to arrest in cell division. While Myo1 isoforms appears to be important for both the formation and the maintenance of cleavage furrows, Myo2 is required only for the formation of furrows. We found that the blastodisc of the embryo, which contains a thick actin cortex (~13 µm), is loaded with cortical Myo1. Myo1 appears to be crucial for maintaining the blastodisc morphology and the actin cortex thickness. In addition to cell division and furrow formation, inhibition of Myo1 has a drastic effect on the dynamics and distribution of lipid droplets (LDs) in the blastodisc near the cleavage furrow. All these results above are effects of Myo1 inhibition exclusively; Myo2 inhibition by blebbistatin does not show such phenotypes. Therefore, our results demonstrate a potential role for Myo1 in the maintenance and formation of furrow, blastodisc morphology, cell-division and LD organization within the blastodisc during early embryogenesis.

Enhanced nutrient removal from municipal wastewater assisted by mixotrophic microalgal cultivation using glycerol.

In a present study, nutrient removal from municipal wastewater by Chlorella vulgaris and Nannochloropsis oculata was investigated by using mixotrophic cultivation with glycerol (0 to 5 g/L). Performance parameters were assessed by estimating the removal of total nitrogen, total phosphorus, chemical oxygen demand (COD), biomass growth, chlorophyll content, lipid yield, and fatty acids. With the addition of 2 g/L glycerol, a maximum biomass productivity of 56 mg/L/day was achieved in the mixotrophic culture of C. vulgaris within 12 days. The mixotrophic culture showed a 30-fold increase in biomass productivity compared to the wastewater without any glycerol. However, the highest total nitrogen removal (80.62 %), total phosphate removal (60.72 %), and COD removal (96.3 %) was observed in the N. oculata culture supplemented with 3, 5, and 1 g/L glycerol, respectively. These results suggest that mixotrophic cultivation using glycerol offers great potential in the production of renewable biomass, waste water treatment, and consequent production of high-value microalgal oil. Graphical Abstract Simultaneous biomass production and nutrient removal using microalgae cultivated in wastewater supplemented with glycerol.

Al-intercalated acid activated bentonite beads for the removal of aqueous phosphate.

The separation of fine spent sorbents from treated water after remediation is a major difficulty associated with phosphate wastewater treatment technology. In this study, a novel aluminium-pillared acid activated bentonite powder (Al-ABn) and alginate immobilized aluminium-pillared acid activated bentonite beads (Al-ABn-AB) were synthesized and used for the removal of aqueous phosphate. The phosphate removal behaviour of adsorbents were evaluated by batch experiments as a function of various parameters such as pH, initial concentration, contact time, temperature, adsorbent dose and presence of coexisting ions. The sorption isotherm studies by Langmuir model showed 12.87 and 11.11 mgP/g maximum phosphate uptake capacity for Al-ABn and Al-ABn-AB, respectively. The kinetic studies confirm that the adsorption of phosphate by Al-ABn and Al-ABn-AB follows a pseudo-second-order model. The feasibility of Al-ABn-AB was also assessed in continuous mode in fixed bed column and the loading capacity obtained was 4.55mg/g. The adsorption capacity of Al-ABn-AB beads remained at relatively high even after four regeneration cycles. Furthermore, the applicability of the synthesized adsorbents towards real municipal wastewater confirmed that novel synthesized Al-ABn and Al-ABn-AB are the promising adsorbents for the removal of phosphate from contaminated water.

NDK Interacts with FtsZ and Converts GDP to GTP to Trigger FtsZ Polymerisation--A Novel Role for NDK.

