Mutagenesis enhances gellan gum production by a novel Sphingomonas spp.: upstream optimization, kinetic modeling, and structural and physico-functional evaluation.

Gellan gum (GG) has gained tremendous attention owing to its diversified applications. However, its high production and hence market cost are still a bottleneck in its widespread utilization. In the present study, high GG producing mutant of Sphingomonas spp. was developed by random mutagenesis using ethyl methylsulphonate (EMS) for industrial fermentation and identified as Sphingomonas trueperi after 16S rRNA and matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS) analysis. The fermentation conditions such as pH, temperature, and inoculum ratio were optimized by one factor at a time (OFAT) followed by screening of medium components by the Plackett-Burman statistical design. The most critical nutrients were further optimized by response surface methodology for maximizing GG production. The effect of dissolved oxygen tension in bioreactor on cell growth, substrate consumption, GG production, and batch productivity was elucidated. The highest GG titer (23 ± 2.4 g/L) was attained in optimized medium at 10% inoculum (6.45 ± 0.5 log cfu/mL) under controlled fermentation conditions of pH (7), temperature (30 °C), agitation (300-600 rpm), and aeration (0.5-2.0 SLPM) at 22 ± 2% dissolved oxygen tension in a 10-L bioreactor. Kinetic modeling of optimized batch process revealed that logistic growth model could best explain biomass accumulation, while GG formation and substrate consumption were best explained by Luedeking-Piret and exponential decay model, respectively. Structural and physico-functional features of GG produced by mutant Sphingomonas spp. were characterized by HPLC, FTIR, NMR, DSC, TGA, GPC, SEM, and rheological analysis. The higher productivity (0.51 g/L/h) under optimized fermentation conditions suggests potential consideration of mutant and process for commercial utilization.

A supercritical fluid co-extract of turmeric powder and dried coconut shreds shows neuroprotection against AlCl3-induced Alzheimer's disease in rats through nose to brain delivery.

This study was aimed at investigating the neuroprotective potential of a co-extract obtained by supercritical fluid extraction (SFE) of turmeric powder and dried coconut shreds against aluminium chloride (AlCl3)-induced Alzheimer's disease (AD) in male Wistar rats. Fifty animals were allocated to five groups, which received saline (vehicle control, group 1), a combination of saline and aluminium chloride (AlCl3) (disease control, group 2), coconut oil (COO) (SFE extracted, treatment group 3), turmeric oleoresin (Cur) (SFE extracted, treatment group 4) and SFE co-extract of turmeric powder and coconut shreds (CurCOO) (treatment group 5). Animals were subjected to behavioural evaluation. In addition, the hippocampal section of the brain from all groups was subjected to biochemical, molecular and histopathological evaluations. The results showed CurCOO administered intranasally improved cognitive abilities, reversed histological alterations in the brain, reduced hippocampus inflammation studied through proinflammatory cytokine markers like TNF-α and IL-6 as compared to the disease control group. The impact of CurCOO on preventive neurodegeneration was also observed through a reduction in protein transcription factor NF-kB in the treated group 5 as compared to a disease control group. The effect of intranasal delivery of CurCOO on the neurons responsible for memory consolidation was evident from low acetylcholinesterase (AChE) enzyme activity in the treated groups with respect to AlCl3 induced group. Summarily, the results demonstrated intranasal delivery of CurCOO to show better efficacy than Cur and COO in preventing neurodegeneration associated with AlCl3 induced Alzheimer's disease.

Investigation into the chemical modification of α-amylase using octenyl succinic anhydride: enzyme characterisation and stability studies.

