Anti-stress and Anti-fatigue Effects of Fermented Rice Bran

The anti-stress and anti-fatigue effects on rats and mice of a hot water extract of rice bran fermented with Saccharomyces cerevisae IFO 2346 were investigated. Oral administration (1 g/kg/day) of the hot water extract of fermented rice bran (FRB) inhibited major changes in the weight of the adrenal, thymus, spleen and thyroid, showing an anti-stress effect. The hot water extract of FRB also inhibited increases in the GPT and LDH activity, cholesterol and glucose in the serum. The administration (1 g/kg/day) for 2 weeks significantly prolonged the swimming time, resulting in an increase in the anti-fatigue effect. It is considered from these results that FRB had anti-stress and anti-fatigue effects.     

Anti-stress and anti-fatigue effects of fermented rice bran.

The anti-stress and anti-fatigue effects on rats and mice of a hot water extract of rice bran fermented with Saccharomyces cerevisae IFO 2346 were investigated. Oral administration (1 g/kg/day) of the hot water extract of fermented rice bran (FRB) inhibited major changes in the weight of the adrenal, thymus, spleen and thyroid, showing an anti-stress effect. The hot water extract of FRB also inhibited increases in the GPT and LDH activity, cholesterol and glucose in the serum. The administration (1 g/kg/day) for 2 weeks significantly prolonged the swimming time, resulting in an increase in the anti-fatigue effect. It is considered from these results that FRB had anti-stress and anti-fatigue effects.     

Utilization of rice bran as nutrient source for fermentative lactic acid production

To reduce nutrient cost for lactic acid production, rice bran, one of agricultural wastes, was chosen as a nutrient source in this study. Although rice bran is rich in protein and vitamins, the use of rice bran without any treatment was inefficient in lactic acid production. Rice bran was treated by acid-hydrolysis before it was put in experiment, when it was hydrolyzed at initial pH 1, 30 g/L rice bran could provide a productivity to that degree of about 8 g/L YE, showing such a desirable result that the use of rice bran as nutrient source would be a solution for reducing nutrient cost. However, the addition of hydrolyzed rice bran prolonged lag phase of fermentation, especially, in the fermentation with rice bran hydrolyzed at initial pH 0.5, a prolonged lag phase of about 40 h was observed. According to the quantitative determination of thiamine, pyridoxine, organic nitrogen and carbon, the prolongation of lag phase might be the result from the destruction of B vitamins and excessive hydrolysis of protein. To shorten the lag phase, combining hydrolyzed rice bran with yeast extract (YE) of small amount was considered to be a solution. When 3g/L YE was combined with 30 g/L rice bran hydrolyzed at initial pH 1, obtained was a productivity 1.6 times higher than that of the control fermentation with 15 g/L YE.     

Gamma-linolenic acid production of Mucor rouxii by solid-state fermentation using agricultural by-products

Aims:  This study aims to maximize the yield of gamma-linolenic acid by a filamentous fungus, Mucor rouxii , using low cost production by solid-state fermentation.
Methods and Results:  We optimized substrate types and culture conditions including inoculum size and temperature. The optimal growth of M. rouxii was found in the cultures inoculated with 5 × 10⁵ spores g⁻¹ substrate. The fungal cells grew well on rice bran and soy bean meal, whereas a lower biomass was found in the cultures grown on polished rice, broken rice and spent malt grain. The GLA content was highly accumulated in rice bran ferment and its maximal content of about 6 g kg⁻¹ fermented mass was observed in the 5th-day culture grown at 30°C. However, the GLA content in the rice bran ferment was not enhanced by low temperature.
Conclusions:  The GLA production of M. rouxii could be enhanced by optimizing the agricultural by-product substrates and culture condition.
Significance and Impact of the Study:  Low cost GLA production process was achieved, and fermented product containing GLA can be incorporated into feed additives without further oil extraction to alternate the expensive plant oils.     

The content and distribution of steryl ferulates in wheat produced in Japan

Oryzanol contained in rice bran is a complex mixture of steryl ferulates (SFs) with many identified health benefits. Recently, SF has been shown to exist in other cereals such as wheat, rye, and corn. In this study, SFs in several wheats produced in Japan were analyzed. For instance, SF content of whole wheat grain, Yumekaori (Japan) was 15.2 ± 1.4 mg-oryzanol-equivalent/100 g grain, while that of the imported one, 1CW (Canada) was 11.4 ± 1.3 mg-oryzanol-equivalent/100 g grain. The main SF components in the examined wheats were campesteryl ferulate, campestanyl ferulate, and sitostanyl ferulate. SF distribution in whole wheat grain was investigated using 14 fractions produced by a conventional test milling machine. SF was intensively accumulated in the four bran fractions (24 ? 95 mg-oryzanol-equivalent/100 g bran fraction). These results suggest that the wheat bran would be an important source of SF.     

