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PubMedDate not supplied

Purification and characterization of aclacinomycin A and its metabolites from human urine.

Eleven metabolites of the anthracycline antineoplastic antibiotic aclacinomycin A (Acm) were isolated and purified from the urine of four patients treated with 100-120 mg/m2 of Acm. In an initial step, pooled urine was extracted and Acm and its metabolites were concentrated in n-butanol with a horizontal flow-through coil planet centrifuge. Individual metabolites were subsequently isolated and purified by thin-layer chromatography (TLC). Purified urinary species were characterized and compared to standards of Acm and its known metabolites by TLC, high performance liquid chromatography (HPLC), acid and enzymatic hydrolysis, and mass spectrometry. Nine of the urinary species were identified as either Acm or metabolites previously described in fermentation broths of Streptomyces galileus, in vitro reactions, animal plasma, or human plasma. Another urinary species corresponded to the major and previously unidentified Acm metabolite observed in human plasma. Sufficient quantitites of this material were isolated to allow its characterization and identification as bisanhydroaklavinic acid. The most polar urinary species isolated was identified as a beta-glucuronide conjugate of bisanhydroaklavinic acid. As such, these studies demonstrate the ability of humans to produce all of the previously proposed metabolites of Acm and establish the identity of the major Acm metabolite observed in human plasma.

Aclarubicin↗

PubMed1984

[Rapid method of determining beta-lactamase inhibitor activity].

A simple and rapid method for determination of the activity of beta-lactamase is described. It is based on spectrophotometric recording of the enzymatic splitting of benzylpenicillin. The method is recommended for rapid control of production of beta-lactamase inhibitors.

Bacillus↗

PubMed1980

Studies on terreic acid.

It was found that Aspergillus sp. No. Y-8980 which was isolated from a soil sample collected at Yoron Island in Kagoshima Prefecture belonged to Aspergillus terreus group by morphological observation. The active substance produced by the strain was obtained with a high yield in sucrose-yeast extract medium and extracted by chloroform, ethyl acetate and n-butanol at pH 2.4 approximately 2.6 from the culture broth. The substance was crystallized from chloroform and ethyl acetate after charcoal treatment of the crude crystal. From various physico-chemical properties, it was found that the substance was identical to terreic acid. Terreic acid showed MICs of 25 approximately 100 mcg/ml, 12.5 mcg/ml and 50 mcg/ml against Gram-positive and Gram-negative bacteria, Xanthomonas oryzae and Xanthomonas citri, respectively, but it did not control Pseudomonas, fungi and yeast. The LD50 was 75 mg/kg i.p. and i.v. in mice. With regards to the anti-tumor effect, the morphological degeneration on HeLa cells (human carcinoma cells) was observed in the concentrations of more than 6.25 mcg/ml of terreic acid. An increase of body weight of mice caused by Ehrlich ascites carcinoma cells was not definitely observed by the daily administration of 150 mcg of terreic acid per mouse for 8 consecutive days. Above showed the enough survival effect in dd mice implanted with Ehrlich ascites carcinoma cells, and the effect also was demonstrated by anatomies of mice.

Animals↗

PubMedDate not supplied

5-hydroxy-3-vinyl-2(5H)-furanone--a new inhibitor of human synovial phospholipase A2 and platelet aggregation from fermentations of a Calyptella species (basidiomycetes).

5-Hydroxy-3-vinyl-2(5H)-furanone, a potent and selective inhibitor of human synovial phospholipase A2 was isolated from fermentations of a Calyptella species. Its structure as identified by spectroscopic methods is identical to PA 147, an antibiotic previously isolated from a streptomycete. 5-hydroxy-3-vinyl-2(5H)-furanone inhibits the aggregation of human and bovine platelets stimulated by different inducers and exhibits weak antimicrobial activities.

4-Butyrolactone↗

PubMed2002

In vitro efficacies of phosphorolytic enzymes synthesized in mycelial cells of Aspergillus niger AbZ4 grown by a liquid surface fermentation.