The present study describes the chemical modification of α-amylase using succinic anhydride (SA), phthalic anhydride (PA) and a novel modifier viz. 2-octenyl succinic anhydride (2-OSA). SA-, PA- and 2-OSA-α-amylases displayed a 50%, 91% and 46% increase in stability at pH 9, respectively; as compared to unmodified α-amylase. PA-α-amylase showed a significant increase in Ea and ΔHa#, and a concomitant decrease in ΔSa#. The modified α-amylases exhibited improved thermostability as reflected by significant reductions in Kd and ΔSd#, and increments in t1/2, D-, Ed, ΔHd# and ΔGd# values. The modified α-amylases displayed variable stabilities in the presence of different surfactants, inhibitors, metal ions and organic solvents. Interestingly, the chemical modification was found to confer resistance against inactivation by Hg2+ on α-amylase. The conformational changes in modified α-amylases were investigated using intrinsic tryptophan fluorescence, ANS (extrinsic) tryptophan fluorescence, and dynamic fluorescence quenching. Both intrinsic and extrinsic tryptophan fluorescence spectra showed increased fluorescence intensity for the modified α-amylases. Chemical modification was found to induce a certain degree of structural rigidity to α-amylase, as shown by dynamic fluorescence quenching. Analysis of the CD spectra by the K2d method using the DichroWeb online tool indicated evident changes in the α-helix, ß-sheet and random coil fractions of the α-amylase secondary structure, following chemical modification using anhydrides. PA-α-amylase exhibited the highest productivity in terms of hydrolysis of starch at 60 °C over a period of 5 h indicating potential in varied biotechnological applications.

Anti-Angiogenic Effect of Cantharellus cibarius Extracts, its Correlation with Lipoxygenase Inhibition, and Role of the Bioactives Therein.

Angiogenesis is a complex physiological process that cannot be treated with single agent therapy. Several edible fungi have been known to encompass bioactive compounds, and are promising sources of multi-component drugs. One such widely consumed edible fungi is Cantharellus cibarius, which has been explored for its biological activities. The present study focused on assessing the anti-angiogenic activity of petroleum ether and ethanol extracts of C. cibarius using chick chorioallantoic membrane (CAM) assay. Both the extracts showed a dose-dependent response which was compared with the anti-angiogenic activity of the positive controls silibinin, and lenalidomide. The extracts were also studied for their lipoxygenase (LOX) inhibitory potential and compared to ascorbic acid as the positive control. The IC50 values of the petroleum ether extract, ethanol extract, and ascorbic acid for LOX inhibition assay were 135.4, 113.1, and 41.5 µg/mL, respectively. Although both the extracts showed similar responses in CAM assay, ethanol extract proved to be more potent in LOX inhibition assay. Finally, the extracts were investigated for their chemical composition using GC-MS. A correlation between LOX inhibition and anti-angiogenic potential was established at the molecular level. A meticulous literature search was carried out to correlate the biochemical composition of the extracts to their anti-angiogenic activity.

Enzymatic response of Moina macrocopa to different sized zinc oxide particles: An aquatic metal toxicology study.

Zinc oxide particles (ZnOPs) of both nanometer and sub-micron sizes are important components of high demand consumer products such as sunscreen, paint, textile, food packaging, and agriculture. Their ultimate discharge in the aquatic ecosystem is nearly unavoidable. For sustainable use of ZnOPs, there is an urgent need to assess its ecotoxicity using ecological indicator organisms. Moina macrocopa, an important component of the aquatic ecosystem is one such less explored indicator organism. In the present investigation, ZnOPs of two different sizes (250 ± 20 and 500 ± 50 nm) were selected for risk assessment as most of the previous reports were based on the use of 10-100 nm ZnOPs. ZnOPs of 500 nm were more lethal than that of 250 nm size, with respective LC50 of 0.0092 ± 0.0012 and 0.0337 ± 0.0133 mg/L against M. macrocopa after 48 h of exposure. We further used a sublethal concentration of 500 nm (0.00336 mg/L) and 250 nm (0.00092 mg/L) ZnOPs followed by measurement of enzymatic biomarkers of toxicity (acetylcholinesterase, digestive enzymes, antioxidant enzymes). A size-dependent variation in enzymatic response to 250 and 500 nm ZnOPs was seen. Exposure to ZnOPs inhibited acetylcholinesterase and digestive enzymes (trypsin, amylase), and elevated antioxidant enzymes (catalase, glutathione S-transferase) levels. The exposure also decreased the superoxide dismutase activity and increased that of ß-galactosidase. Microscopic investigation revealed the accumulation of ZnOPs in the digestive tract of M. macrocopa that possibly disrupts enzyme activities. The present study will contribute to establishing regulatory policy on the maximum permissible limit of ZnOPs in different water bodies.