Administration of hot-water extract of brown seaweed Sargassum duplicatum via immersion and injection enhances the immune resistance of white shrimp Litopenaeus vannamei

The total haemocyte count (THC), phenoloxidase activity, and respiratory burst were examined when the white shrimp Litopenaeus vannamei (10.42+/-1.39 g) were immersed in seawater (34 per thousand) containing hot-water extract of brown alga Sargassum duplicatum at 100, 300 and 500 mg l(-1), or injected with hot-water extract of S. duplicatum at 2, 6, 10 and 20 microg g(-1). These parameters increased significantly when the shrimp were immersed in seawater containing hot-water extract at 300 and 500 mg l(-1) after 1 h, or when the shrimp were injected with hot-water extract at 10 and 20 microg g(-1) after 1 day. L. vannamei that were injected with hot-water extract at 6, 10 and 20 microg g(-1) had increased phagocytic activity and clearance efficiency to V. alginolyticus after 1-6 days. In another experiment, L. vannamei which had been immersed in seawater containing hot-water extract at 100, 300 and 500 mg l(-1), or injected with hot-water extract at 2, 6, 10 or 20 microg g(-1) were challenged with V. alginolyticus at 1 x 10(6), or 1.4 x 10(6) colony-forming units (cfu) shrimp(-1), and then placed in seawater. The survival of shrimp that received hot-water extract at either dose was significantly higher than that of control shrimp after 2 days, as well as at the termination of the experiment (6 days after the challenge). It is therefore concluded that L. vannamei that were immersed in hot-water extract of S. duplicatum at 300 mg l(-1), or the shrimp that were injected with hot-water extract at 10 microg g(-1) or less had increased immune ability as well as resistance to V. alginolyticus infection.     

Antioxidant,antibacterial, tyrosinase inhibitory,and biofilm inhibitory activities of fermented rice bran broth with effective microorganisms

The effective microorganism fermentation extract (EM-X) is a refreshment beverage widely consumed in East Asia. Due to the antioxidant property of the health beverage, EM-X has potential benefits for the human immune system and has been generally accepted in clinical practice. In search of new functions of EM-X, another version of EM-X, named EM-YU in this study, which is fermented from rice bran, seaweed, and kiwifruit with effective microorganisms demonstrates high antioxidant, antibacterial, tyrosinase inhibition activities, and biofilm inhibition activity of pathogenic bacteria. The antioxidant activity of 1.8% EM-YU was equivalent to that of 0.01 mg/mL vitamin C. Also, EM-YU clearly inhibited the cell growth of two pathogenic bacteria, Escherichia coli O157:H7 and Pseudomonas aeruginosa PAO1; and EM-YU broth (6.7%) inhibited (55 ± 1%) the activity of mushroom tyrosinase. Moreover, the ethyl acetate extract of EM-YU, called EM-YU-EX (0.6 mg/mL), 7-fold reduced the biofilm formation of E. coli O157:H7 without affecting its cell growth. It is the first report that EM-X variant possess antibacterial, tyrosinase inhibitory, and biofilm inhibitory activities. These results support other beneficial properties of the natural health product EM-X variant (EM-YU).     

The immunostimulatory effects of hot-water extract of Gelidium amansii via immersion, injection and dietary administrations on white shrimp Litopenaeus vannamei and its resistance against Vibrio alginolyticus

