Development of whey protein beverage incorporating encapsulated probiotic strain Lactiplantibacillus rhamnosus NCDC 347 and its physico-chemical characteristics.

In the present study, encapsulated strain Lactiplantibacillus rhamnosus NCDC 347 was used to prepare a novel whey protein-based beverage. The encapsulation process utilized skimmed milk powder matrix and evaluated strain viability, physico-chemical properties, sensory assessment, and shelf-life stability. Encapsulated L. rhamnosus NCDC 347 within skim milk powder maintained viability at 8.0 log CFU/g, forming spherical microcapsules with 1-12 µm concavities. Probiotic addition to whey protein beverages maintained pH and acidity within desired ranges. Physico-chemical analysis showed protein content of 8.71 ± 0.21 % to 10.05 ± 0.42 %, fat content of 0.56 ± 0.24 % to 0.67 ± 0.13 %, viscosity of 5.14 pa/s, and total soluble solids (TSS) of 14.42 ± 0.31 to 16.16 ± 0.23° Brix. The shelf-life study revealed that the beverage remained stable for up to 90 days with no significant changes (p > 0.05) in sensory analysis. The sensory analysis scored the test sample's acceptability at 7.3 ± 0.41. The protein-rich probiotic drink exhibited favorable sensory qualities. Overall, incorporating encapsulated probiotic strain L. rhamnosus NCDC 347 into whey protein beverages could address daily protein requirements and enhance health.

Development and optimization of an in-house heterologous ELISA for detection of prednisolone drug in enzyme conjugates using spacers.

The introduction of spacers in coating steroid protein complexes and/or enzyme conjugates or immunogens is known to exert an influence on the sensitivity of steroid enzyme immunoassays. We investigated the impact of different homobifunctional spacers, ranging in atomic length from 3 to 10, on the sensitivity and specificity of prednisolone (PSL) enzyme immunoassays. In this study, four homo-bifunctional spacers, namely, carbohydrazide (CH), adipic acid dihydrazide (ADH), ethylene diamine (EDA), and urea (U), were incorporated between PSL and horseradish peroxidase (HRP) for preparing the enzyme conjugate with an aim to improve the sensitivity of the assay without compromising assay specificity. The assays were developed using these enzymes conjugated with antibodies raised against the PSL-21-HS-BSA immunogen. The sensitivity of the PSL assays after insertion of a bridge in the enzyme conjugate was 1.22 ng/mL, 0.59 ng/mL, 0.48 ng/mL, and 0.018 ng/mL with ADH, CH, EDA, and urea as a spacer, respectively. Among the four combinations, the PSL-21-HS-BSA-antibody with PSL-21-HS-U-HRP-enzyme conjugate gave better sensitivity and less cross-reaction. The percent recovery of PSL from the exogenously spiked human serum pools was in the range of 88.32%-102.50%. The intra and inter-assay CV% was< 8.46%. The PSL concentration was estimated in the serum samples of patients on PSL treatment. The serum PSL values obtained by this method correlated well with the commercially available kit (r2 = 0.98). The present study suggests that the nature of the spacer is related to assay sensitivity and not the spacer length.

Germination induced changes in bioactive compounds and nutritional components of millets.

The present study was conducted to compare the nutritional quality and the concentration of bioactive compounds in the flours from dehusked and germinated foxtail, barnyard, and little millets. Germinated millet flours showed significantly higher protein content (11.79-33.49%), total free amino acid content (66-334.87%) and protein solubility (13.83-34%), compared to the dehusked millet flours. The total phenols and flavonoids in the flours from the three germinated millets were significantly higher by about 142.36 and 437.20%, respectively, compared to the flours from the dehusked millets. Mineral content was also found to be higher in the flours from germinated millets in comparison to the flours from dehusked millets. The results of this study showed that the flours from the germinated millets have the potential for their application in development of novel products,because of their enhanced nutritional value. The extracts from the germinated millets have the potential for use as functional ingredients in the development of novel processed fruit beverages.