Enzymatic synthesis of fatty acid esters of trehalose: Process optimization, characterization of the esters and evaluation of their bioactivities.

One-pot synthesis of caprylic, lauric and palmitic acid mono- and diesters of trehalose was catalyzed by the lipase Fermase CALB™ 10000. An optimized molar conversion of 35% of trehalose to its palmitate esters was obtained in acetone at 60 °C with a trehalose:palmitic acid molar ratio of 1:5 in 4 h. Trehalose fatty acid esters (THFAE) were purified by column chromatography and characterized using TLC, HPTLC, HR-MS, ATR-FTIR, and differential scanning calorimetry. THFAE were studied for their antimicrobial potential against four bacterial, and two fungal species. Trehalose monolaurate and trehalose dicaprylate demonstrated MIC of 0.45 mM and 16 mM against Pseudomonas aeruginosa and Escherichia coli, respectively. Trehalose monocaprylate showed the highest inhibition of biofilm forming property against Staphylococcus aureus (86.25%) at 99.2 mM and trehalose dipalmitate had lowest IC50 of 13.23 mM. Furthermore, their anti-inflammatory property was studied in vitro using 15-LOX inhibition assay and human red blood cell membrane stabilization assay. In the confirmatory in vivo tests using carrageenan-induced rat paw edema assay, inflammation in disease control group reached up to 63% as against 32% and 20% for trehalose dilaurate and diclofenac treated groups, respectively. THFAE can hence find potential applications in pharmaceuticals, functional foods, and nutraceuticals.

Cross-linked enzyme aggregates of arylamidase from Cupriavidus oxalaticus ICTDB921: process optimization, characterization, and application for mitigation of acrylamide in industrial wastewater.

Acrylamidase produced by Cupriavidus oxalaticus ICTDB921 was recovered directly from the fermentation broth by ammonium sulfate (40-50%) precipitation and then stabilized by cross-linking with glutaraldehyde. The optimum conditions for the preparation of cross-linked enzyme aggregates of acrylamidase (acrylamidase-CLEAs) were using 60 mM glutaraldehyde for 10 min at 35 °C and initial broth pH of 7.0. Acrylamidase-CLEAs were characterized by SDS-PAGE, FTIR, particle size analyzer and SEM. Cross-linking shifted the optimal temperature and pH from 70 to 50 °C and 5-7 to 6-8, respectively. It also altered the secondary structure fractions, pH and thermal stability along with the kinetic constants, Km and Vmax, respectively. A complete degradation of acrylamide ~ 1.75 g/L in industrial wastewater was achieved after 60 min in a batch process under optimum operating conditions, and the kinetics was best represented by Edward model (R2 = 0.70). Acrylamidase-CLEAs retained ~ 40% of its initial activity after three cycles for both pure acrylamide and industrial wastewater, and were stable for 15 days at 4 °C, retaining ~ 25% of its original activity.

Enhancement of loading and oral bioavailability of curcumin loaded self-microemulsifying lipid carriers using Curcuma oleoresins.

The therapeutic applications of curcumin, a phenolic compound extracted from Curcuma species, is limited due to poor bioavailability. To enhance the bioavailability, self-microemulsifying drug delivery systems (SMEDDS) with curcumin were prepared. Ethyl oleate, Tween 80, and Transcutol® P with surfactant: co-surfactant ratio of 2:1 w/w was selected based on the solubility and pseudo-ternary phase diagrams. The optimized formulation (S-Eo3) was evaluated for use of spice oleoresins as curcumin bioenhancers. The oleophilic phase of curcumin containing SMEDDS formulations was then successfully modified by using bioactive oleoresins extracted from two Curcuma species, viz. C. longa (S-CL1) and C. aromatica (S-CA1). The curcumin content in S-Eo3, S-CL1, and S-CA1 were 69.6 ± 0.23, 82.4 ± 0.62, and 88.8 ± 0.46 mg/g, respectively. Thus, by the partial modification of oleophilic phase of SMEDDS with spice oleoresin (acting as bioenhancer) resulted in ∼88 k improvement of curcumin aqueous solubility. The pharmacokinetic study in male Wistar rats showed that the relative bioavailability of curcumin in S-CL1 and S-CA1 were 26- and 29-fold vis-à-vis 22-fold in S-Eo3 compared to curcumin suspension. All the SMEDDS formulations were stable for three months as established by ICH guidelines.

