Co-culture of Saccharomyces cerevisiae (VS3) and Pichia stipitis (NCIM 3498) enhances bioethanol yield from concentrated Prosopis juliflora hydrolysate.

Biphasic acid hydrolysates and enzymatic hydrolysates from carbohydrate-rich Prosopis juliflora, an invasive perennial deciduous shrub of semi-arid regions, were used for bioethanol production. Saccharomyces cerevisiae and Pichia stipitis were used for fermentation of hexoses and pentoses. P. juliflora acid hydrolysate with an initial sugar concentration of 18.70 ± 0.16 g/L was concentrated to 33.59 ± 0.52 g/L by vacuum distillation. The concentrated hydrolysate was pretreated and fermented by mono- and co-culture methods either singly or in combination with enzyme hydrolysate and ethanol yields were compared. Monoculture with S. cerevisiae (VS3) and S. cerevisiae (NCIM3455) yielded maximum ethanol of 36.6 ± 1.83 g/L and 37.1 ± 1.86 g/L with a fermentation efficiency of 83.94 ± 4.20% and 84.20 ± 4.21%, respectively, after 36 h of fermentation. The ethanol yield obtained was 0.428 ± 0.02 g/g substrate and 0.429 ± 0.02 g/g substrate with a productivity of 1.017 ± 0.051 g/L/hand 1.031 ± 0.052 g/L/h, respectively. P. stipitis (NCIM3498) yielded maximum ethanol of 24 g/L with ethanol yield of 0.455 ± 0.02 g/g substrate and a productivity of 1.004 ± 0.050 g/L/h after 24 h of fermentation. With concentrated acid hydrolysate as substrate, S. cerevisiae (VS3) produced ethanol of 8.52 ± 0.43 g/L, whereas S. cerevisiae (NCIM3455) produced 5.96 ± 0.30 g/L of ethanol. P.stipitis (NCIM3498) produced 4.52 ± 0.23 g/L of ethanol by utilizing 14.66 ± 0.87 g/L of sugars. Co-culture with S. cerevisiae (VS3) addition after 18 h of addition of P. stipitis (NCIM3498) to the mixture of concentrated acid hydrolysate and enzyme hydrolysate produced 13.86 ± 0.47 g/L of ethanol with fermentation efficiency, ethanol yield and productivity of 87.54 ± 0.54%, 0.446 ± 2.36 g/g substrate and 0.385 ± 0.014 g/L/h, respectively. Hence, it is concluded that co-culture with S. cerevisiae and P. stipitis is feasible, further scaling up of fermentation of P. juliflora substrate for bioethanol production.

Maize seed endophytic bacteria: dominance of antagonistic, lytic enzyme-producing Bacillus spp.

Beneficial aspects of endophytic microorganisms have motivated researchers to explore plant endophytic world. The present study was aimed to isolate and characterize the seed-borne endophytic bacteria from diverse maize genotypes. Eighty maize seed endophytic bacteria (MSEB), isolated from 30 maize genotypes, were characterized using polyphasic approach. The dendrograms and phylogenetic tree generated on the basis of ARDRA analysis and metabolic profiling of endophytic bacteria revealed genotypic and biochemical diversity among MSEB. The 16S rDNA sequence analysis revealed Bacillus as the most dominant encountered genus affiliated with Phylum Firmicutes. Few isolates belonged to genus Staphylococcus, whereas one isolate was identified as Corynebacterium sp. under Phylum Actinobacteria. Majority of the MSEB isolates exhibited antagonism against phytopathogenic fungi, production of ammonia, and secretion of lytic enzymes; some isolates also exhibited indole acetic acid production, the traits of which can be helpful in endophytic establishment and advantageous to the host plant. Besides, many MSEB exhibited tolerance to salinity (10%), osmotic stress (40% PEG6000), and temperature (60 °C), indicating their possible application under stress conditions. Endophytic nature of the selected MSEB isolates was confirmed by tracking their presence in shoots, leaves, and roots of the host seedlings with the help of biochemical marker (rifampicin resistance). Thus, the MSEB identified in the present study can be explored as potential bioinputs for improving plant growth and productivity under stressed conditions, besides helping in understanding the plant-endophyte interactions.

Selection of suitable mineral acid and its concentration for biphasic dilute acid hydrolysis of the sodium dithionite delignified Prosopis juliflora to hydrolyze maximum holocellulose.

