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.

Enhanced bioethanol production from wheat straw hemicellulose by mutant strains of pentose fermenting organisms Pichia stipitis and Candida shehatae.

The main aim of the present study was to mutate yeast strains, Pichia stipitis NCIM 3498 and Candida shehatae NCIM 3501 and assess the mutant's ability to utilize, ferment wheat straw hemicellulose with enhanced ethanol yield. The organisms were subjected to random mutagenesis using physical (ultraviolet radiation) and chemical (ethidium bromide) mutagens. The mutant and wild strains were used to ferment the hemicellulosic hydrolysates of wheat straw obtained by 2 % dilute sulphuric acid and enzymatic hydrolysis by crude xylanase separately. Among all the mutant strains, PSUV9 and CSEB7 showed enhanced ethanol production (12.15 ± 0.57, 9.55 ± 0.47 g/L and yield 0.450 ± 0.009, 0.440 ± 0.001 g/g) as compared to the wild strains (8.28 ± 0.54, 7.92 ± 0.89 g/L and yield 0.380 ± 0.006 and 0.370 ± 0.002 g/g) in both the hydrolysates. The mutant strains were also checked for their consistency in ethanol production and found stable for 19 cycles in hemicellulosic hydrolysates of wheat straw. A novel element in the present study was introduction of chemical mutagenesis in wild type as well as UV induced mutants. This combination of treatments i.e., UV followed by chemical mutagenesis was practically successful.

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.

Solid state fermentation and production of rifamycin SV using Amycolatopsis mediterranei.

UNLABELLED: Production of Rifamycin SV from cheaper agro-industrial by-products using mutant strain of Amycolatopsis mediterranei OVA5-E7 in solid state fermentation (SSF) was optimized. Among the agro-based substrates used, ragi bran was found suitable for maximizing the yield of Rifamycin SV (1310 mg 100 g(-1) ds). The yield can be further enhanced to 19·7 g Kg(-1) of dry substrate by supplementing the substrate with deoiled cotton cake (10% w/w) using optimized fermentation parameters such as maintaining 80% moisture, pH 7·0, 30°C incubation temperature, inoculum 25% v/w and carrying the solid state fermenting for 9 days. Manipulating these seven specifications, the end product yield achieved in our experimentation was 20 g of Rifamycin SV Kg(-1) ds. Eventually, an overall 5-fold improvement in Rifamycin SV production was achieved. SIGNIFICANCE AND IMPACT OF THE STUDY: Antibiotics such as rifamycin are broad-spectrum antimicrobial drugs used in large-scale worldwide as human medicine towards controlling diseases. Amycolatopsis mediterranei strain which produces this antibiotic was earlier used in submerged fermentation yielded lower amounts of rifamycin. By employing cheaper agro-industrial by-products, we produced upto 20 g rifamycin SV per Kg dry substrate used under optimized solid state fermentation conditions. Keeping in view, the role of rifamycin in meeting the medical demands of world's increasing population; we successfully used an improved strain on cheaper substrates with optimized fermentation parameters and achieved a 5-fold improvement in rifamycin SV production.

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.

Enzymatic hydrolysis of sorghum straw using native cellulase produced by T. reesei NCIM 992 under solid state fermentation using rice straw.

Cellulose is a major constituent of renewable lignocellulosic waste available in large quantities and is considered the most important reservoir of carbon for the production of glucose, for alternative fuel and as a chemical feedstock. Over the past decade, the emphasis has been on the enzymatic hydrolysis of cellulose to glucose and the efficiency of which depends on source of cellulosic substrate, its composition, structure, pretreatment process, and reactor design. In the present study, efforts were made to produce cellulase enzyme using rice straw. The produced enzyme was used for the hydrolysis of selected lignocellulosic substrate, i.e., sorghum straw. When rice straw was used as a substrate for cellulase production under solid state fermentation, the highest enzyme activity obtained was 30.7 FPU/gds, using T. reesei NCIM 992. 25 FPU/g of cellulase was added to differently treated (native, alkali treated, alkali treated followed by 3% acid treated and alkali treated followed by 3 and 5% acid treated) sorghum straw and hydrolysis was carried out at 50 °C for 60 h. 42.5% hydrolysis was obtained after 36 h of incubation. Optimization of enzyme loading, substrate concentration, temperature, time and buffer yielded a maximum of 546.00 ± 0.55 mg/g sugars (54.60 ± 0.44 g/l) with an improved hydrolysis efficiency of 70 ± 0.45%. The enzymatic hydrolyzate can be used for fermentation of ethanol by yeasts.

Development of Dot-ELISA for the detection of human rotavirus antigen and comparison with RNA-PAGE.

AIMS: Development of a simple, specific, rapid and inexpensive Dot-ELISA test for diagnosis of rotaviral antigen in stool samples. METHODS AND RESULTS: Hyperimmune rabbit antisera raised against SA-11 (Simian Agent-11) strain was used as primary antibody. The secondary antibody conjugate used was the goat anti-rabbit IgG alkaline phosphatase, and BCIP/NBT solution was used as substrate. Faecal extracts were diluted 10-fold and used for the detection of rotavirus antigen. RNA-PAGE was performed to compare the specificity and sensitivity of the diagnostic tests. Dot-ELISA positive samples were further confirmed by Western blot analysis. CONCLUSIONS: This Dot-ELISA test could be used as an alternative method for diagnosing rotaviral samples in the field. SIGNIFICANCE AND IMPACT OF THE STUDY: The Dot-ELISA test is simple, specific, rapid and cost effective. It is suitable for identifying a large number of samples obtained from epidemiological studies and hence, reducing the death rate of rotavirus-infected patients.

Effect of uracil on rifamycin SV production by Amycolatopsis mediterranei MV35R.

The effect of different organic nitrogen compounds on the production of rifamycin SV by Amycolatopsis mediterranei MV35R and their optimum concentrations have been described. Results obtained indicate that rifamycin SV production increased from 4020 mg l-1 to 4575 mg l-1 when organic nitrogen compound uracil was added at 0.2% (w/v) concentration to the fermentation medium by A. mediterranei MV35R. The rifamycin SV yield was enhanced by 505 mg l-1 using uracil (2 g l-1) when compared with barbital.