Evaluation and application of prebiotic and probiotic ingredients for development of ready to drink tea beverage.

Ready-to-drink (RTD) ice tea is a ready prepared tea, produced from green and black tea originating from same plant Camellia sinensis. The objective of this study was to determine the effect of prebiotics [galacto-oligosaccharide (GOS), fructo-oligosaccharide (FOS), and inulin] or synbiotic ingredients (GOS, FOS, inulin, and Lactobacillus acidophilus) on the sensory properties and consumer acceptability of RTD. The quality of green tea extract (GTE) and black tea extract (BTE) was improved with pretreatment of cellulase and pectinase enzymes. The combined enzymatic extraction amplified total extractives up to 76% in GTE and 72% in BTE. Total polyphenol was found to be enhanced to 24% in GTE and 19% in BTE. GTE was further selected for development of RTD in two different formats; synbiotic RTD and prebiotic RTD premix and analyzed for sensory attributes (colour, aroma, taste, and acceptability). Synbiotic RTD was also evaluated for stability over a period of 28 days at 4 °C. Synbiotic RTD developed an unpleasant flavor and aroma during the shelf life. Premix format of RTD developed using spray drying was reconstituted and found to be functionally and sensorially acceptable.

Genetic variation in dihydropyrimidine dehydrogenase (DPYD) gene in a healthy adult Indian population.

BACKGROUND: Dihydropyrimidine dehydrogenase (DPD) encoded by DPYD gene is the major enzyme involved in metabolism of 5-flurouracil (5-FU), a pyrimidine analogue used in cancer chemotherapy. Although very effective as a cancer therapeutic drug, if not rapidly metabolized, 5-FU may prove lethal. Single nucleotide variants (SNVs) within DPYD that modulate DPD enzyme activity contribute to 5-FU toxicity. STUDY: This study looked for DPYD SNVs common in the Indian population that might be associated with variable DPD activity and drug toxicity. To achieve this, sequencing analysis was performed of all 23 exons and flanking intronic regions of the DPYD gene in a cohort of 50 healthy adult Indians. This study detected 22 SNVs including intronic, synonymous and non-synonymous changes in the DPYD gene, of which six have not been documented before. Allelic frequency was calculated for the observed variants and linkage disequilibrium (LD) analysis was performed on variants with frequency ≥0.1 to identify haplotypes. CONCLUSIONS: This study provides a brief overview of the genetic polymorphism in DPYD in Indians and emphasizes the need for a large scale extensive study to establish markers associated with the frequently observed variable drug metabolism.

Enhanced stability of alcohol dehydrogenase by non-covalent interaction with polysaccharides.

Non-covalent interaction of alcohol dehydrogenase with polysaccharides was studied using three neutral and three anionic polysaccharides. The process of interaction of alcohol dehydrogenase with gum Arabic was optimized with respect to the ratio of enzyme to gum Arabic, pH, and molarity of buffer. Alcohol dehydrogenase-gum Arabic complex formed under optimized conditions showed 93% retention of original activity with enhanced thermal and pH stability. Lower inactivation rate constant of alcohol dehydrogenase-gum Arabic complex within the temperature range of 45 to 60 °C implied its better stability. Half-life of alcohol dehydrogenase-gum Arabic complex was higher than that of free alcohol dehydrogenase. A slight increment was observed in kinetic constants (K(m) and V(max)) of gum Arabic-complexed alcohol dehydrogenase which may be due to interference by gum Arabic for the binding of substrate to the enzyme. Helix to turn conversion was observed in complexed alcohol dehydrogenase as compared to free alcohol dehydrogenase which may be responsible for observed stability enhancement.

