Ethanol-Mediated Variations in Cellular Fatty Acid Composition and Protein Profiles of Two Genotypically Different Strains of Escherichia coli O157:H7

Two strains of Escherichia coli O157:H7 were grown in tryptic soy broth (TSB, pH 7.1) supplemented with 0, 2.5, 5.0, 7.5, and 10% ethanol at 30°C for up to 54 h. Growth rates in TSB supplemented with 0, 2.5, and 5.0% ethanol decreased with an increase in ethanol concentration. Growth was not observed in TSB supplemented with 7.5 or 10% ethanol. The pH of TSB containing 5.0% ethanol decreased to 5.8 within 12 h and then increased to 7.0 at 54 h. The ethanol content in TSB supplemented with 2.5 or 5.0% ethanol did not change substantially during the first 36 h of incubation but decreased slightly thereafter, indicating utilization or degradation of ethanol by both strains. Glucose was depleted in TSB supplemented with 0, 2.5, or 5.0% ethanol within 12 h. Cells grown under ethanol stress contained a higher amount of fatty acids. With the exceptions of cis-oleic acid and nonadecanoic acid, larger amounts of fatty acid were present in stationary-phase cells of the two strains grown in TSB supplemented with 5.0% ethanol for 30 h than in cells grown in TSB without ethanol for 22 h. The trans-oleic acid content was 10-fold higher in the cells grown in TSB with 5.0% ethanol than those grown in TSB without ethanol. In contrast, cis-oleic acid was not detected in ethanol-stressed cells but was present at concentrations of 0.32 and 0.36 mg/g of cells of the two strains grown in TSB without ethanol. Protein content was higher in ethanol-stressed cells than in nonstressed cells. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein profiles varied qualitatively as affected by the strain and the presence of ethanol in TSB. An ethanol-mediated protein (28 kDa) was observed in the ethanol-stressed cells but not in control cells. It is concluded that the two test strains of E. coli O157:H7 underwent phenotypic modifications in cellular fatty acid composition and protein profiles in response to ethanol stress. The potential for cross protection against subsequent stresses applied in food preservation technologies as a result of these changes is under investigation.     

The Relation between Accumulation of Abscisic Acid and Proline in Detached Rice Leaves

The relation between abscisic acid (ABA) and proline accumulation was investigated in detached rice (Oryza sativa L.) leaves. In darkness, proline content increased about 2-, 2,5- and 6-fold after 24, 48 and 72 h. ABA content reached maximum after 48 h. In the light, proline content remained almost unchanged until 48 h and subsequently increased slightly. ABA content in the light was lower than in darkness, but the maximum was also after 48 h. During 12-h exposure to decreased air humidity, proline content gradually increased, but ABA content increased about 25-fold after 4 h and declined thereafter. Exogenous application of ABA resulted in an increase in proline content in detached rice leaves under both light and darkness.     

Effect of vitamin C administration on blood cholesterol level in man

Vitamin C (1.0 g/day) was administered orally to 20 healthy males for 1 month under controlled conditions. The blood ascorbic acid level rose from 0.76 +/- 0.21 mg% to 1.24 +/- 0.19 mg% in young subjects (20-30 years), and from 0.74 +/- 0.29 mg% to 1.22 +/- 0.22 mg% in middle-aged ones (31-50 years). Simultaneously, the serum cholesterol levels decreased from 204 +/- 16 mg% to 177 +/- 21 mg% in the young and from 256 +/- 11 mg% to 225 +/- 36 mg% in the middle-aged, a statistically significant fall of 10-15%, on the average (P less than 0.01). The effect in normo-cholesteraemic subjects, in particular, supports the cholesterol-lowering action of vitamin C.     