Activities of phytase, a pH 6.0 optimum nonspecific phosphomonoesterase and phosphodiesterase assayed toward bis(p-nitrophenyl)phosphate (phosphodiesterase I) and against p-nitrophenylphosphorylcholine (phosphodiesterase II), were partially purified from mycelial extracts of Aspergillus niger AbZ4 cultivated on a molasses medium by a liquid surface fermentation method. After elimination of phosphate from the medium, 7.3- and 3.5-fold enhancements in specific activities of phytase and phosphodiesterase II were observed. Efficacies of mycelial protein fractions in dephosphorylating a wheat-based broiler feed were determined in vitro according to a procedure that simulated digestion in the intestinal tract of poultry. The addition of 0.052 mg of protein from fractions, each of which was high in either pH 6.0 optimum phosphomonoesterase, phosphodiesterase I, phosphodiesterase II, or phytase per gram of a feed sample resulted in the enhancement of phosphorus release by 10, 11, 27, and 88%, respectively. In the presence of an excess of commercial phytase, the addition of the mycelial fraction high in phytase increased the dephosphorylation rate by 56%. The fraction high in phosphodiesterase II enhanced feed dephosphorylation by 8% in the presence of an excess of commercial phytase and commercial acid phosphatase.

6-Phytase↗

PubMed1990

Evaluation of various nitrogen supplements in starter diets for growing Holstein steers and their effects on ruminal bacterial fermentation in continuous culture.

Concurrent in vivo and in vitro studies were conducted to evaluate urea (U), soybean meal (SBM), ground soybeans (RAW), extruded soybeans (ES) or extruded soybeans plus urea (ES + U) as primary supplemental N sources in starter diets for Holstein steers. Three groups of 48 Holstein steers each were fed five different starter diets to 181 kg BW in three experimental periods over 2 yr. Average daily gains were similar (P greater than .05) for steers fed ES + U (1.12 kg), ES (1.08 kg) and SBM (1.09 kg) but lower (P less than .05) for those fed U (1.00 kg) or RAW (.97 kg) diets. Feed/gain was similar (P greater than .05) for ES-fed steers vs those fed other diets except U. From 181 to 477 kg, all steers were fed the same diet. Steers fed the RAW starter diet had the lowest (P less than .05) ADG for the entire period. The starter diets were used as substrates for ruminal microbial metabolism in eight dual-flow continuous culture fermenters. True OM digestion was higher and NDF and ADF digestion was lower (P less than .05) for the ES + U diet than for the ES diet. Dietary protein degradation was lowest (P less than .05) for the ES diet (64.4%). Total bacterial N flow was higher (P less than .05) with the ES + U, SBM and U diets than with the ES diet. Lysine flow was higher (P less than .05) for the ES + U diet than for all other diets except ES. Results of these experiments indicate that ES as a protected ruminal escape N source with or without added urea did not improve steer performance above that obtained from SBM in starter diets.

Animal Feed↗

PubMed1997

A skin equivalent model for cosmetological trials: an in vitro efficacy study of a new biopeptide.

The European Community directive, imposing that effects claimed for cosmetic actives must be validated using non-animal procedures, has stimulated the use of in vitro models for pharmacotoxicological trials. In this paper, an efficacy study of a new biopeptide, a hydrolysate obtained by fermentation of milk proteins, was performed using an in vitro skin equivalent (SE). This SE is obtained by seeding normal human keratinocytes onto a dermal equivalent comprising a collagen-glycosaminoglycan(GAG)-chitosan porous matrix populated by normal human fibroblasts, which neosynthesize their own extracellular matrix (ECM). A gel containing 2% milk biopeptide was applied topically (10 microliters) every 2 days during 15 days. Subsequent investigations of the biopeptide effects were based on morphological criteria after histological analysis and on synthesis of ECM components. Collagen and GAG synthesis were measured by tritiated proline, glucosamine and Na2(35)SO4 incorporation. Qualitatively, the histological features of the biopeptide-treated SEs showed a thicker epidermis than the untreated control SEs, where only a few layers of stratum corneum were observed. The dermal porous matrix seems to be more filled by neosynthesized ECM than the control. Quantitatively, milk biopeptide treatment induced a significant activation of hyaluronic acid (+46%) and sulfated GAG (+53%) synthesis, whereas only non-significant increases of total protein and collagen synthesis were observed (Student's test, p < 0.001).

Animal Testing Alternatives↗

PubMed2001

Effect of protein source and soluble carbohydrate addition on rumen fermentation and lactation performance of Holstein cows.