Degradation of colour in beetroot (Beta vulgaris L.): a kinetics study.

The kinetics of colour (measured as Hunter 'a/b' value) degradation in beetroot puree (Beta vulgaris L.) was studied over a temperature range of 50-120 °C (isothermal process), and also during normal open pan cooking, pressure-cooking and a newly developed and patented fuel-efficient 'EcoCooker' (non-isothermal heating process). The degradation of visual colour as measured as Hunter 'a/b' value was found to follow a first order kinetics, where the rate constant increased with an increase in the temperature. The temperature dependence of degradation was adequately modeled by Arrhenius equation. A mathematical model has been developed using the kinetic parameters obtained from the isothermal experiments to predict the losses of color in the non-isothermal heating/processing method based on the time-temperature data for each of the methods. The results obtained indicate a colour degradation of similar magnitude in all the three modes of cooking used in the study.

Inclusion of konjac glucomannan in pea protein hydrogels improved the rheological and in vitro release properties of the composite hydrogels.

In this study, a composite hydrogel consisting of pea protein and konjac glucomannan (KG) was fabricated using three approaches, namely neutral, salt-set, and alkaline gelation. Hydrogels made from pea protein were brittle and weak. The addition of KG improved the elasticity and water holding capacity of the pea protein hydrogels. Concomitantly, a decrease in syneresis rate and swelling of the composite hydrogels was observed. The alkaline-set hydrogels exhibited the highest resilience to strain. Thixotropicity was found to be less pronounced for salt-set hydrogels. Sulphate had a greater positive effect on the structural recovery and negative effect on hysteresis area than chloride due to the greater salting-out effect of the sulphates. The addition of KG facilitated the formation of an interconnected structure with limited mobility of biopolymer chains. A sharp increase in G' and G" during the temperature ramp indicated the predominance of hydrophobic interactions towards the aggregation of biopolymers. The infrared spectra of the hydrogels revealed a change in secondary structure of proteins on addition of KG. A controlled in vitro release of riboflavin was observed in neutral and salt-set hydrogels. The alkaline-set hydrogels exhibited a prolonged gastric retention time, thereby establishing in vitro antacid activity in the gastric environment.

Composite hydrogels fabricated from konjac glucomannan and gellan gum: Rheological characterization and their potential application in sustainable agriculture.

In this study, konjac glucomannan (KG) was incorporated in high acyl gellan (HAG) and low acyl gellan (LAG) hydrogels in different ratios. The addition of KG increased pseudoplasticity and thermal hysteresis values of the hydrogels. Improvement in elasticity and water holding capacity (WHC) was observed in KG-LAG hydrogels. The highest WHC (98.5 %) was observed for 1K1H (KG:HAG = 1:1) and 3K7L (KG:LAG = 3:7) hydrogels. The crystallinity of the composite hydrogels was lower than hydrogels prepared from individual biopolymers. The hydrogels exhibited a rough surface with minute pores in the cross-section, due to the aggregation of glucomannan on the gellan network in the composite hydrogels. While HAG and 1K1H hydrogels exhibited greater swelling at low pH (3.0), LAG and 3K7L exhibited greater swelling at high pH (11.0). At pH 7.0, the hydrogels exhibited swelling indices >300 %. Incorporation of 1K1H hydrogel at 10 % (w/w) in sandy loamy soil under semi-arid conditions increased the germination of fenugreek microgreens from 60 % to 80 % on the 15th day. Furthermore, the moisture evaporation rate of the soil reduced from 35 % to <15 %, positively impacting the physicochemical properties of the microgreens. The composite hydrogels were successful in achieving a controlled release of phosphate fertilizer.

Mixed Culture Cultivation in Microbial Bioprocesses.