The total haemocyte count (THC), phenoloxidase activity, and respiratory burst were examined when white shrimp Litopenaeus vannamei were immersed in seawater (34 per thousand) containing hot-water extract of red alga Gelidium amansii at 200, 400 and 600 mg l(-1), injected with hot-water extract at 4 and 6 microg g(-1) shrimp, and fed diets containing hot-water extract at 0, 0.5, 1.0 and 2.0 g kg(-1). These parameters increased significantly when shrimp were immersed in seawater containing hot-water extract at 400 and 600 mg l(-1) after 1h, when shrimp were injected with hot-water extract at 6 microg g(-1) shrimp after one day, and when shrimp were fed diets containing hot-water extract at 1.0 and 2.0 g kg(-1) after 14 days. Phagocytic activity and clearance efficiency were significantly higher for the shrimp that were fed diets containing hot-water extract at 1.0 and 2.0 g kg(-1) than those of shrimp that were fed diets containing hot-water extract at 0 and 0.5 g kg(-1) after 14 and 28 days. In a separate experiment, L. vannamei which had received hot-water extract via injection, or fed diets containing hot-water extract, were challenged after 3h or 28 days with V. alginolyticus at 2 x 10(6) cfu shrimp(-1) and 1 x 10(6) cfu shrimp(-1), respectively, and then placed in seawater. The survival of shrimp that were injected with hot-water extract at 6 microg g(-1) was significantly higher than that of control shrimp after 1 day, and the survival of shrimp fed diets containing hot-water extract at 0.5, 1.0 and 2.0 g kg(-1) increased significantly after 3 days as well as at the end of the experiment (6 days after the challenge), respectively. It was concluded that L. vannamei that were immersed in hot-water extract at 400 mg l(-1), injected with hot-water extract at 6 microg g(-1) shrimp, and fed hot-water extract of G. amansii at 2.0 g kg(-1) or less showed increased immune ability as well as resistance to V. alginolyticus infection.     

Fermentative production of dl-lactic acid from amylase-treated rice and wheat brans hydrolyzate by a novel lactic acid bacterium, Lactobacillus sp.

Rice and wheat brans, without additional nutrients and hydrolyzed by alpha-amylase and amyloglucosidase, were fermented to DL-lactic acid using a newly isolated strain of Lactobacillus sp. RKY2. In batch fermentations at 36 degrees C and pH 6, the amount of lactic acid in fermentation broth reached 129 g l(-1) by supplementation of rice bran with whole rice flour. The maximum productivity was 3.1 g lactic acid l(-1) h(-1) in rice bran medium supplemented with whole rice flour or whole wheat flour.     

Production of cellulases and hemicellulases by Aspergillus niger KK2 from lignocellulosic biomass

To investigate the production of cellulases and hemicellulases from Aspergillus niger KK2, solid state fermentation (SSF) was performed by using different ratios of rice straw and wheat bran. When A. niger KK2 was grown on rice straw alone as a solid support in SSF, the maximum FPase activity was 19.5 IU g(-1) in 4 days. Also, CMCase (129 IU g(-1)), beta-glucosidase (100 IU g(-1)), xylanase (5070 IU g(-1)) and beta-xylosidase (193 IU g(-1)) activities were concurrently obtained after 5-6 days of fermentation. The higher enzyme activities produced by A. niger KK2 is a significant advantage from the viewpoint of practical saccharification reaction. Cellulases and hemicellulases produced by A. niger KK2 might be applied to pulp and paper industry, feed industry and chemical industry.     

Changes in lipid, fatty acids and phospholipids composition of whole rice bran after solid-state fungal fermentation

The aim of this study was to evaluate fermented rice bran phospholipids, lipids and fatty acid content in a fermentation solid system with Rhizopus oryzae fungus. For this, aliquots were withdrawn every 24h over 120 h. The content of phospholipids was determined by colorimetric method. Esterified fatty acids were separated by gas chromatography, then identified and quantified. The total lipids from fermented rice bran (FB) decreased from 20.4% to 11.2% in the range between 0 h and 120 h of fermentation while phospholipid contents were increased up to 2.4 mg P g(lipid)(-1). In fermented bran, oleic, palmitic and linoleic acids prevailed, with a decrease in saturated fatty acids (20%) and increase in the unsaturated ones (5%). This study showed that rice bran fermentation with R. oryzae can be applied to the production of phospholipids altering the saturated to unsaturated fatty acid ratio.     

Effect of organic complex compounds on Bacillus thermoamylovorans growth and glucose fermentation.