Ethanol-mediated variations in cellular fatty acid composition and protein profiles of two genotypically different strains of Escherichia coli O157:H7

Two strains of Escherichia coli O157:H7 were grown in tryptic soy broth (TSB, pH 7.1) supplemented with 0, 2.5, 5.0, 7.5, and 10% ethanol at 30 degrees C for up to 54 h. Growth rates in TSB supplemented with 0, 2.5, and 5.0% ethanol decreased with an increase in ethanol concentration. Growth was not observed in TSB supplemented with 7.5 or 10% ethanol. The pH of TSB containing 5.0% ethanol decreased to 5.8 within 12 h and then increased to 7.0 at 54 h. The ethanol content in TSB supplemented with 2.5 or 5.0% ethanol did not change substantially during the first 36 h of incubation but decreased slightly thereafter, indicating utilization or degradation of ethanol by both strains. Glucose was depleted in TSB supplemented with 0, 2.5, or 5.0% ethanol within 12 h. Cells grown under ethanol stress contained a higher amount of fatty acids. With the exceptions of cis-oleic acid and nonadecanoic acid, larger amounts of fatty acid were present in stationary-phase cells of the two strains grown in TSB supplemented with 5.0% ethanol for 30 h than in cells grown in TSB without ethanol for 22 h. The trans-oleic acid content was 10-fold higher in the cells grown in TSB with 5.0% ethanol than those grown in TSB without ethanol. In contrast, cis-oleic acid was not detected in ethanol-stressed cells but was present at concentrations of 0.32 and 0.36 mg/g of cells of the two strains grown in TSB without ethanol. Protein content was higher in ethanol-stressed cells than in nonstressed cells. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein profiles varied qualitatively as affected by the strain and the presence of ethanol in TSB. An ethanol-mediated protein (28 kDa) was observed in the ethanol-stressed cells but not in control cells. It is concluded that the two test strains of E. coli O157:H7 underwent phenotypic modifications in cellular fatty acid composition and protein profiles in response to ethanol stress. The potential for cross protection against subsequent stresses applied in food preservation technologies as a result of these changes is under investigation.     

Temperature dependence-independence of antifreeze turnover in Eurosta solidaginis (Fitch)

Temperature effects on antifreeze metabolism were investigated in two populations (northern and southern) of the golden rod gallfly, Eurosta solidaginis. Sorbitol production was temperature dependent and was triggered by short-term exposure to < +10°C. The maximal rate sorbitol synthesis occurred at 0°C. For both populations, sorbitol was rapidly catabolized during warm acclimation at +20°C. During the first 12 h of warm acclimation, sorbitol levels decreased by 36% (19.7 ± 0.6) to 12.6 ± 1.2 μg/mg) and by 83% (to 3.3 ± 1.7 μg/mg) after 48 h in the northern population. The southern population decreased sorbitol levels 64% (11.8 ± 0.69 to 4.2 ± 0.62) after 48 h. The southern population resynthesized more sorbitol than did the northern population upon re-exposure to 0°C. Glycerol levels increased linearly during the experimental period independent of temperature.     

Self-limiting reflex luteinizing hormone release and sexual behavior during extended periods of unrestricted copulatory activity in estrous domestic cats

Estrous female cats (queens) were permitted 36-h periods of unrestricted mating activity; they then were injected with various doses of gonadotropin-releasing hormone (GnRH) at 36 h and allowed single copulations at 48 or 72 h of study. Serum luteinizing hormone (LH) levels were determined in samples collected prior to and 2 h after the initial copulation, before and 30 min after selected copulations during the next 10 h, and before and 30 min after copulations occurring at 20-24, 36, and 48-72 h, as well as 0, 15, and 30 min after the GnRH injections (0.3-3.0 micrograms/kg) at 36 h of study. Copulations occurred 14-20 times in 12 h and 20-36 times in 36 h. Copulation frequency (mean +/- SEM) decreased (p less than 0.05) from 5.5 +/- 0.6/2 h initially to 1.5 +/- 0.6/2 h during the subsequent 2-h period, and was 1.4 +/- 0.2/2 h at 12-36 h of study. Intromissions lasted 1-27 (8 +/- 0.3) s. Variation in durations of mounting by males (1.7 +/- 0.1 min; range, 0.3-10 min) or of the postcoital behavioral reactions displayed by the queens (2.5 +/- 0.1 min; range, 1-17 min) could not be related to animals or time of study. Peak serum LH levels (11-280 ng/ml; mean, 112 +/- 30 ng/ml) were observed at 2-4 h after the first mating. Mean LH steadily declined thereafter, reached basal values (less than or equal to 3 ng/ml) by 20-24 h, and remained low at 36 h.(ABSTRACT TRUNCATED AT 250 WORDS)     