Rumen in vitro and in vivo experiments were conducted to evaluate the effect of rumen undegradable protein and soluble carbohydrates on rumen ammonia N release and lactation performance of Holstein cows. In the in vitro experiment, freeze-dried annual ryegrass (Lolium multiflorum, LAM) pasture was supplemented 1:1 with ground corn-based grain supplements containing expeller or solvent soybean meal with sucrose or lactose supplements at 0, 2.5, or 5% of dry matter (DM). The ammonia release rate was slower with expeller compared with solvent soybean meal-supplemented diets. Sucrose supplementation at the 5% level lowered rumen ammonia concentrations, but lactose-fortification of grain supplements was without effect. In the in vivo study, 32 multiparous Holstein cows were blocked according to milk yield and randomly assigned to corn-based grain supplements containing 1) solvent soybean meal, 2) solvent soybean meal + 5% sucrose supplement, 3) expeller soybean meal, or 4) expeller soybean meal + 5% sucrose supplement. Grain supplements and fresh annual ryegrass were component fed at approximately a 1:1 grain to forage ratio (DM basis). Forage DM intake was higher for cows receiving solvent soybean meal supplemented grain supplements than those receiving expeller soybean meal (12.2 +/- 2.1 vs. 11.4 +/- 2.2 kg/d), but total DM intake was similar for all diets (22.8 +/- 2.9 kg/d). Fat-corrected milk yield was similar for all diets averaging 37.5, 38.2, 39.1, and 37.6 kg/d for diets 1 to 4, respectively. Rumen fermentation, milk urea nitrogen, and body condition were unaffected by supplements; however, cows fed grain supplement 1 utilized dietary energy more efficiently than cows offered the other dietary treatments. High dietary crude protein concentrations may have limited lactation response to rumen undegradable protein and sugar.

Ammonia↗

PubMed2002

Phenol toxicity and conjugation in human colonic epithelial cells.

BACKGROUND: Colonic epithelial cells are exposed to a range of potentially harmful luminal factors. including phenols, but it is unresolved whether these compounds impair the integrity of the epithelium. The aim of this study was to describe the effect of phenol exposure on human colonic epithelial cells in vitro and the conjugation pathways involved in detoxification. METHODS: Primary human colonic epithelial cell cultures or HT-29 cell cultures were exposed to paracetamol, dinitrophenol or phenol (0.1-5 mM) for 24 h. Cell viability was measured using the methyltetrazoleum test. Phenol conjugation products released from cell cultures were identified by high-pressure liquid chromatography. Phenol glucuronidase (PGD) and sulphotransferase (PST) enzyme activities were measured in isolated cell homogenates. RESULTS: Paracetamol, dinitrophenol and phenol (>1.25 mM) significantly impaired the viability of primary colonic epithelial cell cultures. No differences between cell cultures from ulcerative colitis and control patients were observed. Paracetamol (5 mM) also induced significant cell damage in HT-29 cells. Glucuronidation was the preferred conjugation pathway in both cell models, despite the presence of PGD and PST activity. CONCLUSION: Phenols have a direct toxic effect on human colonic epithelial cells in vitro, which supports the view that dietary fermentation metabolites may be involved in the modulation of chronic bowel inflammation.

Adult↗

PubMed2002

Chemopreventive and adjuvant therapeutic potential of pomegranate (Punica granatum) for human breast cancer.

Fresh organically grown pomegranates (Punica granatum L.) of the Wonderful cultivar were processed into three components: fermented juice, aqueous pericarp extract and cold-pressed or supercritical CO2-extracted seed oil. Exposure to additional solvents yielded polyphenol-rich fractions ('polyphenols') from each of the three components. Their actions, and of the crude whole oil and crude fermented and unfermented juice concentrate, were assessed in vitro for possible chemopreventive or adjuvant therapeutic potential in human breast cancer. The ability to effect a blockade of endogenous active estrogen biosynthesis was shown by polyphenols from fermented juice, pericarp, and oil, which inhibited aromatase activity by 60-80%. Fermented juice and pericarp polyphenols, and whole seed oil, inhibited 17-beta-hydroxysteroid dehydrogenase Type 1 from 34 to 79%, at concentrations ranging from 100 to 1,000 microg/ml according to seed oil >> fermented juice polyphenols > pericarp polyphenols. In a yeast estrogen screen (YES) lyophilized fresh pomegranate juice effected a 55% inhibition of the estrogenic activity of 17-beta-estradiol; whereas the lyophilized juice by itself displayed only minimal estrogenic action. Inhibition of cell lines by fermented juice and pericarp polyphenols was according to estrogen-dependent (MCF-7) >> estrogen-independent (MB-MDA-231) > normal human breast epithelial cells (MCF-10A). In both MCF-7 and MB-MDA-231 cells, fermented pomegranate juice polyphenols consistently showed about twice the anti-proliferative effect as fresh pomegranate juice polyphenols. Pomegranate seed oil effected 90% inhibition of proliferation of MCF-7 at 100 microg/ml medium, 75% inhibition of invasion of MCF-7 across a Matrigel membrane at 10 microg/ml, and 54% apoptosis in MDA-MB-435 estrogen receptor negative metastatic human breast cancer cells at 50 microg/ml. In a murine mammary gland organ culture, fermented juice polyphenols effected 47% inhibition of cancerous lesion formation induced by the carcinogen 7,12-dimethylbenz[a]anthracene (DMBA). The findings suggest that clinical trials to further assess chemopreventive and adjuvant therapeutic applications of pomegranate in human breast cancer may be warranted.