Hydrolysis of peanut (Arachis hypogea L) protein concentrate by fungal crude protease extract: effect on structural, functional and in-vitro protein digestibility.

Peanut protein concentrates (PPCs) were subjected to hydrolysis by crude protease extract (CPE) obtained from three fungi viz; Rhizopus oligosporus, Trichoderma reesei, and Aspergillus oryzae and the effect on structural, functional and in-vitro protein digestibility (IVPD) properties were studied. Particle size was found significantly (p ≤ 0.05) lower in hydrolyzed samples than un-treated samples. Fourier transform infrared spectroscopy (FTIR) spectrum of hydrolyzed samples displayed intense absorbance peaks in the wavelength ranging from 1500 to 2600 cm-1. Peanut protein concentrates hydrolyzed by CPE from R. oligosporus showed higher surface hydrophobicity (564.18). Total sulfhydryl content was found lower in all the hydrolyzed samples whereas, reverse trend was observed for exposed sulfhydryl content. The structural changes simultaneously affected the functional and IVPD attributes of hydrolyzed PPCs. In comparison to the PPCs hydrolysed using crude extracts from T. reesei and R. oligosporus, PPCs hydrolysed by A, oryzae showed higher solubility, water and oil binding capacity, foaming capacity and foam stability. Higher IVPD values of 86.70% was also found in PPCs hydrolyzed with CPE of A. oryzae. The study established that CPE hydrolysis of PPCs has potential for scale-up studies and may serve as a cost effective alternative to protein hydrolysis with pure enzymes.

Fungal Genomic Resources for Strain Identification and Diversity Analysis of 1900 Fungal Species.

Identification and diversity analysis of fungi is greatly challenging. Though internal transcribed spacer (ITS), region-based DNA fingerprinting works as a "gold standard" for most of the fungal species group, it cannot differentiate between all the groups and cryptic species. Therefore, it is of paramount importance to find an alternative approach for strain differentiation. Availability of whole genome sequence data of nearly 2000 fungal species are a promising solution to such requirement. We present whole genome sequence-based world's largest microsatellite database, FungSatDB having >19M loci obtained from >1900 fungal species/strains using >4000 assemblies across globe. Genotyping efficacy of FungSatDB has been evaluated by both in-silico and in-vitro PCR. By in silico PCR, 66 strains of 8 countries representing four continents were successfully differentiated. Genotyping efficacy was also evaluated by in vitro PCR in four fungal species. This approach overcomes limitation of ITS in species, strain signature, and diversity analysis. It can accelerate fungal genomic research endeavors in agriculture, industrial, and environmental management.

Ethanol production from sweet sorghum bagasse through process optimization using response surface methodology.

In this study, comparative evaluation of acid- and alkali pretreatment of sweet sorghum bagasse (SSB) was carried out for sugar production after enzymatic hydrolysis. Results indicated that enzymatic hydrolysis of alkali-pretreated SSB resulted in higher production of glucose, xylose and arabinose, compared to the other alkali concentrations and also acid-pretreated biomass. Response Surface Methodology (RSM) was, therefore, used to optimize parameters, such as alkali concentration, temperature and time of pretreatment prior to enzymatic hydrolysis to maximize the production of sugars. The independent variables used during RSM included alkali concentration (1.5-4%), pretreatment temperature (125-140 °C) and pretreatment time (10-30 min) were investigated. Process optimization resulted in glucose and xylose concentration of 57.24 and 10.14 g/L, respectively. Subsequently, second stage optimization was conducted using RSM for optimizing parameters for enzymatic hydrolysis, which included substrate concentration (10-15%), incubation time (24-60 h), incubation temperature (40-60 °C) and Celluclast concentration (10-20 IU/g-dwt). Substrate concentration 15%, (w/v) temperature of 60 °C, Celluclast concentration of 20 IU/g-dwt and incubation time of 58 h led to a glucose concentration of 68.58 g/l. Finally, simultaneous saccharification fermentation (SSF) as well as separated hydrolysis and fermentation (SHF) was evaluated using Pichia kudriavzevii HOP-1 for production of ethanol. Significant difference in ethanol concentration was not found using either SSF or SHF; however, ethanol productivity was higher in case of SSF, compared to SHF. This study has established a platform for conducting scale-up studies using the optimized process parameters.

