Distillery effluent as a potential medium for bacterial cellulose production: A biopolymer of great commercial importance.

In the present study, an attempt was made to utilize the distillery effluent for the production of bacterial cellulose by a novel bacterial species, Gluconacetobacter oboediens. Maximum bacterial cellulose production of 0.85g/100ml was achieved in crude distillery effluent. The production was successfully scaled up to 1.0L size producing 8.1g of bacterial cellulose. Morphological, structural and thermal characterization of purified bacterial cellulose by SEM, FT-IR and TGA analysis showed that it is pure cellulose having good properties. Henceforth, the present study proved a concept that distillery effluent could be utilized for the production of bacterial cellulose, a biopolymer of immense importance, which in turn may be used for producing different value added products.

Shikimic acid, a base compound for the formulation of swine/avian flu drug: statistical optimization, fed-batch and scale up studies along with its application as an antibacterial agent.

The sudden outbreak of swine flu has increased the global demand of shikimic acid which is an industrially interesting compound, as it is used as a key starting material for the synthesis of a neuraminidase inhibitor Tamiflu(®), for the treatment of antiviral infections such as swine flu. Statistical optimization and evaluation of medium components for the production of shikimic acid by Citrobacter freundii is addressed in the present investigation. Plackett-Burman design was applied for the screening of the most significant variables affecting shikimic acid production, where glucose, asparagine, KH2PO4, CaCO3 and agitation rate were the most significant factors. Response surface methodology was also employed to study the interaction among the most significant variables through which shikimic acid production increased to 12.76 g/L. Further, fed-batch studies resulted in the production of 22.32 g/L of shikimic acid. The scalability of the process was also confirmed by running 14 L bioreactor (7.5 L production medium) where 20.12 g/L of shikimic acid was produced. In addition the antibacterial activity of the shikimic acid produced was analysed against four Gram positive and four Gram negative bacteria and it was found to have a greater inhibition effect against the Gram negative bacteria.

Dual substrate strategy to enhance butanol production using high cell inoculum and its efficient recovery by pervaporation.

The present study deals with the development of an efficient ABE fermentation process using mixed substrate strategy for butanol production wherein no acetone was produced. For this, glucose was supplemented in the medium containing glycerol as main substrate which leads to a higher butanol production of 17.75 g/L in 72 h by Clostridium acetobutylicum KF158795. Moreover, the high cell inoculum also resulted in an increased ABE productivity of 0.46 g/L/h. Further, industrial scalability of the process was also successfully validated in a 300 L fermenter. Furthermore, potential of the Polymeric (PolyRMem) and Zeolite (ZeoMem) membranes for separation of butanol from fermentation broth was also studied by testing the pervaporation performance through which the butanol was successfully recovered.

An interactive study of influential parameters for shikimic acid production using statistical approach, scale up and its inhibitory action on different lipases.

Shikimic acid is the promising candidate as a building block for the industrial synthesis of drug Tamiflu used for the treatment of Swine flu. The fermentative production process using microbes present an excellent and even more sustainable alternative to the traditional plants based extraction methods. In the present study, the fermentative production of shikimic acid by Citrobacter freundii GR-21 (KC466031) was optimized by process engineering using a statistical modeling approach and a maximum amount of 16.78 g L(-1) was achieved. The process was also scaled up to 14L bioreactor to validate the production of shikimic acid. Further, the potential of anti-enzymatic nature of purified shikimic acid was evaluated for different lipases wherein, shikimic acid inhibited the hydrolysis of triglycerides by 55-60%. Shikimic acid also profoundly inhibited pancreatic lipase activity by 66%, thus providing another valuable therapeutic aspect for treating diet induced obesity in humans.

Expanding horizons of shikimic acid. Recent progresses in production and its endless frontiers in application and market trends.

Shikimic acid is an industrially important chiral compound used as a key ingredient in formulation of drug Oseltamivir phosphate (Tamiflu) for the treatment of swine/avian flu. The high cost and limited availability of shikimic acid isolated from plants has detained the use of this valuable building block of the drug. It is a versatile compound having many characteristic properties for many synthetic reactions particularly in pharmaceuticals and cosmetic industries. By virtue of being a natural product, the relevant biochemical pathway in microorganisms can be harnessed into fermentation processes to produce shikimic acid. This is an excellent alternative for the sustainable and efficient production of shikimic acid over the tedious and cumbersome process of plant based extraction methods. Various strategies of shikimic acid production are reviewed and an account of comparison of their challenges, promises and restraint is presented. Furthermore, present review attempts to focus on the market trend of shikimic acid due to its high demand with particular emphasis laid on the pandemics of swine flu. This review not only covers the recent advances in shikimic acid production but also highlights the versatile applications and its market scenario. The concluding remarks and its potential as a commercial bulk chemical are discussed in the light of current research.