Two grams of delignified substrate at 10% (w/v) level was subjected to biphasic dilute acid hydrolysis using phosphoric acid, hydrochloric acid and sulfuric acid separately at 110 °C for 10 min in phase-I and 121 °C for 15 min in phase-II. Combinations of acid concentrations in two phases were varied for maximum holocellulose hydrolysis with release of fewer inhibitors, to select the suitable acid and its concentration. Among three acids, sulfuric acid in combination of 1 & 2% (v/v) hydrolyzed maximum holocellulose of 25.44±0.44% releasing 0.51±0.02 g/L of phenolics and 0.12±0.002 g/L of furans, respectively. Further, hydrolysis of delignified substrate using selected acid by varying reaction time and temperature hydrolyzed 55.58±1.78% of holocellulose releasing 2.11±0.07 g/L and 1.37±0.03 g/L of phenolics and furans, respectively at conditions of 110 °C for 45 min in phase-I & 121 °C for 60 min in phase-II.

In Vitro Screening for Abiotic Stress Tolerance in Potent Biocontrol and Plant Growth Promoting Strains of Pseudomonas and Bacillus spp.

Plant growth promoting rhizobacteria (PGPR) has been identified as a group of microbes that are used for plant growth enhancement and biocontrol for management of plant diseases. The inconsistency in performance of these bacteria from laboratory to field conditions is compounded due to the prevailing abiotic stresses in the field. Therefore, selection of bacterial strains with tolerance to abiotic stresses would benefit the end-user by successful establishment of the strain for showing desired effects. In this study we attempted to isolate and identify strains of Bacillus and Pseudomonas spp. with stress tolerance and proven ability to inhibit the growth of potential phytopathogenic fungi. Screening of bacterial strains for high temperature (50°C), salinity (7% NaCl), and drought (-1.2 MPa) showed that stress tolerance was pronounced less in Pseudomonas isolates than in Bacillus strains. The reason behind this could be the formation of endospores by Bacillus isolates. Tolerance to drought was high in Pseudomonas strains than the other two stresses. Three strains, P8, P20 and P21 showed both salinity and temperature tolerance. P59 strain possessed promising antagonistic activity and drought tolerance. The magnitude of antagonism shown by Bacillus isolates was also higher when compared to Pseudomonas strains. To conclude, identification of microbial candidate strains with stress tolerance and other added characteristic features would help the end-user obtain the desired beneficial effects.

Selection of the best chemical pretreatment for lignocellulosic substrate Prosopis juliflora.

Pretreatment is a pre-requisite step in bioethanol production from lignocellulosic biomass required to remove lignin and increase the porosity of the substrate for saccharification. In the present study, chemical pretreatment of Prosopis juliflora was performed using alkali (NaOH, KOH, and NH3), reducing agents (Na2S2O4, Na2SO3) and NaClO2 in different concentration ranges at room temperature (30±2 °C) to remove maximum lignin with minimum sugar loss. Further, biphasic acid hydrolysis of the various pretreated substrates was performed at mild temperatures. Considering the amount of holocellulose hydrolyzed and inhibitors released during hydrolysis, best chemical pretreatment was selected. Among all the chemicals investigated, pretreatment with sodium dithionite at concentration of 2% (w/v) removed maximum lignin (80.46±1.35%) with a minimum sugar loss (2.56±0.021%). Subsequent biphasic acid hydrolysis of the sodium dithionite pretreated substrate hydrolyzed 40.09±1.22% of holocellulose and released minimum amount of phenolics (1.04±0.022 g/L) and furans (0.41±0.012 g/L) in the hydrolysate.

Prospective Zinc Solubilising Bacteria for Enhanced Nutrient Uptake and Growth Promotion in Maize (Zea mays L.).

Zinc (Zn) is one of the essential micronutrients required for optimum plant growth. Substantial quantity of applied inorganic zinc in soil is converted into unavailable form. Zinc solubilising bacteria are potential alternates for zinc supplement. Among 10 strains screened for Zn solubilisation, P29, P33, and B40 produced 22.0 mm clear haloes on solid medium amended with ZnCO3. Similarly, P17 and B40 showed 31.0 mm zone in ZnO incorporated medium. P29 and B40 showed significant release of Zn in broth amended with ZnCO3 (17 and 16.8 ppm) and ZnO (18 and 17 ppm), respectively. The pH of the broth was almost acidic in all the cases ranging from 3.9 to 6.1 in ZnCO3 and from 4.1 to 6.4 in ZnO added medium. Short term pot culture experiment with maize revealed that seed bacterization with P29 @ 10 g·kg(-1) significantly enhanced total dry mass (12.96 g) and uptake of N (2.268%), K (2.0%), Mn (60 ppm), and Zn (278.8 ppm).