Studies on changes in microstructure and proteolysis in cow and soy milk curd during fermentation using lactic cultures for improving protein bioavailability.

Cow milk curd was prepared using 2% v/v of Streptococcus thermophilus DG1 and a mixed culture (0.5:1.5 v/v) of S. thermophilus DG1 and Lactobacillus plantarum and incubating at 37 °C for 16 h. Soy milk curd was prepared using different ratios of lactic cultures as stated earlier and also a mixed culture containing S. thermophilus DG1, L. plantarum and Leuconostoc mesenteroides sub spp. mesenteroides in the ratio 1:1:1 v/v along with beet pulp (2% w/w) and incubating at 37 °C for 18 h. This improved functional and probiotic properties of curd. Structural changes in curd samples during fermentation were observed by Scanning Electron Microscope (SEM). Soy milk curd showed loosened structure. The degradation of proteins into peptides and amino acids were evaluated by SDS PAGE and amino acid analysis. Maximum production of amino acids i.e. cystine, histidine and asparagine were observed in both the cow and soymilk after fermentation.

Production, purification, immobilization, and characterization of a thermostable ß-galactosidase from Aspergillus alliaceus.

A fungal strain isolated from rotten banana and identified as Aspergillus alliaceus was found capable of producing thermostable extracellular ß-galactosidase enzyme. Optimum cultural conditions for ß-galactosidase production by A. alliaceus were as follows: pH 4.5; temperature, 30 °C; inoculum age, 25 h; and fermentation time, 144 h. Optimum temperature, time, and pH for enzyme substrate reaction were found to be 45 °C, 20 min, and 7.2, respectively, for crude and partially purified enzyme. For immobilized enzyme-substrate reaction, these three variable, temperature, time, and pH were optimized at 50 °C, 40 min, and 7.2, respectively. Glucose was found to inhibit the enzyme activity. The K(m) values of partially purified and immobilized enzymes were 170 and 210 mM, respectively. Immobilized enzyme retained 43 % of the ß-galactosidase activity of partially purified enzyme. There was no significant loss of activity on storage of immobilized beads at 4 °C for 28 days. Immobilized enzyme retained 90 % of the initial activity after being used four times.

Application of principal component analysis (PCA) as a sensory assessment tool for fermented food products.

The objective of the work was to use the method of quantitative descriptive analysis (QDA) to describe the sensory attributes of the fermented food products prepared with the incorporation of lactic cultures. Panellists were selected and trained to evaluate various attributes specially color and appearance, body texture, flavor, overall acceptability and acidity of the fermented food products like cow milk curd and soymilk curd, idli, sauerkraut and probiotic ice cream. Principal component analysis (PCA) identified the six significant principal components that accounted for more than 90% of the variance in the sensory attribute data. Overall product quality was modelled as a function of principal components using multiple least squares regression (R (2) = 0.8). The result from PCA was statistically analyzed by analysis of variance (ANOVA). These findings demonstrate the utility of quantitative descriptive analysis for identifying and measuring the fermented food product attributes that are important for consumer acceptability.

Preparation of idli batter, its properties and nutritional improvement during fermentation.

Idli is a traditional fermented rice and black gram based food. Idli batter is prepared by soaking polished parboiled rice and decorticated black gram for 4 h at 30 ± 1 °C in water. The soaked mass was ground using a grinder with adequate amount of water. The blend ratios of 2:1, 3:1 and 4:1 (w/w) batter were allowed for fermentation for different periods with the addition of 2% (w/w) of salt. The rheology of the product was assessed using a Brookfield Viscometer having disc spindles. Shear stress values were in the range of 0.22 and 4 Pa and reached a maximum value at 7 h of fermentation. The density, pH, and percentage total acidity of batter during fermentation for different blend ratios ranged between 0.93 and 0.59 gm cm(-3), 4.21 and 5.9 and 0.44 and 0.91% respectively. During fermentation, maximum production of riboflavin and thiamine were found to be 0.76 mg/100 gm and 0.73 mg/100 gm in 3:1 blend ratio of idli batter; and the folic acid content was found to be at a maximum of 0.75 mg/100 gm of idli batter after 10 h of fermentation. Digestibility in terms of amino N2 content was analysed by formol titration.