Improvement in shelf life of minimally processed cilantro leaves through integration of kinetin pretreatment and packaging interventions: Studies on microbial population dynamics, biochemical characteristics and flavour retention.

Effect of integrating optimized combination of pretreatment with packaging on shelf life of minimally processed cilantro leaves (MPCL) was appraised through analysis of their sensory attributes, biochemical characteristics, microbial population and flavour profile during storage. Minimally pretreated cilantro leaves pretreated with 50ppm kinetin and packed in 25µ polypropylene bags showed a shelf life of 21days. Optimized combination helped in efficiently maintaining sensory parameters, flavour profile, and retention of antioxidants in MPCL until 21days. Studies conducted on the effect of optimized combination on microbial population and flavour profile revealed that among different microorganisms, pectinolysers had a significant effect on spoilage of MPCL and their population of ⩽3.59logcfu/g was found to be acceptable. Principal component analysis of headspace volatiles revealed that (E)-2-undecenal, (E)-2-hexadecenal, (E)-2-tetradecenal & (E)-2-tetradecen-1-ol in stored samples clustered with fresh samples and therefore, could be considered as freshness indicators for MPCL.

An acidothermophilic functionally active novel GH12 family endoglucanase from Aspergillus niger HO: purification, characterization and molecular interaction studies.

Endoglucanase (EG) from Aspergillus niger HO was sequentially purified through ultrafiltration, ion exchange and size exclusion chromatography to homogeneity, with an overall recovery of 18 %. The purified EG was a monomeric protein with a molecular weight of about 55 kDa. The enzyme was optimally active at pH 3.5 and 70 °C with a half life (t1/2) of 3 h and Km value of 2.5 mg/ml. Metal ions, such as Ca(2+) and Co(2+) helped in enzyme induction, while Hg(2+) and Cu(2+) strongly inhibited the enzyme activity. Peptide mass fingerprinting results revealed that the purified EG is a novel enzyme that belongs to family 12 of glycoside hydrolase (GH12). Molecular docking studies indicated the presence of Glu116 and Glu204 as important determinant residues for the functional interaction with carboxymethylcellulose and showed hydrogen bonding with Asp99, Glu116, Glu204 and hydrophobic interactions with Trp22, Val58, Tyr61, Phe101, Met118, Trp120, Pro129, Ile130, Thr160 and Phe206. Hydrolysis of 2 % CMC with purified acidothermophilic EG at its optimum temperature and pH resulted in complete hydrolysis within 2 h yielding 18 % cellotriose, 72 % cellobiose and 10 % glucose as evident from HPLC analysis. In comparison to most of the EGs reported in literature, EG from A. niger HO exhibited higher thermostability. The acidothermophilic nature of this enzyme makes it potentially useful for industrial applications.

Therapeutic and nutraceutical potential of bioactive compounds extracted from fruit residues.

The growing interest in the substitution of synthetic food antioxidants by natural ones has fostered research in identifying new low-cost antioxidants having commercial potential. Fruits such as mango, banana, and those belonging to the citrus family leave behind a substantial amount of residues in the form of peels, pulp, seeds, and stones. Due to lack of infrastructure to handle a huge quantity of available biomass, lack of processing facilities, and high processing cost, these residues represent a major disposal problem, especially in developing countries. Because of the presence of phenolic compounds, which impart nutraceutical properties to fruit residues, such residues hold tremendous potential in food, pharmaceutical, and cosmetic industries. The biological properties such as anticarcinogenicity, antimutagenicity, antiallergenicity, and antiageing activity have been reported for both natural as well as synthetic antioxidants. Special attention is focused on extraction of bioactive compounds from inexpensive or residual sources. The purpose of this review is to characterize different phenolics present in the fruit residues, discuss the antioxidant potential of such residues and the assays used in determination of antioxidant properties, discuss various methods for efficient extraction of the bioactive compounds, and highlight the importance of fruit residues as potential nutraceutical resources and biopreservatives.