17-Hydroxysteroid Dehydrogenases↗

PubMed2003

Invasion of Salmonella enteritidis in avian intestinal epithelial cells in vitro is influenced by short-chain fatty acids.

Fermentation reactions in the caeca of chickens, the predominant place for Salmonella colonization, result in high concentrations of short-chain fatty acids (SCFA). Thus Salmonella bacteria are in close contact with SCFA during their life cycle. A study was carried out to analyse the effects of SCFA on invasion of Salmonella enteritidis in an avian intestinal epithelial cell line. Preincubation of S. enteritidis for 4 h in growth media supplemented with various concentrations of propionate or butyrate resulted in decreased invasion compared to bacteria, preincubated in nonsupplemented media, and to bacteria, preincubated in media supplemented with formate or acetate. Incubation of the S. enteritidis bacteria in media supplemented with mixtures of SCFA mimicking the in vivo caecal concentrations resulted in increased invasion compared with butyrate-exposed bacteria, but equal invasion compared with nonexposed bacteria. Increasing the butyrate concentration in these mixtures did not modify invasion compared with the original mixtures.

Animals↗

PubMed1994

Effects of ruminal protozoa on cellulose degradation and the growth of an anaerobic ruminal fungus, Piromyces sp. strain OTS1, in vitro.

An anaerobic rumen fungus, Piromyces sp. strain OTS1, was incubated in the presence or absence of a mixed, A-type, protozoal population obtained from a goat, in a medium containing filter paper cellulose as energy source and antibiotics to suppress bacterial growth. Fermentation end products, cellulose degradation, and chitin as an indicator of fungal biomass were examined. In the presence of protozoa, total volatile fatty acids, notably propionate and butyrate, increased, and lactate decreased. In fungus-protozoan coincubations, formate was not detected at the end of the experiment and the amount of reducing sugars remained low throughout the incubation period. The fungal growth in the coincubations was negatively affected. While protozoal predation on zoospores was one mechanism of inhibition, mature fungal cells were also affected. Total cellulose degradation was greater in fungal monocultures, but the amount of cellulose degraded per unit of fungal biomass was 25% larger in the coincubations. The negative effects that the protozoal predatory activity had on the fungal growth and subsequently on the amount of cellulose degraded by Piromyces sp. strain OTS1 were partially attenuated by the protozoal fibrolytic activity or by an enhanced fungal activity due to a more favorable environment.

Anaerobiosis↗

PubMed1985

Short-chain fatty acids in intestinal content of germfree mice monocontaminated with Escherichia coli or Clostridium difficile.

The short-chain fatty acids (SCFAs) have been analysed in coecal and small-intestinal content of conventional (CONV) and germfree (GF) mice, in germfree mice monocontaminated with Escherichia coli (MEC) or Clostridium difficile (MCD), and in germfree mice conventionalized by the visitor technique (EXG). The total concentrations of SCFAs in coecal content, measured by gas chromatography, were (mean (SD), mmol/kg): CONV, 125.2 (32.9); GF, 1.02 (0.39); MEC, 6.88 (0.76); MCD, 4.50 (0.12); and EXG, 115.6 (17.4). The concentrations of SCFAs were 3- to 25-fold higher in the coecum than in the small intestine in CONV, MEC, and EXG mice (p less than 0.05). The fermentation patterns (that is, the relative composition of the acids) of E. coli and Cl. difficile were distinctively different under defined in vitro conditions and similar to those found in intestinal content of monocontaminated animals. Combined gas chromatography and mass spectrometry showed that Cl. difficile produced an unusual metabolite, 2-methylbutyric acid, in vitro and in vivo. The findings indicate that the fermentation patterns are closely related to the bacteria. Variations in the bacterial flora may be more important in determining the concentrations and patterns of SCFAs in intestinal content than variations in the intake of substrate for SCFAs formation as dietary fibre.