Fermentative production of shikimic acid: a paradigm shift of production concept from plant route to microbial route.

Different physiological and nutritional parameters affect the fermentative production of shikimic acid. In our study, Citrobacter freundii initially produced 0.62 g/L of shikimic acid in 72 h. However, when process optimization was employed, 5.11 g/L of shikimic acid was produced in the production medium consisting of glucose (5.0 %), asparagine (4.5 %), CaCO3 (2.0 %), at pH 6.0, when inoculated with 6 % inoculum and incubated at 30 ± 1 °C, 200 rpm for 60 h. Preliminary fed-batch studies have resulted in the production of 9.11 g/L of shikimic acid on feeding the production medium by 20 g/L of glucose at 24 h of the fermentation run. Production of similar amount of shikimic acid was observed when the optimized conditions were employed in a 10-L bioreactor as obtained in shake flask conditions. A total of 9.11 g/L of shikimic acid was produced in 60 h. This is approximately 14.69-fold increase in shikimic acid production when compared to the initial un-optimized production conditions. This has also resulted in the reduction of the production time. The present study provides useful information to the industrialists seeking environmentally benign technology for the production of bulk biomolecules through manipulation of various chemical parameters.

Evaluation of corncob hemicellulosic hydrolysate for xylitol production by adapted strain of Candida tropicalis.

A maximum xylose extraction of 21.98 g/L was obtained in hydrolysate with a solid to liquid ratio of 1:8 (w/v) at 1% H(2)SO(4) and treated for 30 min. The optimized and treated corncob hemicellulosic hydrolysate medium supplemented with (g/L) yeast extract 5.0, KH(2)PO(4) 2.0, MgSO(4)·7H(2)O 0.3 and methanol 10 mL whose pH was adjusted to 4.5 acts as production medium. Under this condition; the adapted strain of C. tropicalis resulted in 1.22-fold increase in xylitol yield and 1.70-fold enhancement in volumetric productivity was obtained as compared to parent strain of C. tropicalis. On concentrating the hydrolysate under vacuum using rotavapor proves to be efficient in terms of improved xylitol yield and productivity over microwave assisted concentration using adapted strain of C. tropicalis. The immobilized cells of C. tropicalis resulted in more than 70% efficiency up to third cycle. The xylitol production could be scaled up to 10 L fermentor.

A natural isolate producing shikimic acid: isolation, identification, and culture condition optimization.

Shikimic acid has wide use in pharmaceuticals due to its application in the synthesis of drug Tamiflu used in the treatment of Swine flu. The high cost and limited availability of shikimic acid isolated from plants has impeded the use of this valuable building block of the drug. In this context, fermentation route to produce shikimic acid from renewable resources has become increasingly attractive. The present study was embarked upon isolation of wild-type microorganisms able to produce shikimic acid. Out of the 42 isolates obtained from the soil, isolate GR-21 was selected as the best with initial production of 0.54 g/L shikimic acid and later identified as Citrobacter sp. The process optimization resulted in 14-fold increase in the shikimic acid production, thereby claiming this process to be a sustainable alternative for the production of this important biomolecule. The process was further scaled up to 14 L bioreactor to validate the production of shikimic acid. Further, the product formed is shikimic acid was confirmed by FTIR analysis. The current studies suggest that the selected isolate could be used as a promising agent to fulfill the worldwide demand of shikimic acid.

Efficient production of L-asparaginase from Bacillus licheniformis with low-glutaminase activity: optimization, scale up and acrylamide degradation studies.

L-Asparaginase has potential as an anti-cancer drug and for prevention of acrylamide formation in fried and baked foods. Production of the enzyme by Bacillus licheniformis (RAM-8) was optimized by process engineering using a statistical modeling approach and a maximum yield of 32.26 IU/ml was achieved. The L-asparaginase exhibited glutaminase activity of only 0.8 IU/ml and would therefore be less prone to cause the side effects associated with asparaginase therapy compared to enzyme preparations with higher glutaminase activities. When production was carried out in a 30-L bioreactor, enzyme production reached 29.94 IU/ml in 15 h. The enzyme inhibited poly-acrylamide formation in 10% acrylamide solution and reduced acrylamide formation in fried potatoes by 80%.