Influence of different solvents in extraction of phenolic compounds from vegetable residues and their evaluation as natural sources of antioxidants.

Dried residues from four different vegetables, viz. pea pod (pp), cauliflower waste (CW), potato peel (PP) and tomato peel (TP) were extracted using four solvents i.e., hexane, chloroform, ethyl acetate and methanol. Among the four solvents, methanolic extracts showed the highest total phenolic content (TPC) for all the four vegetable residues. Methanolic extracts were evaluated for antioxidant activities using diphenylpicryl-hydrazyl (DPPH) and reducing power assay. Tomato peel extract showed highest phenolic content of 21.0 mg GAE/g-dw and 80.8 % DPPH free radical scavenging ability, whereas potato peel extract had a low phenolic content, and it also showed the least antioxidant activity among the residues examined in this study. Total phenolic content and DPPH free radical scavenging activity in pea pods and cauliflower waste were 13.6 mg GAE/g-dw and 72 % and 9.2 mg GAE/g-dw and 70.7 %, respectively. The coefficient of determination (r(2)) for correlation between TPC and reducing power, DPPH and TPC, DPPH and reducing power for all extracts was 0.85, 0.91and 0.87, respectively, suggesting an important role of phenolics in imparting antioxidant ability. Extracts from vegetables residues therefore represent a significant source of phenolic antioxidants for use as nutraceuticals or biopreservatives.

Cellulolytic and xylanolytic enzymes from thermophilic Aspergillus terreus RWY.

Thermophilic Aspergillus terreus RWY produced cellulases and xylanases in optimal concentrations at 45 °C in solid state fermentation process, though enzyme production was also observed at 50 and 55 °C. Filter paper cellulase (FP), endoglucanase (EG), ß-glucosidase (BGL), cellobiohydrolase (CBH), xylanase, ß-xylosidase, α-L-arabinofuranosidase and xylan esterase activities for A. terreus RWY at 45 °C in 72 h were 11.3 ± 0.65, 103 ± 6.4, 122.5 ± 8.7, 10.3 ± 0.66, 872 ± 22.5, 22.1 ± 0.75, 126.4 ± 8.4 and 907 ± 15.5 U (g-ds)(-1) , respectively. Enzyme was optimally active at temperatures and pH ranging between 50-60 °C and 4.0-6.0, respectively. The half life (T1/2 ) of 270 and 240 min at 70 and 75 °C, respectively for the enzyme indicates its stability at higher temperatures. The addition of MnCl2 , CoCl2 , and FeCl3 significantly enhanced cellulase activity. Enzyme demonstrated multiplicity by having seven, one and three isoform(s) for EG, CBH and BGL, respectively. Significant production of functionally active consortium of cellulolytic and xylanolytic enzymes from A. terreus RWY makes it a potential candidate in bioprocessing applications.

Effect of nickel-cobaltite nanoparticles on production and thermostability of cellulases from newly isolated thermotolerant Aspergillus fumigatus NS (class: Eurotiomycetes).

In the present study, effect of nickel-cobaltite (NiCo2O4) nanoparticles (NPs) was investigated on production and thermostability of the cellulase enzyme system using newly isolated thermotolerant Aspergillus fumigatus NS belonging to the class Euratiomycetes. The NiCo2O4 NPs were synthesized via hydrothermal method assisted by post-annealing treatment and characterized through X-ray diffraction (XRD) and transmission electron microscopy (TEM) techniques. In the absence of NPs in the growth medium, filter paper cellulase (FP) activity of 18 IU/gds was achieved after 96 h, whereas 40 % higher FP activity in 72 h was observed with the addition of 1 mM concentration of NPs in the growth medium. Maximum production of endoglucanase (211 IU/gds), ß-glucosidase (301 IU/gds), and xylanase (803 IU/gds) was achieved after 72 h without NPs (control), while in the presence of 1 mM concentration of NPs, endoglucanase, ß-glucosidase, and xylanase activity increased by about 49, 53, and 19.8 %, respectively, after 48 h of incubation, against control, indicating a substantial increase in cellulase productivity with the addition of NiCo2O4 NPs in the growth medium. Crude enzyme was thermally stable for 7 h at 80 °C in presence of NPs, as against 4 h at the same temperature for control samples. Significant increase in the activity and improved thermal stability of cellulases in the presence of the NiCo2O4 NPs holds potential for use of NiCo2O4 NPs during enzyme production as well as hydrolysis. From the standpoint of biofuel production, these results hold enormous significance.