Animals↗

PubMedDate not supplied

[Effect of trace metals on cell morphology, enzyme activation, and production of citric acid in a strain of Aspergillus wentii].

Data concerning the effect of very low concentrations of metals on citric acid production by microorganisms, as well as on the activity of enzymes presumptively involved in the process, are confuse. The bulk of information was obtained mainly studying selected strains of Aspergillus niger. Information concerning other citric acid producer filamentous fungi, such as A. wentii, is scanty. In the present article we report the effect of different cations on the growth pattern of A. wentii P1 as well as on the related citric acid production and the activity of several enzymes. It was found that without any addition to the culture medium the fungus developed a pelleted form of growth, pellets being about 1.5 mm in diameter. The citric acid yield was about 90%. The addition of Cu2+ impaired the sugar uptake, as well as the production of citric acid and biomass. The uptake of sugar increased in the presence of Zn2+, and there was a marked increase of the biomass production, which could account for the low citric acid production. The addition of Fe2+ impaired the citric acid production and, as sulfate, the sugar uptake. The presence of Fe3+ markedly impaired the citric acid production and increased the sugar uptake. There is no agreement about the enzymes involved in the accumulation of citric acid by microorganisms. In spite of this, aconitase (Ac), isocitrate lyase (IL), isocitrate dehydrogenase NAD(+)-dependent (ICDH- NAD+) and isocitrate dehydrogenase NADP(+)-dependent (ICDH-NADP+) are often postulated as key enzymes. In our case, these enzymes were active during the standard fermentation, although with variations, particularly concerning Ac and IL. The behavior of enzymes might be different when tested in vivo or in vitro, mainly from the quantitative point of view. Nevertheless, the activity determined in vitro might give some indication concerning the effect on fermentation of substances present in the medium. It was found that all the enzymes tested increased their activity in the presence of Fe2+ and Fe3+. The addition of Cu2+ improved the activity of Ac, IL and ICDH-NADP+, while that of ICDH-NAD+ was impaired. The presence of Zn2+ decreased the activity of the enzymes excepting that of ICDH-NADP+ which increased. It must be pointed out that according to these results there are differences concerning the behavior of ICDH-NAD+ and ICDH-NADP+ in the presence of Cu2+ or Zn2+.

Aconitate Hydratase↗

PubMedDate not supplied

In vitro and in vivo assessment of intraintestinal bacteriotherapy in chronic kidney disease.

Chronic kidney disease may progress to end-stage renal disease, which requires dialysis or kidney transplantation. No generally applicable therapies to slow progression of renal disease are available. Bacteriotherapy affords a promising approach to mitigate uremic intoxication by ingestion of live microbes able to catabolize uremic solutes in the gut. The present study evaluates the nonpathogenic soil-borne alkalophilic urease-positive bacterium Sporosarcina pasteurii (Sp) as a potential urea-targeted component for such "enteric dialysis" formulation. Data presented herein suggest that Sp survives through exposure to gastric juice retaining the ability to hydrolyze urea. In vitro, 10 cfu (colony forming units) of Sp removed from 21 +/- 4.7 mg to 228 +/- 6.7 mg urea per hour, depending on pH, urea concentration, and nutrient availability. Beneficial effects of Sp on fermentation parameters in the intestine were demonstrated in vitro in the Simulator of the Human Intestinal Microbial Ecosystem (SHIME) inoculated with fecal microbiota. Enumeration of marker organisms suggested that presence of Sp does not disturb microbial community of the SHIME. Additionally, a pilot study in 5/6th nephrectomized rats fed 10 cfu of live Sp daily throughout the study demonstrated that the tested regimen reduced blood urea-nitrogen levels and significantly prolonged the lifespan of uremic animals.

Animals↗

PubMed1997

Administration of transgalacto-oligosaccharides increases fecal bifidobacteria and modifies colonic fermentation metabolism in healthy humans.