Mixed culture cultivation is well renowned for industrial applications due to its technological and economic benefits in bioprocess, food processing, and pharmaceutical industries. A mixed consortium encompasses to achieve growth in unsterile conditions, robustness to environmental stresses, perform difficult functions, show better substrate utilization, and increase productivity. Hence, mixed cultures are being valorized currently and has also augmented our understanding of microbial activities in communities. This chapter covers a wide range of discussion on recent improvements in mixed culture cultivation for microbial bioprocessing and multifarious applications in different areas. The history of microbial culture, microbial metabolism in mixed culture, biosynthetic pathway studies, isolation and identification of strains, along with the types of microbial interactions involved during their production and propagation, are meticulously detailed in the current chapter. Besides, parameters for evaluating mixed culture performance, large-scale production, and challenges associated with it are also discussed vividly. Microbial community, characteristics of single and mixed culture fermentation, and microbe-microbe interactions in mixed cultures have been summarized comprehensively. Lastly, various challenges and opportunities in the area of microbial mixed culture that are obligatory to improve the current knowledge of microbial bioprocesses are projected.

In silico guided pre-treatment of sugarcane juice with natural inhibitors of polyphenol oxidase (PPO) is a new and effective strategy for development of spray-dried formulation.

Sugarcane juice is a popular beverage and is also processed to produce sugar. The polyphenol oxidase (PPO) in sugarcane juice causes enzymatic browning and makes the process of sugar production complex and cumbersome. Storage of sugarcane juice is also hampered by the high sugar content and rapid microbial fermentation. The present research assessed the potential of lemon juice (LJ) and ginger extract (GE) as natural inhibitors of PPO. Enzyme kinetics and the mechanism of inhibition of LJ and GE were studied. Primary investigation was carried out using molecular docking approach to assess the inhibitory potential of LJ and GE and to determine the nature of interaction between the enzyme and inhibitors. Extracts were used as inhibitors and studies revealed that both reduced the PPO activity. Subsequently, pure bioactive inhibitors such as ascorbic acid, citric acid, and 6-shogaol present in these natural extracts were used to study the mode of inhibition of PPO. Citric acid decreased PPO activity by lowering pH, while ascorbic acid was found to be a competitive inhibitor of PPO with a Ki of 75.69 µM. The proportion of LJ and GE required in sugarcane juice was optimized on the basis of browning index and sensory acceptance. Further, the sugarcane cane juice after inhibition of PPO under optimized conditions was spray dried and evaluated for reconstitution properties. The product formulated in the present study is a new and effective approach to address quality-compromising issues associated with long-term storage of cane juice.

A universal measure of chaotropicity and kosmotropicity.

Diverse parameters, including chaotropicity, can limit the function of cellular systems and thereby determine the extent of Earth's biosphere. Whereas parameters such as temperature, hydrophobicity, pressure, pH, Hofmeister effects, and water activity can be quantified via standard scales of measurement, the chao-/kosmotropic activities of environmentally ubiquitous substances have no widely accepted, universal scale. We developed an assay to determine and quantify chao-/kosmotropicity for 97 chemically diverse substances that can be universally applied to all solutes. This scale is numerically continuous for the solutes assayed (from +361 kJ kg(-1) mol(-1) for chaotropes to -659 kJ kg(-1) mol(-1) for kosmotropes) but there are key points that delineate (i) chaotropic from kosmotropic substances (i.e. chaotropes ≥ +4; kosmotropes ≤ -4 kJ kg(-1) mol(-1) ); and (ii) chaotropic solutes that are readily water-soluble (log P < 1.9) from hydrophobic substances that exert their chaotropic activity, by proxy, from within the hydrophobic domains of macromolecular systems (log P > 1.9). Examples of chao-/kosmotropicity values are, for chaotropes: phenol +143, CaCl(2) +92.2, MgCl(2) +54.0, butanol +37.4, guanidine hydrochloride +31.9, urea +16.6, glycerol [> 6.5 M] +6.34, ethanol +5.93, fructose +4.56; for kosmotropes: proline -5.76, sucrose -6.92, dimethylsulphoxide (DMSO) -9.72, mannitol -6.69, trehalose -10.6, NaCl -11.0, glycine -14.2, ammonium sulfate -66.9, polyethylene glycol- (PEG-)1000 -126; and for relatively neutral solutes: methanol, +3.12, ethylene glycol +1.66, glucose +1.19, glycerol [< 5 M] +1.06, maltose -1.43 (kJ kg(-1) mol(-1)). The data obtained correlate with solute interactions with, and structure-function changes in, enzymes and membranes. We discuss the implications for diverse fields including microbial ecology, biotechnology and astrobiology.