Physiological changes in asparagus spear tips after harvest

To extend our understanding of the physiology of asparagus after harvest, changes in respiration rate, protein and amino acid complement, and ultrastructure of tip sections (0–30 mm) of asparagus spears ( Asparagus officinalis L. cv. Limbras 10) were investigated. Spears had been stored for up to 4 days in the dark at 20°C. Respiration rate (carbon dioxide efflux) declined rapidly after harvest before stabilizing at 12 h at ca 50% of the rate at harvest. Protein, amino acid, and ammonium content of tip sections of 180 mm spears (intact tip sections) during storage, and comparable sections; excised from spears at harvest and subsequently stored (excised tip sections), were compared. Total protein content of intact and excised tip sections increased ca 10% 6–12 h after harvest, and then declined to ca 85% of harvest levels at 48 h. Gel electrophoresis in the presence of sodium dodecyl sulphate revealed the net loss of specific proteins at 48 h. Free amino acid content of excised tip sections declined to ca 75% of harvest levels 12 h after harvest, and then increased to 150% of harvest levels by 48 h. Glutamine levels declined rapidly after harvest, and asparagine content increased ca 200% at 24 h. Similar trends in free amino acid content were found in sections of intact tips. Ammonia (ammonium ions) accumulated to ca 0.3% dry weight at 48 h in both intact and excised tip sections. Ultrastructural studies revealed that tonoplast breakdown commenced 48–96 h after harvest. Results are discussed in relation to the sequence of physiological events following harvest and the timing of mechanisms responsible for their initiation.     

Uptake, Distribution, and Metabolism of 1-Naphthaleneacetic Acid and Indole-3-Acetic Acid During Callus Initiation From Actinidia deliciosa Tissues

1-Naphthaleneacetic acid (NAA) and 6-benzyladenine (BA) were required for in vitro callus formation at the basal edge of kiwifruit (Actinidia deliciosa [A. Chev] Liang and Ferguson, cv. Hayward) petioles. The uptake, metabolism, and concentration of NAA and indole-3-acetic acid (IAA) content were examined in the explants during the callus initiation period. After 1, 6, 12, 24, 48, and 96 h of culture in the presence of [H³]NAA, petioles were divided into apical, middle, and basal portions and analyzed. Except for a high IAA level measured at 12 h, IAA content decreased in tissues during a culture period of 96 h. NAA uptake was higher in petiolar edges than in the middle portion, and NAA was rapidly conjugated with sugars and aspartic acid inside the tissues. The amide conjugation was triggered in apical and basal portions from 12 h and in the middle part from 48 h, with α-naphthylacetylaspartic acid being the major metabolite. Free-NAA concentration in cultured petioles achieved an equilibrium with the exogenously applied NAA (0.27 μm) from 12 h, and it remained constant thereafter. The relationships between the role attributed to NAA and BA in the initiation and the maintenance of disorganized growth of callus in kiwifruit cultures are discussed. Received December 21, 1998; accepted July 20, 1999     