Determination of kinetic parameters in the biosorption of Cr (VI) on immobilized Bacillus cereus M(1)(16) in a continuous packed bed column reactor.

Due to technological advancement, environment suffers from untreated toxic heavy metal bearing effluent coming from different industries. Chromium (VI) is one of those heavy metals having adverse impact on ecological balance, human, and plant health because of its carcinogenic properties. Biosorption is presented as an alternative to traditional technologies which are costly and inefficient for treatment of industrial wastes containing low amount of heavy metals. In this study, bioremediation of Cr (VI) ions by immobilized Bacillus cereus M(1) (16) was investigated in a laboratory scale packed bed up-flow column reactor. The effect of important parameters, such as the inlet flow rate, influent concentration, and effective bed height, has been studied. External mass transfer, surface adsorption, and intrabead mass transfer were also studied to conclude the rate limiting step for removal of Cr (VI) and to determine the process parameters which are important for biosorption optimization. The external mass transfer coefficient was calculated at different flow rates (6.51 x 10(-2) to 7.58 x 10(-2) cm/min). Using the model, the surface adsorption rate constant (k(ad)) and the intrabead mass transfer coefficient (k (i)) were predicted as 0.0267 x 10(-3) and 0.7465 x 10(-3) l/g/min, respectively. Both are much lower than the external mass transfer coefficient (k(e)). The surface adsorption phenomenon is acting as the rate-limiting step due to its high resistance for removal of Cr (VI).

Development of cholesterol biosensor based on immobilized cholesterol esterase and cholesterol oxidase on oxygen electrode for the determination of total cholesterol in food samples.

The development of a cholesterol biosensor by co-immobilization of cholesterol esterase (ChEt) and cholesterol oxidase (ChOX) on oxygen electrode is described. The electrode consists of gold cathode and Ag/AgCl anode. The enzymes were immobilized by cross-linking with glutaraldehyde and Bovine Serum Albumin (BSA). The immobilized enzymatic membrane was attached to the tip of the electrode by a push cap system. The optimum pH and temperature of the sensor was determined, these are 6 and 25 degrees C respectively. The developed sensor was calibrated from 1-75 mg/dl of cholesterol palmiate and found linear in the range of 2-50 mg/dL. The calibration curve was drawn with V(i) (ppm/min)(initial velocity) vs different concentrations of cholesterol palmiate (mg/dL). The application of the sensor to determine the total cholesterol in different real food samples such as egg, meat was investigated. The immobilized enzymatic layer can be reused over 30 times and the stability of the enzymatic layer was studied up to 9 weeks.

Development of biosensor based on immobilized L-glutamate oxidase for determination of monosodium glutamate in food.

A monosodium glutamate (MSG) biosensor with immobilized L-glutamate oxidase (L-GLOD) has been developed and studied for analysis of MSG in sauces, soup etc. The immobilized enzymatic membrane was attached with oxygen electrode with a push cap system. The detection limit of the sensor was 1 mg/dl and the standard curve was found to be linear upto 20 mg/dl. Response time of the sensor was 2 min. Cross-linking with glutaraldehyde in presence of Bovine Serum Albumin (BSA) as a spacer molecule has been used for immobilization. Optimization of the sensor was done with an increase in L-GLOD concentration (6.3-31.5 IU) and also with increase in loading volume of enzyme solution (5-20 microl). Optimization of pH and temperature was also studied. The permeability of O2 through different membrane was studied with and without immobilized L-GLOD. The enzymatic membrane was used for over 20 measurements and stability of the membrane was observed.

A biosensor based on co-immobilized L-glutamate oxidase and L-glutamate dehydrogenase for analysis of monosodium glutamate in food.