Response surface optimization for enhanced production of cellulases with improved functional characteristics by newly isolated Aspergillus niger HN-2.

Fungi isolated from partially decayed wood log samples showing characteristic diversity for spore colour, colony morphology and arrangement of spores were assessed for cellulolytic enzyme production. Isolates showing a cellulolytic index of ≥2.0 were assayed for filter paper (FP) cellulase and ß-glucosidase (BGL) production. Molecular characterization confirmed the identity of the selected cellulolytic isolate as a strain of Aspergillus niger (A. niger HN-2). Addition of 2 % (w/v) urea enhanced FP and BGL activity by about 20 and 60 %, respectively. Validation studies conducted at parameters (29 °C, pH 5.4, moisture content 72 % and 66 h) optimized through response surface methodology in a solid-state static tray fermentation resulted in FP, BGL, cellobiohydrolase I (CBHI), endoglucanase (EG), xylanase activity and protein content of 25.3 FPU/g ds, 750 IU/g ds, 13.2 IU/g ds, 190 IU/g ds, 2890 IU/g ds and 0.9 mg/ml, respectively. In comparison, A. niger N402 which is a model organism for growth and development studies, produced significantly lower FP, BGL, CBHI, EG, xylanase activity and protein content of 10.0 FPU/g ds, 100 IU/g ds, 2.3 IU/g ds, 50 IU/g ds, 500 IU/g ds and 0.75 mg/ml, respectively under the same process conditions as were used for A. niger HN-2. Process optimization led to nearly 1.8- and 2.2-fold increase in FP and BGL activity, respectively showing promise for cellulase production by A. niger HN-2 at a higher scale of operation. Zymogram analysis revealed two isoforms each for EG and cellobiohydrolase and three isoforms for BGL. Crude cellulase complex produced by A. niger HN-2 exhibited thermostability under acidic conditions showing potential for use in biofuel industry.

Two-stage statistical medium optimization for augmented cellulase production via solid-state fermentation by newly isolated Aspergillus niger HN-1 and application of crude cellulase consortium in hydrolysis of rice straw.

Cellulolytic enzyme production by newly isolated Aspergillus niger HN-1 was statistically optimized using Plackett-Burman and central composite design (CCD). Optimum concentrations of 2, 0.40, 0.01, and 0.60 g L (-1) for KH2PO4, urea, trace elements solution, and CaCl2·2H2O, respectively, were suggested by Design-Expert software. The two-stage optimization process led to a 3- and 2-fold increases in the filter paper cellulase (FP) and ß-glucosidase activities, respectively. FP, ß-glucosidase, endoglucanase, exopolygalaturonase, cellobiohydrolase, xylanase, α-l-arabinofuranosidase, ß-xylosidase, and xylan esterase activities of 36.7 ± 1.54 FPU gds(-1), 252.3 ± 7.4 IU gds(-1), 416.3 ± 22.8 IU gds(-1), 111.2 ± 5.4 IU gds(-1), 8.9 ± 0.50 IU gds(-1), 2593.5 ± 78.9 IU gds(-1), 79.4 ± 4.3 IU gds(-1), 180.8 ± 9.3 IU gds(-1), and 288.7 ± 11.8 IU gds(-1), respectively, were obtained through solid-state fermentation during the validation studies. Hydrolysis of alkali-treated rice straw with crude cellulases resulted in about 84% glucan to glucose, 89% xylan to xylose, and 91% arabinan to arabinose conversions, indicating potential for biomass hydrolysis by the crude cellulase consortium obtained in this study.