Demineralization of crab shells by chemical and biological treatments

To achieve demineralization of crab shell waste by chemical and biological treatments, lactic acid and lactic acid bacterium were applied. In 5.0 and 10% lactic acid, pH rapidly decreased from 6.8 to 4.2 and from 4.5 to 2.4 at day 3, respectively, and thereafter the pH remained at an almost constant level. In a 10% lactic acid bacterium inoculum, pH lowered to 4.6 at day 5. Relative residual ash content rapidly decreased to 49.1 and 16.4% in 5 and 10% lactic acid treatments, respectively, for the initial 12 h. In 2.5, 5 and 10% lactic acid bacterium inoculums, relative residual ash content rapidly decreased to 55.2, 40.9 and 44.7%, respectively, on the first day. Residual dry masses were 76.4, 67.8 and 46.6% in 2.5, 5 and 10% lactic acid treatments, respectively, for the initial 12 h. After a one-time exchange of the lactic acid solution, in the 5.0% lactic acid treatment, residual dry mass rapidly decreased from 66.0 to 41.4%. In 2.5, 5 and 10% lactic acid bacterium inoculums, residual dry masses decreased to 67.6, 57.4 and 59.6% respectively, on the first day. Protein contents after demineralization ranged from 51.3–54.7% in the chemical treatments and decreased to 32.3% in the lactic acid fermentation process. A negative relationship was shown between pH and demineralization rate in lactic acid and lactic acid bacterium treatments. These results suggest that lactic acid fermentation can be an alternative for demineralization of crab shells, even though the rate and efficiency of the demineralization is lower than the chemical treatment.     

Effects of superovulatory doses of pregnant mare serum gonadotropin on oocyte quality and ovulatory and steroid hormone responses in rats

Immature rats (aged 28 days) were injected with 4, 20, or 40 IU pregnant mare serum gonadotropin (PMSG) and sacrificed every 6 or 12 hr. Control rats (4 IU) ovulated between 60 and 72 hr, whereas rats given superovulatory doses of PMSG (20 and 40 IU) ovulated between 24 and 72 hr. The oocyte count from the superovulated rats increased slightly between 24 and 36 hr and markedly between 48 and 72 hr. Degenerated oocytes were recovered 48 and 36 hr after administration of 20 and 40 IU PMSG, respectively. Thereafter, the proportion of degenerated oocytes was dose dependent and reached a maximum at 72 (30.9%, 20 IU) and 60 hr (61.0%, 40 IU). 17β-estradiol content of the superovulated ovaries increased significantly (P < 0.01) from 36 hr and was maximal at 60 (20 IU) or 54 hr (40 IU), when compared to the control regimen. Administration of 40 IU PMSG resulted in a biphasic increase of progesterone content with the peaks at 36 and 60 hr. Androgen content of the superovulated ovaries was lower than control levels during the first 36 hr but was significantly (P < 0.01) higher thereafter. The results suggest that these alterations in the steroid response (particularly androgens) from 36 hr onward following superovulation may be responsible for the coincidental occurrence of abnormal oocytes, possibly by disturbing the specific intrafollicular steroid environment essential for complete maturation. In addition, oocyte aging that is due to earlier activation by the exogenous luteinizing hormone activity may be a contributing factor.     

Expression of dehydrins under heat stress and their relationship with water relations of sugarcane leaves

The heat stress-induced dehydrin proteins (DHNs) expression and their relationship with the water relations of sugarcane (Saccharum officinarum L.) leaves were studied. Sugarcane seedlings were subjected to heat stress (day/night temperature of 40/35 °C) under relative humidity 60/65 % to avoid aerial desiccation and determinations made at 4, 12, 24, 36, 48, 60 and 72 h. The leaves showed a sharp decline in the water and osmotic potentials, and relative water content during first 12 h of heat stress, but a regain in their values in 24 h. The pressure potential (ψ_p) decreased initially but increased later and approached control leaves. The increase in ψ_p was tightly correlated to the accumulation of free proline, glycinebetaine and soluble sugars, indicating their possible involvement in the osmotic adjustment under heat stress. Immunological detection revealed the expression of three DHNs with an apparent molecular mass of 21, 23 and 27 kDa under heat stress (48 to 72 h) and their expression was independent of the changes in the water relations of leaves.     