Transgalacto-oligosaccharides are a mixture of oligosaccharides consisting of glucose and galactose; they are not digested in the human small intestine. In vitro, they specifically stimulate the growth of bifidobacteria. The aim of the present work was to assess tolerance of transgalacto-oligosaccharides and the effects of their prolonged administration on bifidobacteria and fermentative activity of colonic flora. Eight healthy subjects were given 10 g of transgalacto-oligosaccharides per day for 21 d in two daily doses. A breath test and stool sample collection were carried out on d 1, 7, 14 and 21 of transgalacto-oligosaccharides ingestion. The stools of three subjects were collected and mixed before the study, and then inoculated in vitro into a fermentor to which 10 g transgalacto-oligosaccharides was added daily for 14 d. In the eight volunteers, administration of transgalacto-oligosaccharides led to a significant decrease in breath hydrogen excretion (P < 0.01) and a significant increase in fecal concentrations of bifidobacteria from (means +/- SEM) 8.6 +/- 0.6 to 9.7 +/- 0.5, 9.7 +/- 0.6 and 9.5 +/- 0.6 log colony-forming units (CFU)/g on d 1, 7, 14 and 21, respectively (P < 0.05). Fecal concentrations of enterobacteria, as well as stool weight, fecal water and pH did not change during the study. In vitro, transgalacto-oligosaccharides fermentation became more efficient and faster with time. In addition, metabolic alterations such as a rise in acetate proportion and lactate formation after 7 d of fermentation were observed, indicating the transformation of the inoculated fecal flora into an acid-resistant lactic flora. Prolonged administration of transgalacto-oligosaccharides, at a dose which does not induce digestive symptoms, increases the number of bifidobacteria and alters the fermentative activity of colonic flora in humans.

Adult↗

PubMed1996

Release of cytokines by a fermented lectin-1 (ML-1) free mistletoe extract reflects differences in the reactivity of PBMC in healthy and allergic individuals and tumour patients.

OBJECTIVE: Mistletoe extracts are used in adjuvant cancer treatment, but little is known concerning their mode of action. There is, however, evidence that antigens in these extracts may stimulate cells of the immune system, thereby modifying the altered immunological reactivity in tumour patients. METHODS: In order to find out whether the postulated immunomodulatory properties of mistletoe extracts are mediated by cytokines, a spectrum of different cytokines was analysed in the supernatants of peripheral blood mononuclear cells (PBMC) from healthy (n = 23) and allergic (n = 16) individuals after stimulation with the fermented mistletoe lectin-1 (ML-1) free mistletoe extract Iscador Pini (IP) in vitro, and their cytokine patterns were compared to those from tumour patients with either breast cancer (n = 20) or colorectal tumours (n = 22). RESULTS: PBMC from healthy and allergic individuals produced high levels of TNF-alpha and IL-6 and to a lesser extent Th1- and Th2-related cytokines. This finding was in contrast to data obtained in tumour patients. Thus, the concentration of TNF-alpha was significantly lower in the cell cultures from breast cancer patients than in controls, and patients with colorectal tumours released IFN-gamma/IL-2 (5%) in the supernatants significantly less frequently than PBMC from healthy controls (26%). Similar results were obtained when the Th1- and monocyte/macrophage-related cytokines were analysed in the unstimulated cell cultures. CONCLUSIONS: These in vitro studies provide evidence that there is a reduced immunological reactivity to the fermented ML-1 free mistletoe extract in tumour patients and may give some clues as to how mistletoe-derived antigens could act on immune cells involved in the tumour defence.

Adult↗

PubMed1999

Enzymatic treatment and use of starters for the nutrient enhancement in fermented flour of red and white varieties of finger millet (Eleusine coracana).

Two varieties of finger millet (Eleusine coracana)-a tannin-containing red variety, CO13, and nontannin white variety, CO9-processed by treatment with enzymes (cellulase and hemicellulase) and fermentation with starters (from previously fermented finger millet batter), achieved the desirable goals of reduced fermentation time (12 h), increased acidity (2.2 to 2.4%), enhanced in vitro protein digestibility (IVPD) (14 to 26%), and mineral availability compared to 48 h uncontrolled natural fermentation (Usha Antony and Chandra, 1998). Fermentation with starters alone increased titratable acidity (1.02 to 1.88%), IVPD (5. 5 to 22%) and mineral availability, and decreased phytate (23 to 26%) and tannin (10.8 to 40.5%) in the millets. Enzymatic treatment (3 h, 50 degrees C) did not significantly alter the pH, phytate, tannins, IVPD, or HCl-mineral extractability but enhanced fermentative changes. Overall, the changes were marked when the 48 h starter was used and the improvements in nutrient availability was greater in the CO13 variety.

Cellulase↗