Ethanol production from alkali-treated rice straw via simultaneous saccharification and fermentation using newly isolated thermotolerant Pichia kudriavzevii HOP-1.

In this study, simultaneous saccharification and fermentation (SSF) was employed to produce ethanol from 1% sodium hydroxide-treated rice straw in a thermostatically controlled glass reactor using 20 FPU gds⁻¹ cellulase, 50 IU gds⁻¹ ß-glucosidase, 15 IU gds⁻¹ pectinase and a newly isolated thermotolerant Pichia kudriavzevii HOP-1 strain. Scanning electron micrograph images showed that the size of the P. kudriavzevii cells ranged from 2.48 to 6.93 µm in diameter while the shape of the cells varied from oval, ellipsoidal to elongate. Pichia kudriavzevii cells showed extensive pseudohyphae formation after 5 days of growth and could assimilate sugars like glucose, sucrose, galactose, fructose, and mannose but the cells could not assimilate xylose, arabinose, cellobiose, raffinose, or trehalose. In addition, the yeast cells could tolerate up to 40% glucose and 5% NaCl concentrations but their growth was inhibited at 1% acetic acid and 0.01% cyclohexamide concentrations. Pichia kudriavzevii produced about 35 and 200% more ethanol than the conventional Saccharomyces cerevisiae cells at 40 and 45°C, respectively. About 94% glucan in alkali-treated rice straw was converted to glucose through enzymatic hydrolysis within 36 h. Ethanol concentration of 24.25 g l⁻¹ corresponding to 82% theoretical yield on glucan basis and ethanol productivity of 1.10 g l⁻¹ h⁻¹ achieved using P. kudriavzevii during SSF hold promise for scale-up studies. An insignificant amount of glycerol and no xylitol was produced during SSF. To the best of our knowledge, this is the first study reporting ethanol production from any lignocellulosic biomass using P. kudriavzevii.

Enhanced ethanol production from sugarcane juice by galactose adaptation of a newly isolated thermotolerant strain of Pichia kudriavzevii.

The thermotolerant yeast strain isolated from sugarcane juice through enrichment technique was identified as a strain of Pichiakudriavzevii (Issatchenkiaorientalis) through molecular characterization. The P. kudriavzevii cells adapted to galactose medium produced about 30% more ethanol from sugarcane juice than the non-adapted cells. The recycled cells could be used for four successive cycles without a significant drop in ethanol production. Fermentation in a laboratory fermenter with galactose adapted P. kudriavzevii cells at 40°C resulted in an ethanol concentration and productivity of 71.9 g L(-1) and 4.0 g L(-1)h(-1), respectively from sugarcane juice composed of about 14% (w/v) sucrose, 2% (w/v) glucose and 1% (w/v) fructose. In addition to ethanol, 3.30 g L(-1) arabitol and 4.19 g L(-1) glycerol were also produced, whereas sorbitol and xylitol were not formed during fermentation. Use of galactose adapted P. kudriavzevii cells for ethanol production from sugarcane juice holds potential for scale-up studies.

Ethanol production from banana peels using statistically optimized simultaneous saccharification and fermentation process.

Dried and ground banana peel biomass (BP) after hydrothermal sterilization pretreatment was used for ethanol production using simultaneous saccharification and fermentation (SSF). Central composite design (CCD) was used to optimize concentrations of cellulase and pectinase, temperature and time for ethanol production from BP using SSF. Analysis of variance showed a high coefficient of determination (R(2)) value of 0.92 for ethanol production. On the basis of model graphs and numerical optimization, the validation was done in a laboratory batch fermenter with cellulase, pectinase, temperature and time of nine cellulase filter paper unit/gram cellulose (FPU/g-cellulose), 72 international units/gram pectin (IU/g-pectin), 37 °C and 15 h, respectively. The experiment using optimized parameters in batch fermenter not only resulted in higher ethanol concentration than the one predicted by the model equation, but also saved fermentation time. This study demonstrated that both hydrothermal pretreatment and SSF could be successfully carried out in a single vessel, and use of optimized process parameters helped achieve significant ethanol productivity, indicating commercial potential for the process. To the best of our knowledge, ethanol concentration and ethanol productivity of 28.2 g/l and 2.3 g/l/h, respectively from banana peels have not been reported to date.