Mild, solvent-free omega-hydroxy acid polycondensations catalyzed by candida antarctica lipase B

Immobilized Candida antarctica Lipase B (Novozyme-435) was studied for bulk polyesterifications of linear aliphatic hydroxyacids of variable chain length. The products formed were not fractionated by precipitation. The relative reactivity of the hydroxyacids was l6-hydroxyhexadecanoic acid approximately 12-hydroxydodecanoic acid approximately 10-hydroxydecanoic acid (DPavg congruent with 120, Mw/Mn 6-hydroxyhexanoic acid (DPavg congruent with 80, Mw/Mn < or = 1.5, 48 h, 90 degrees C). Remarkable improvements in molecular-weight buildup resulted from leaving water in the reaction. By 4 h, without application of vacuum, the DPavg for 12- and 16-carbon hydroxyacids was about 90. In contrast, with identical substrates and water removal, the DPavg at 4 h was about 23. Large differences in the molecular-weight build up of 12-hydroxydodecanoic acid were observed for catalyst concentrations (%-by-wt relative to monomer) of 0.1, 0.5, 1, and 10. Nevertheless, by 24 h, with 1% catalyst containing 0.1% lipase, poly(12-hydroxydodecanoic acid) with Mn 17 600 was formed. For 12-hydroxydodecanoic acid polymerization at 90 degrees C, the catalyst activity decreased by 7, 18, and 25% at reaction times of 4, 24, and 48 h, respectively. Furthermore, the retention of catalyst activity was invariable as a function of the substrates used.     

培养基中能量底物对猪胚胎发育的影响

旨在探讨丙酮酸和乳酸对猪(Susscrofa)胚胎早期发育的影响,将NCSU-23培养基中的5.56mmol/L葡萄糖替换为0.2mmol/L丙酮酸、5.7mmol/L乳酸,并将此培养基命名为mNCSU-23。根据实验设计,孤雌胚及核移植胚转移到mNCSU-23或NCSU-23中培养。激活第2天统计孤雌胚及核移植胚中的5～8细胞胚胎数。激活第6天统计孤雌胚及核移植胚囊胚形成率及囊胚细胞数。实验结果表明,mNCSU/NCSU处理组的5～8细胞胚胎数及囊胚数显著高于对照组(P0.05);单纯使用mNCSU培养猪胚胎时,囊胚率最低,发育结果最差(P0.05)。本研究证实,在体外培养前两天,用乳酸和丙酮酸代替培养基中的葡萄糖对胚胎发育有利。     

Effect of the engineered indole pathway on accumulation of phenolic compounds in Catharanthus roseus hairy roots

Catharanthus roseus has been well-known to contain indole alkaloids effective for treatment of diverse cancers. We examined the intracellular accumulation profiles of phenolic compounds in response to ectopic overexpression of tryptophan feedback-resistant anthranilate synthase holoenzyme (ASalphabeta) in C. roseus hairy roots. Among 13 phenolic compounds measured, 6 phenolic compounds were detected in late exponential phase ASalphabeta hairy roots. Uninduced and induced ASalphabeta hairy roots accumulated up to 1.2 and 4.5 mg/g DW over a 72-h period, respectively. Upon induction, in parallel with a rapid increase in tryptophan in the first 48 h, accumulation of phenolic compounds tended to increase to a maximum level (4.5 mg/g DW) at 48 h, after which phenolic levels decreased back to the uninduced level by 72 h. Naringin was a predominant form that comprised about 72% and 36% of the total content of phenolic compounds in the uninduced and induced lines, respectively. Upon induction, accumulation of catechin drastically increased with the highest level (3.6 mg/g) occurring at 48 h, whereas that of all others except for salicylic acid showed no statistical difference. Catechin is a final product of the flavonoid pathway, and thus metabolic flux into this pathway is transiently increased by overexpression of AS. Like catechin, salicylic acid is very sensitive to induction as it began to increase to 5-fold within 4 h of induction, but unlike catechin, no significant accumulation of salicylic acid was noted after 4 h of induction. The results suggest differential regulation of this particular biosynthesis branch within the phenolic pathway.     