Pandemism of swine flu and its prospective drug therapy.

Swine flu is a respiratory disease caused by influenza A H1N1 virus. The current pandemic of swine flu is most probably due to a mutation-more specifically, a re-assortment of four known strains of influenza A virus subtype H1N1. Antigenic variation of influenza viruses while circulating in the population is an important factor leading to difficulties in controlling influenza by vaccination. Due to the global effect of swine flu and its effect on humans, extensive investigations are being undertaken. In this context, Tamiflu is the only available drug used in the prophylaxis of this disease and is made from the compound shikimic acid. Due to the sudden increase in the demand of shikimic acid, its price has increased greatly. Thus, it is necessary to find an alternative approach for the treatment of swine flu. This review presents the overall information of swine flu, beginning from its emergence to the prevention and treatment of the disease, with a major emphasis on the alternative approach (bacterial fermentation process) for the treatment of swine flu. The alternative approach for the treatment of swine flu includes the production of shikimic acid from a fermentation process and it can be produced in large quantities without any time limitations.

Examine growth inhibition pattern and lactic acid production in Streptococcus mutans using different concentrations of xylitol produced from Candida tropicalis by fermentation.

Twenty clinical isolates of Streptococcus sp. were isolated from six clinical samples of dental caries on MSFA. Amongst these isolates, five clinical isolates were identified as S treptococcus mutans on the basis of morphological, biochemical and 16S rDNA sequencing. The isolated strains of S. mutans were exposed to fermented and purified xylitol (0.25-15.0%) and tested for its anti-microbial effects against control medium (Brain Heart Infusion without xylitol) after 12 h. The plate assay was developed using bromocresol green as an indicator dye in order to study the relative growth inhibition pattern of clinical sample at different concentrations of an anti-microbial compound in a single petriplate. The morphology of S. mutans cells in brain heart infusion (BHI) medium containing xylitol resulted in a diffused cell wall as observed using gram staining technique. The minimum inhibitory concentration (MIC) is 0.25% for S. mutans obtained from different clinical samples. The MIC(50) and MIC(90) is 5.0% and 10.0% xylitol respectively of the selected S. mutans being designated as clinical isolate B (6). The zone of inhibition was 72 mm and lactic acid production was 0.010 g/l at 10% xylitol concentration in Brain Heart Infusion Broth.

Production of microbial cellulose by a bacterium isolated from fruit.

This study presents the production of bacterial cellulose (BC) by a bacterium isolated from a rotten fruit and its process optimization. Here, isolation and screening of potent cellulose producers were carried out from different natural sources, viz., soil, rotten fruits, and vegetables and vinegar. A total of 200 bacterial isolates were obtained, which were screened for cellulose production using Hestrin-Schramm medium. A novel and potent cellulose-producing bacterium was newly isolated from a rotten fruit and identified as Gluconacetobacter sp. F6 through 16S ribosomal DNA sequencing and morphological, cultural, and biochemical characteristics. After optimization of culture conditions, including pH, temperature, agitation, carbon/nitrogen sources, and inducers, the BC production was greatly increased from 0.52 to 4.5 g/l (8.65-fold increase). The optimal culture medium contained 1% (w/v) glucose, 1.5% (w/v) yeast extract, 0.5% (w/v) peptone, 0.27% (w/v) disodium hydrogen phosphate, 0.115% (w/v) citric acid, and 0.4% (w/v) ethanol. BC produced was analyzed for the presence of cellulose fibrils by epiflourescent microscopy using Calcofluor white stain and scanning electron microscopy and confirmed by NMR. There are very scanty reports about the optimization of BC production by bacteria isolated from rotten fruits.

Fermentation behavior of osmophilic yeast Candida tropicalis isolated from the nectar of Hibiscus rosa sinensis flowers for xylitol production.

Eighteen yeast species belonging to seven genera were isolated from ten samples of nectar from Hibiscus rosa sinensis and investigated for xylitol production using D-xylose as sole carbon source. Amongst these isolates, no. 10 was selected as the best xylitol producer and identified as Candida tropicalis on the basis of morphological, biochemical and 26S rDNA sequencing. C. tropicalis produced 12.11 gl(-1) of xylitol in presence of 50 gl(-1) of xylose in 72 h at pH 5, 30°C and 200 rpm. The strain of C. tropicalis obtained through xylose enrichment technique has resulted in a yield of 0.5 gg(-1) with a xylitol volumetric productivity of 1.07 gl(-1)h(-1) in the presence of 300 gl(-1) of xylose through batch fermentation. This organism has been reported for the first time from Hibiscus rosa sinensis flowers. Realizing, the importance of this high valued compound, as a sugar substitute, xylose enrichment technique was developed in order to utilize even higher concentrations of xylose as substrate for maximum xylitol production.