Enhanced ethanol production from Kinnow mandarin (Citrus reticulata) waste via a statistically optimized simultaneous saccharification and fermentation process.

Dried, ground, and hydrothermally pretreated Kinnow mandarin (Citrus reticulata) waste was used to produce ethanol via simultaneous saccharification and fermentation (SSF). Central composite design was used to optimize cellulase and pectinase concentrations, temperature, and time for SSF. The D-limonene concentration determined with high-performance liquid chromatography (HPLC) for fresh, dried, and pretreated biomass was 0.76%, 0.32%, and 0.09% (v/w), respectively. Design Expert software suggested that the first-order effect of all four factors and the second-order effect of cellulase and pectinase concentrations were significant for ethanol production. The validation experiment using 6 FPU gds(-1) cellulase and 60 IU gds(-1) pectinase at 37 °C for 12 h in a laboratory batch fermenter resulted in ethanol concentration and productivity of 42 g L(-1) and 3.50 g L(-1) h(-1), respectively. Experiments using optimized parameters resulted in an ethanol concentration similar to that predicted by the model equation and also helped reduce fermentation time.

Evaluation of glycosyl hydrolases in the secretome of Aspergillus fumigatus and saccharification of alkali-treated rice straw.

A thermotolerant Aspergillus fumigatus strain isolated from composting pile of mixed industrial waste was found to produce a spectrum of cellulase and hemicellulases when cultured on rice straw solidified substrate. The two-dimensional electrophoresis (2DE) resolved the secretome into 57 distinct protein spots. The zymograms developed against 2DE gels identified the presence of three ß-glucosidases and five CBHI/EGI isoforms in the secretome. The peptide mass fingerprinting of 17 protein spots by liquid chromatography mass spectrometry characterized the secretome into different glycosyl hydrolase families. The enzyme cocktail produced by A. fumigatus was capable of efficient hydrolysis of alkali pretreated rice straw (at 7% and 10% w/v) resulting in 95% and 91% saccharification, respectively.

Ethanol production from orange peels: two-stage hydrolysis and fermentation studies using optimized parameters through experimental design.

Orange peels were evaluated as a fermentation feedstock, and process conditions for enhanced ethanol production were determined. Primary hydrolysis of orange peel powder (OPP) was carried out at acid concentrations from 0 to 1.0% (w/v) at 121 degrees C and 15 psi for 15 min. High-performance liquid chromatography analysis of sugars and inhibitory compounds showed a higher production of hydroxymethyfurfural and acetic acid and a decrease in sugar concentration when the acid level was beyond 0.5% (w/v). Secondary hydrolysis of pretreated biomass obtained from primary hydrolysis was carried out at 0.5% (w/v) acid. Response surface methodology using three factors and a two-level central composite design was employed to optimize the effect of pH, temperature, and fermentation time on ethanol production from OPP hydrolysate at the shake flask level. On the basis of results obtained from the optimization experiment and numerical optimization software, a validation study was carried out in a 2 L batch fermenter at pH 5.4 and a temperature of 34 degrees C for 15 h. The hydrolysate obtained from primary and secondary hydrolysis processes was fermented separately employing parameters optimized through RSM. Ethanol yields of 0.25 g/g on a biomass basis (YP/X) and 0.46 g/g on a substrate-consumed basis (YP/S) and a promising volumetric ethanol productivity of 3.37 g/L/h were attained using this process at the fermenter level, which shows promise for further scale-up studies.