Alteration in Fatty Acid Composition of Adult Rat Brain Capillaries and Choroid Plexus Induced by a Diet Deficient in n-3 Fatty Acids: Slow Recovery After Substitution with a Nondeficient Diet

Wistar rats were fed for three generations with a semisynthetic diet containing either 1.5% sunflower oil (940 mg% of C18:2n-6, 6 mg% of C18:3n-3) or 1.9% soya oil (940 mg% of C18:2n-6, 130 mg% of C18:3n-3). At 60 days of age, the male offspring of the third generation were killed. The fatty acyl composition of isolated capillaries and choroid plexus was determined. The major changes noted in the fatty acid profile of isolated capillaries were a reduction (threefold) in the level of docosahexaenoic acid and, consequently, a fourfold increase in docosapentaenoic acid in sunflower oil-fed animals. The total percentage of polyunsaturated fatty acids was close to that in the soya oil-fed rats, but the ratio of n-3/n-6 fatty acids was reduced by threefold. In the choroid plexus, the C22:6n-3 content was also reduced, but by 2.6-fold, whereas the C22:5n-6 content was increased by 2.3-fold and the ratio of n-3/n-6 fatty acids was reduced by 2.4-fold. When the diet of sunflower oil-fed rats was replaced with a diet containing soya oil at 60 days of age, the recovery in content of n-6 and n-3 fatty acids started immediately after diet substitution; it progressed slowly to reach normal values after 2 months for C22:6n-5 and 2.5 months for C22:6n-3. The recovery in altered fatty acids of choroid plexus was also immediate and very fast. Recovery in content of C22:5n-6 and C22:6n-3 was complete by 46 days after diet substitution.     

Indonesian tapé ketan fermentation

Indonesian tapé ketan is a fermentation in which a mold, Amylomyces rouxii Calmette (Chlamydomucor oryzae Went and Prinsen Geerligs), in combination with one or more yeasts such as Endomycopsis burtonii converts steamed rice to a sweet-sour, slightly alcoholic paste. A study was made to determine the biochemical changes that occur in the substrate during fermentation. It was found that the product was ready for consumption after fermentation at 30 degrees C for 36 to 48 h. A. rouxii used about 30% of the total rice solids, resulting in a crude protein of 12% in 96 h, whereas the combination of the mold with E. burtonii reduced total solids by 50% in 192 h, causing crude protein to increase to 16.5%. Soluble solids increased from 5 to about 67% in 36 h and decreased to 12% at 192 h with A. rouxii alone, whereas soluble solids fell to about 8% at 192 h in the fermentation with both the mold and the yeast. The mold, by itself, reduced the starch content of the rice from 78 to 10% in 48 h and to less than 2% in 144 h. The mold plus yeast reduced the starch content to about 18% in 48 h; however the "starch" content did not fall below 6% even at 192 h, presumably because the yeast was producing glycogen, which was determined along with the residual starch. With both the mold and the mold plus yeast fermentations, reducing sugars increased from less than 1% to approximately 5% in 24 h and reached maximum concentration, 16 to 17%, between 36 and 48 h. A. rouxii by itself produced a maximum of about 5.6% (vol/vol) ethanol at 96 h. The highest concentration of ethanol (8%, vol/vol) was produced by the mold plus E. burtonii at 144 h. The mold by itself reduced the starting pH from 6.3 to about 4.0 in 48 h. The combination of the mold and yeast reduced the pH to 4.1 in 144 h. The mold increased total acidity to approximately 6.2 meq of H per 100 ml, and the combination of the mold and yeast increased the total acidity to 7.8 meq of H per 100 ml in 192 h. At 48 h there was practically no difference in the volatile acidity (0.20) for the combined fermentation compared with 0.26 meq of H per 100 ml for the mold fermentation. The mold and at least one species of yeast were required to develop the rich aroma and flavor of typical Indonesian tapé.     