A monosodium glutamate (MSG) biosensor made by co-immobilized L-glutamate oxidase (L-GLOD) and L-glutamate dehydrogenase (L-GLDH) as the bio-component based on substrate recycling for highly sensitive MSG or L-glutamate determination, has been developed. Regeneration of MSG by substrate recycling provided an amplification of the sensor response. Higher signal amplification was found in the presence of ammonium ion. The sensor was standardized to determine MSG in the range of 0.02-3.0 mg/L. Linearity was obtained from 0.02 to 1.2 mg/L in presence of ammonium ion (10 mM) and NADPH (reduced nicotinamide adenine dinucleotide phosphate) (2 mM), but in absence of L-GLDH, the detection limit of MSG is confined to 0.1 mg/L. The apparent Km for MSG with L-GLOD-L-GLDH coupled reaction was 0.4451 mM but 1.9222 mM when only L-GLOD was immobilized. Cross linking with glutaraldehyde in the presence of bovine serum albumin (BSA) as a spacer molecule has been used for the method of immobilization. The response time of the sensor was 2 min. The optimum pH and temperature of the biosensor has been determined as 7+/-2 and 25+/-2 degrees C, respectively. The enzyme immobilized on the membrane was used for over 50 measurements. The standard error of the sample measurement was 4-5%. The activity of the enzyme-immobilized membrane was tested over a period of 60 days.

Development of an amperometric hypoxanthine biosensor for determination of hypoxanthine in fish and meat tissue.

A hypoxanthine (Hx) biosensor based on immobilized xanthine oxidase (XO) as the bio-component was developed and studied for the rapid analysis of fish (sweet water and marine) and goat meat samples. The biosensor was standardized for the determination of Hx in the range of 0.05 to 2 mM. Crosslinking with glutaraldehyde in presence of BSA as a spacer molecule was used for the method of immobilization. One layer of gelatin (10%) was applied over the immobilized enzyme layer to reduce the leaching out of enzyme from the membrane (cellulose acetate) matrix. The optimum pH of the immobilized system was determined to be 8.5 at 25 degrees C instead 7.0-7.2 for free enzyme system. Km and Vmax values were determined for the immobilized system. The developed sensor was applied to determine the amount of Hx present in fish and meat over a period of time. The stability of the enzyme immobilized membrane was also tested over a period of 30 days.

Production of a new antibacterial antibiotic from a mutated strain of Streptomyces kanamyceticus M27 in submerged fermentation.

A new antibacterial antibiotic was produced (136.5 microg/ml) using a 5 L EYELA Fermenter using 2 L fermentation medium at temperature: 27 degrees C, pH: 7.2, agitator speed: 200 rpm, aeration rate 1 vvm having KLa 251.74 hr(-1) at 96 hrs. The optimised conditions for antibiotic using washed cells of the selected strain are pH: 7.2, temperature: 27 degrees C, age of the biomass: 72 hr, amount of washed cell: 4 g in 50 ml normal saline, incubation time 72 hr. The antibacterial activity of the fermented broth was also examined against some bacterial species and it was found that it is active against gram positive as well as gram negative bacteria.

Production of lipase in a fermentor using a mutant strain of Corynebacterium species: its partial purification and immobilization.

Extracellular Corynebacterium lipase was produced using a 2.5 L Chemap fermentor using 1300 ml fermentation medium at temperature 33 degrees C, agitator speed 50 rpm, aeration rate 1 VVM having KLa 16.21 hr(-1). Crude lipase was purified by salting out method followed by dialysis and immobilized using calcium alginate gel matrix followed by glutaraldehyde cross linking Purification process increased specific activity of enzyme from 2.76 to 114.7 IU/mg. Activity of immobilized enzyme was 107.31 IU/mg. Optimum temperature for purified and immobilized enzyme activity were 65 degrees and 50 degrees C respectively. Optimum pH was 8.0 in both the cases, Km and Vmax value for purified lipase were 111.1 micromol/min and 14.7% respectively. Ca2+ (5 mM) was found to be stimulator for enzyme activity. Immobilized lipase retained 68.18% of the original activity when stored for 40 days.