Effect of arbuscular mycorrhizal inoculations on seedling growth and biomass productivity of two bamboo species.

A study was conducted to identify suitable arbuscular mycorrhizal (AM) fungi for inoculation of Bambusa bambos and Dendrocalamus strictus at nursery stage for increasing growth and productivity. Twelve AM species, isolated from bamboo and other common trees of Bundelkhand were used for inoculations. In B. bambos, total dry weight and phosphorus (P) uptake were significantly increased by all studied fungi and shoot length was increased by eight AM inoculants. Maximum mycorrhizal dependency (MD) was recorded for Acaulospora scrobiculata (44.2%), followed by Glomus cerebriforme (41.6%) and G. intraradix (41.0%). In D. strictus, all tested AM inoculants significantly increased shoot length, dry shoot weight and P uptake, except Glomus 1. Dry root weight was significantly increased by only two inoculants namely, G. cerebriforme and G. etunicatum. Total dry weight was significantly increased by eight AM fungi. Maximum MD was recorded for G. cerebriforme (62.9%), followed by G. diaphanum (55.0%) and G. etunicatum (51.3%). Thus, the results showed that utilization of effective AM fungi can enhance the productivity of bamboo in the region.

Comparative Efficacy and Safety of Various Anti-Microbials in Patients of Acute Rhinosinusitis at Tertiary-Care Hospital in Uttarakhand (UK).

To compare the efficacy of gatifloxacin, azithromycin with amoxicillin as positive control for the treatment of Acute rhinosinusitis (ARS). To monitor adverse drug reaction profile of amoxicillin, azithromycin and gatifloxacin in patient of ARS. An open randomized trial of comparative efficacy and safety of amoxicillin, azithromycin and gatifloxacin in patients with ARS. Patients were randomized into three groups as under: group 1: patients on oral amoxicillin 500 mg TDS for 10 days; group 2: patients on oral azithromycin 500 mg OD for 5 days; group 3: patients on oral gatifloxacin 400 mg OD for 10 days. Patients were evaluated for signs and symptoms at day 1, day 7 (for group 2), on day 12 (for group 1, 3) as primary end points and 28 days after post therapy as secondary end point. All the three antimicrobial drugs i.e., amoxicillin, azithromycin and gatifloxacin were effective in reducing symptoms of acute sinusitis on visual analogue scale scoring. Azithromycin showed significant improvement radiographically on day 7 (P < 0.01) and on day 35 (P < 0.01). Gatifloxacin elicited very highly significant improvement radiographically on day 40 (P < 0.001) and significant improvement on day 12 (P < 0.01) when compared with amoxicillin. Azithromycin, the ketolide was associated with lesser adverse effects as compared to amoxicillin. All the three antimicrobial drugs i.e., amoxicillin, azithromycin and gatifloxacin were effective in reducing symptoms of acute sinusitis on visual analogue scale scoring. Gatifloxacin was found to be most effective drug both in terms of improvement in signs and symptoms on visual analogue scale and radiographic scoring and was associated with least adverse events in comparison to rest of two drugs under study.

Pigeonpea genomics initiative (PGI): an international effort to improve crop productivity of pigeonpea (Cajanus cajan L.).