盐度对哈氏仿对虾肌肉一般营养成分和氨基酸组成及含量的影响

通过调查不同盐度(12～36)环境下养殖的哈氏仿对虾(Parapenaeopsis hardwickii)肌肉一般营养成分和氨基酸组成及含量,研究了盐度对该虾肌肉营养品质的影响。结果显示,肌肉的水分随环境盐度升高出现显著的直线下降,而粗蛋白含量随环境盐度升高而直线升高;各盐度组的粗脂肪含量虽然没有明显差异,但其含量随盐度升高有一定的线性下降趋势;36盐度组的粗灰分含量比12、16、20和28盐度组的明显高,32盐度组的灰分含量比20和24盐度组的也明显高;所检测的肌肉干样16种氨基酸中,只有3种氨基酸的含量随盐度升高而升高,而其他13种氨基酸的含量随盐度升高明显降低,其中有10种氨基酸的含量在盐度12～24条件下的比盐度28～36条件下的明显高,而这些氨基酸含量在盐度12～24之间没有明显差异。各盐度组间氨基酸总量和鲜味氨基酸含量均没有明显差异;必需氨基酸和半必需氨基酸含量在盐度12～24条件下的均比盐度28～36条件下的明显高,而在12～24盐度组之间以及28～36盐度组之间均没有明显差异。必需氨基酸/氨基酸总量的比值和必需氨基酸/非必需氨基酸比值随盐度升高均明显线性降低;盐度12～24组的必需氨基酸指数(66.13～67.42)高于盐度28～36组的(62.56～64.46)。综上所述,盐度12～24环境下养殖的哈氏仿对虾肌肉营养价值相对较高,表现为低盐趋向,考虑到肌肉的水分含量和生长性能,哈氏仿对虾在养殖期间选择16～24盐度范围比较合适,同时也说明哈氏仿对虾适合大多数沿海地区环境的盐度条件。     

Indonesian Tapé Ketan Fermentation

Indonesian tapé ketan is a fermentation in which a mold, Amylomyces rouxii Calmette (Chlamydomucor oryzae Went and Prinsen Geerligs), in combination with one or more yeasts such as Endomycopsis burtonii converts steamed rice to a sweet-sour, slightly alcoholic paste. A study was made to determine the biochemical changes that occur in the substrate during fermentation. It was found that the product was ready for consumption after fermentation at 30°C for 36 to 48 h. A. rouxii used about 30% of the total rice solids, resulting in a crude protein of 12% in 96 h, whereas the combination of the mold with E. burtonii reduced total solids by 50% in 192 h, causing crude protein to increase to 16.5%. Soluble solids increased from 5 to about 67% in 36 h and decreased to 12% at 192 h with A. rouxii alone, whereas soluble solids fell to about 8% at 192 h in the fermentation with both the mold and the yeast. The mold, by itself, reduced the starch content of the rice from 78 to 10% in 48 h and to less than 2% in 144 h. The mold plus yeast reduced the starch content to about 18% in 48 h; however the “starch” content did not fall below 6% even at 192 h, presumably because the yeast was producing glycogen, which was determined along with the residual starch. With both the mold and the mold plus yeast fermentations, reducing sugars increased from less than 1% to approximately 5% in 24 h and reached maximum concentration, 16 to 17%, between 36 and 48 h. A. rouxii by itself produced a maximum of about 5.6% (vol/vol) ethanol at 96 h. The highest concentration of ethanol (8%, vol/vol) was produced by the mold plus E. burtonii at 144 h. The mold by itself reduced the starting pH from 6.3 to about 4.0 in 48 h. The combination of the mold and yeast reduced the pH to 4.1 in 144 h. The mold increased total acidity to approximately 6.2 meq of H⁺ per 100 ml, and the combination of the mold and yeast increased the total acidity to 7.8 meq of H⁺ per 100 ml in 192 h. At 48 h there was practically no difference in the volatile acidity (0.20) for the combined fermentation compared with 0.26 meq of H⁺ per 100 ml for the mold fermentation. The mold and at least one species of yeast were required to develop the rich aroma and flavor of typical Indonesian tapé.