Kinetic modeling and scale up of lipoic acid (LA) production from Saccharomyces cerevisiae in a stirred tank bioreactor.

Scale up studies for production of lipoic acid (LA) from Saccharomyces cerevisiae have been reported in this paper for the first time. LA production in batch mode was carried out in a stirred tank bioreactor at varying agitation and aeration with maximum LA production of 512 mg/L obtained at 350 rpm and 25 % dissolved oxygen in batch culture conditions. Thus, LA production increased from 352 mg/L in shake flask to 512 mg/L in batch mode in a 5 L stirred tank bioreactor. Biomass production under these conditions was mathematically explained using logistic equation and data obtained for LA production and substrate utilization were successfully fitted using Luedeking-Piret and Mercier's models. The kinetic studies showed LA production to be growth associated. Further enhancement of LA production was carried out using fed-batch (variable volume) and semi-continuous modes of fermentation. Semi-continuous fermentation with three feeding cycles of sucrose effectively increased the production of LA from 512 to 725 mg/L.

Separation of polyphenols and arecoline from areca nut (Areca catechu L.) by solvent extraction, its antioxidant activity, and identification of polyphenols.

BACKGROUND: Areca nut (Areca catechu L.) or betel nut, a commercial cash crop, is a rich source of polyphenols but also contains toxic alkaloids, mainly arecoline. Separation of these bioactive polyphenols from toxic constituents could propel the safe and beneficial use of betel nut; also it will help arecanut processing industries to produce arecoline-free products. With the aim to develop an effective method for maximum extraction of polyphenols with minimum arecoline, several factors such as nature of the solvent, pH (2-10), substrate concentration (6-14 %) and extraction time (30-150 min) under shaking conditions were evaluated. Qualitative analysis was done using spectrophotometry and high-performance liquid chromatography (HPLC). RESULTS: Maximum extraction of polyphenols (407.47 mg GAE g(-1)), total tannin and its antioxidant activity with minimum arecoline (1.73 mg g(-1) of sample) was achieved by using 80% acetone at pH 4 for 90 min with 10% w/v substrate under shaking conditions. CONCLUSION: Solvent extraction under optimized parameters gave maximum polyphenols with minimum extraction of arecoline, and highest ratio of polyphenols to arecoline. HPLC and liquid chromatography-mass spectrometry results confirmed the presence of catechin and epicatechin in the extract, which suggests its potential as a source of bioactives.

Characterization and in vitro probiotic evaluation of lactic acid bacteria isolated from idli batter.

An Indian traditional fermented food, idli batter, was used as a source for isolation of lactic acid bacteria (LAB). A total of 15 LAB strains were isolated on the basis of their Gram nature and catalase activity. Of these, one lactobacilli strain and one lactococci strain which showed antimicrobial activity were identified using biochemical characterization, sugar utilization and molecular sequencing. The microbes, labeled as IB-1 (Lactobacillus plantarum) and IB-2 (Lactococcus lactis) were tested for their in vitro tolerance to bile salts, resistance to low pH values and acidifying activity. Both the strains showed good viability (IB1- 58.11%; IB2- 60.84%) when exposed to high bile salt concentration (2%) and acidic pH of ≤pH 3.0 (IB1- 88.13%; IB2- 89.85%). Lactic acid (IB1- 181.93 mM; IB2- 154.44 mM), biomass production (IB1- 0.65; IB2- 0.58 g/l) after 54 h as well as qualitative estimation of ß-galactosidase and vitamin B12 production were also studied to check their suitability as an industrially important strain for production of important biomolecules.