INTRODUCTION: Nucleoside diphosphate kinase (NDK), conserved across bacteria to humans, synthesises NTP from NDP and ATP. The eukaryotic homologue, the NDPK, uses ATP to phosphorylate the tubulin-bound GDP to GTP for tubulin polymerisation. The bacterial cytokinetic protein FtsZ, which is the tubulin homologue, also uses GTP for polymerisation. Therefore, we examined whether NDK can interact with FtsZ to convert FtsZ-bound GDP and/or free GDP to GTP to trigger FtsZ polymerisation. METHODS: Recombinant and native NDK and FtsZ proteins of Mycobacterium smegmatis and Mycobacterium tuberculosis were used as the experimental samples. FtsZ polymersation was monitored using 90° light scattering and FtsZ polymer pelleting assays. The γ32P-GTP synthesised by NDK from GDP and γ32P-ATP was detected using thin layer chromatography and quantitated using phosphorimager. The FtsZ bound 32P-GTP was quantitated using phosphorimager, after UV-crosslinking, followed by SDS-PAGE. The NDK-FtsZ interaction was determined using Ni2+-NTA-pulldown assay and co-immunoprecipitation of the recombinant and native proteins in vitro and ex vivo, respectively. RESULTS: NDK triggered instantaneous polymerisation of GDP-precharged recombinant FtsZ in the presence of ATP, similar to the polymerisation of recombinant FtsZ (not GDP-precharged) upon the direct addition of GTP. Similarly, NDK triggered polymerisation of recombinant FtsZ (not GDP-precharged) in the presence of free GDP and ATP as well. Mutant NDK, partially deficient in GTP synthesis from ATP and GDP, triggered low level of polymerisation of MsFtsZ, but not of MtFtsZ. As characteristic of NDK's NTP substrate non-specificity, it used CTP, TTP, and UTP also to convert GDP to GTP, to trigger FtsZ polymerisation. The NDK of one mycobacterial species could trigger the polymerisation of the FtsZ of another mycobacterial species. Both the recombinant and the native NDK and FtsZ showed interaction with each other in vitro and ex vivo, alluding to the possibility of direct phosphorylation of FtsZ-bound GDP by NDK. CONCLUSION: Irrespective of the bacterial species, NDK interacts with FtsZ in vitro and ex vivo and, through the synthesis of GTP from FtsZ-bound GDP and/or free GDP, and ATP (CTP/TTP/UTP), triggers FtsZ polymerisation. The possible biological context of this novel activity of NDK is presented.

Myosin 1E localizes to actin polymerization sites in lamellipodia, affecting actin dynamics and adhesion formation.

Because the actin network in active lamellipodia is continuously assembling at the edge, moving inward and disassembling, there is a question as to how actin-binding proteins and other components are transported to the leading edge and how nascent adhesions are stabilized. Active transport could play a significant role in these functions but the components involved are unknown. We show here that Myosin 1E (a long tailed Myosin 1 isoform) rapidly moves to the tips of active lamellipodia and to actin-rich early adhesions, unlike Myosin 1G, 1B or 1C (short tailed isoforms). Myosin 1E co-localizes with CARMIL, FHOD1, Arp3 and ß3-integrin in those early adhesions. But these structures precede stable paxillin-rich adhesions. Myosin 1E movement depends upon actin-binding domains and the presence of an SH3 oligomerization domain. Overexpression of a Myosin 1E deletion mutant without the extreme C-terminal interacting (SH3) domain (Myosin 1EΔSH3) increases edge fluctuations and decreases stable adhesion lifetimes. In contrast, overexpression of Myosin 1E full tail domain (TH1+TH2+TH3/SH3) decreases edge fluctuation. In Myosin 1E knockdown cells, and more prominently in cells treated with Myosin 1 inhibitor, cell-matrix adhesions are also short-lived and fail to mature. We suggest that, by moving to actin polymerization sites and early adhesion sites in active lamellipodia, Myosin 1E might play important roles in transporting not only important polymerizing proteins but also proteins involved in adhesion stabilization.

The ftsZ Gene of Mycobacterium smegmatis is expressed Through Multiple Transcripts.

The principal essential bacterial cell division gene ftsZ is differentially expressed through multiple transcripts in diverse genera of bacteria in order to meet cell division requirements in compliance with the physiological niche of the organism under different environmental conditions. We initiated transcriptional analyses of ftsZ gene of the fast growing saprophytic mycobacterium, Mycobacterium smegmatis, as the first step towards understanding the requirements for FtsZ for cell division under different growth phases and stress conditions. Primer extension analyses identified four transcripts, T1, T2, T3, and T4. Transcriptional fusion studies using gfp showed that the respective putative promoter regions, P1, P2, P3, and P4, possessed promoter activity. T1, T2, and T3 were found to originate from the intergenic region between ftsZ and the upstream gene, ftsQ. T4 was initiated from the 3' portion of the open reading frame of ftsQ. RT-PCR analyses indicated co-transcription of ftsQ and ftsZ. The four transcripts were present in the cells at all growth phases and at different levels in the cells exposed to a variety of stress conditions in vitro. T2 and T3 were absent under hypoxia and nutrient-depleted stationary phase conditions, while the levels of T1 and T4 remained unaffected. These studies showed that ftsZ gene expression through multiple transcripts and differential expression of the transcripts at different growth phases and under stress conditions are conserved in M. smegmatis, like in other Actinomycetes.