Pigeonpea (Cajanus cajan), an important food legume crop in the semi-arid regions of the world and the second most important pulse crop in India, has an average crop productivity of 780 kg/ha. The relatively low crop yields may be attributed to non-availability of improved cultivars, poor crop husbandry and exposure to a number of biotic and abiotic stresses in pigeonpea growing regions. Narrow genetic diversity in cultivated germplasm has further hampered the effective utilization of conventional breeding as well as development and utilization of genomic tools, resulting in pigeonpea being often referred to as an 'orphan crop legume'. To enable genomics-assisted breeding in this crop, the pigeonpea genomics initiative (PGI) was initiated in late 2006 with funding from Indian Council of Agricultural Research under the umbrella of Indo-US agricultural knowledge initiative, which was further expanded with financial support from the US National Science Foundation's Plant Genome Research Program and the Generation Challenge Program. As a result of the PGI, the last 3 years have witnessed significant progress in development of both genetic as well as genomic resources in this crop through effective collaborations and coordination of genomics activities across several institutes and countries. For instance, 25 mapping populations segregating for a number of biotic and abiotic stresses have been developed or are under development. An 11X-genome coverage bacterial artificial chromosome (BAC) library comprising of 69,120 clones have been developed of which 50,000 clones were end sequenced to generate 87,590 BAC-end sequences (BESs). About 10,000 expressed sequence tags (ESTs) from Sanger sequencing and ca. 2 million short ESTs by 454/FLX sequencing have been generated. A variety of molecular markers have been developed from BESs, microsatellite or simple sequence repeat (SSR)-enriched libraries and mining of ESTs and genomic amplicon sequencing. Of about 21,000 SSRs identified, 6,698 SSRs are under analysis along with 670 orthologous genes using a GoldenGate SNP (single nucleotide polymorphism) genotyping platform, with large scale SNP discovery using Solexa, a next generation sequencing technology, is in progress. Similarly a diversity array technology array comprising of ca. 15,000 features has been developed. In addition, >600 unique nucleotide binding site (NBS) domain containing members of the NBS-leucine rich repeat disease resistance homologs were cloned in pigeonpea; 960 BACs containing these sequences were identified by filter hybridization, BES physical maps developed using high information content fingerprinting. To enrich the genomic resources further, sequenced soybean genome is being analyzed to establish the anchor points between pigeonpea and soybean genomes. In addition, Solexa sequencing is being used to explore the feasibility of generating whole genome sequence. In summary, the collaborative efforts of several research groups under the umbrella of PGI are making significant progress in improving molecular tools in pigeonpea and should significantly benefit pigeonpea genetics and breeding. As these efforts come to fruition, and expanded (depending on funding), pigeonpea would move from an 'orphan legume crop' to one where genomics-assisted breeding approaches for a sustainable crop improvement are routine.

Amphotericin B--fat emulsion in treatment of visceral leishmaniasis--a cost effective substitute for liposomal amphotericin B.

In spite of high dosage and prolonged treatment schedule of sodium antimony gluconate, unresponsiveness and relapse ranging from 25 to 40% has been reported. The cure rate of 90 to 92.9% with amphotericin B-fat emulsion is comparable to that with liposomal amphotericin B (80 to 100%). Due to high cost liposomal amphotericin B is beyond the reach of most of the Indian patients, whereas the cost of treatment with amphotericin B-fat emulsion is quite affordable. Further large scale studies with amphotericin B-fat emulsion are required to evaluate its effectiveness in Indian kala-azar patients and to establish its optimal dose.

Expression, purification and characterization of allelic variants of MSP-1(42) from Indian Plasmodium falciparum isolates.

The C-terminal 19 and 42 kDa fragments of Plasmodium falciparum merozoite surface protein 1 (MSP-1) have shown to be protective in animals against lethal parasite challenge. The MSP-1(19) being highly conserved may lack sufficient number of T-cell epitopes in order to elicit a broader response in genetically diverse populations. The inclusion of additional epitopes from the N-terminal MSP-1(42) has shown to enhance the protective efficacy of MSP-1(19) vaccine. In an attempt to examine the strain specific immunogenicity to MSP-1, we have cloned and expressed three diverse allelic variants of MSP-1(42) from Indian P. falciparum isolates in bacteria. Among three alleles, one was extremely rare and not been found before. These purified and refolded recombinant products were recognized by conformation specific monoclonal antibodies and hyper-immune sera. Immunization of mice and rabbits with the purified proteins generated high titer biologically active polyclonal antibodies supporting further development of this vaccine candidate antigen.

Optimization of Lipase Production from Aspergillus terreus by Response Surface Methodology and Its Potential for Synthesis of Partial Glycerides Under Solvent Free Conditions.

Aspergillus terreus produces lipase 7.01 IU/ml in 96 h after optimization by one variable at a time method. Using the significant factors i.e. corn oil (A), sodium nitrate (B), casein (C), agitation rate (D) and incubation period (E) RSM was carried out resulting in 19.65 IU/ml from the combination +1(A), -1(B), -1(C), +1(D) and 0(E). The interactions between sodium nitrate, casein and agitation with corn oil were most significant. Scale up of production from 250 ml shake flask to 30 l bioreactor resulted in increased productivity of 0.52 IU/ml/h as against 0.2 IU/ml/h obtained in shake flasks. This lipase could carryout solvent free synthesis of partial glycerides of oleic acid with 96% efficiency in 12 h.