The effect of the concentration of a mixture (1/1 [wt/wt]) of yeast extract and bioTrypcase (YE+bT) on the growth and physiology of a new species, Bacillus thermoamylovorans, a moderately thermophilic, non-spore-forming, lactic acid-producing bacterium isolated from palm wine, was studied. At an initial glucose concentration of 100 mM, B. thermoamylovorans growth was limited when the concentration of YE+bT was lower than 5.0 g liter(-1); under these conditions, cellular yield reached a maximum value of 0.4 g of cells per g of YE+bT. Growth limitation due to deficiency in growth factors led to a significant shift in glucose metabolism towards lactate production. Lactate constituted 27.5 and 76% of the end products of glucose fermentation in media containing YE+bT at 20.0 and 1.0 g liter(-1), respectively. This result markedly differed from published data for lactic bacteria, which indicated that fermentative metabolism remained homolactic regardless of the concentration of YE. Our results showed that the ratio between cellular synthesis and energy production increased with the concentration of YE+bT in the culture medium. They indicate that the industrial production of lactic acid through glucose fermentation by B. thermoamylovorans can be optimized by using a medium where glucose is present in excess and the organic additives are limiting.     

Covalently attached ferulic acid in a proteoglycan from rice bran

A water-soluble proteoglycan, precipitated with ammonium sulfate from the hot-water extract of rice bran, contained ferulic acid, which was liberated by alkaline treatment. Evidence for the linkage between the carboxyl group of ferulic acid and the proteoglycan was obtained by the characterization of ferulic acid hydroxamate after treatment of the proteoglycan with hydroxylamine.     

Effect of yeast extract on Escherichia coli growth and acetic acid production

Fed batch cultures were performed to investigate the effect of yeast extract concentration on the kinetics of growth and acetic acid production of recombinant Escherichia coli BL21 in a synthetic medium. Three runs were performed with 40g/l total glucose concentration. The yeast extract/glucose ratio (YE/G; w/w), was 0.1, 0.05 and 0.025 in the feed. These decreasing YE/G values did not affect growth kinetics, but reduced the final cell concentration by about 10%, and also reduced the cell yield. Experiments with 60g/l total glucose concentration, one with a YE/G of 0.025 in the feed and the other without yeast extract, showed final acetic acid concentrations of 5.1 and 0.5g/l respectively, without any difference in cellular concentration. Although there was no significant influence on growth kinetics and final cellular concentration, the cell fermentative capacity was enhanced by yeast extract. The feed medium without yeast extract was the best condition for control purposes in high cell density cultures and for recombinant gene expression.     

Elicitor-induced rosmarinic acid accumulation and secondary metabolism enzyme activities in Solenostemon scutellarioides

This study aimed to improve rosmarinic acid (RA) production in the whole plant culture of Solenostemon scutellarioides through elicitation. Amongst selected elicitors methyl jasmonate (MJ), salicylic acid (SA), and yeast extract (YE) caused significant elevation in RA accumulation. Elicitation with MJ (50 μM) and SA (50 μM) caused almost 1.7 and 1.4-fold increase in RA accumulation, respectively, within day 1. While YE (100 μg ml^?1) elicitation showed highest RA content (~1.5-fold) in day 3. Preceding the elicitor-induced RA accumulation, there was a notable alteration in the specific activities of RA biosynthetic enzymes viz. phenylalanine ammonia lyase, tyrosine aminotransferase, hydroxyl-phenylpyruvate reductase and rosmarinic acid synthase up on MJ (50 μM), SA (50 μM) and YE (100 mg ml^?1) elicitation. Based on differential responses of aforementioned enzymes, RA synthesis was further scaled up through combination of elicitors in pre-optimized doses. In synergy study, at a time exposure with MJ + SA + YE and MJ + SA followed by YE after 24 h has been found to produce significant elevation of RA (2.0 and 1.9-fold, respectively) within 24 h while later maintained a steady state increased level (~1.7 ± 0.2-fold) over control up to day 7.     

Microbial status and rumen enzyme profile of crossbred calves fed on different microbial feed additives

AIMS: To test various microbial cultures as cattle feed additives. METHODS AND RESULTS: Four groups of newly born crossbred calves (average body weight 23.5 kg) were reared on green berseem and calf starter which was devoid of cereal grains. Milk was fed up to 8 weeks of age, starting with one tenths and gradually reducing to one twentieths of the body weight. One hundred millilitres of microbial feed additive or 100 g fermented feed was fed to the animals of group 2 (curd containing lactic acid bacteria at 10(8) cfu x ml(-1)), group 3 (Saccharomyces cerevisiae NCDC-49 at 10(6) cfu x ml(-1)) and group 4 (Lactobacillus acidophilus-15 at 10(8) cfu x ml(-1)). Group 1 served as control. The incidence and duration of diarrhoea was lower in the animals of probiotic fed groups as compared to control group. Out of three microbial feed additives, yeast feeding showed maximum suppression of diarrhoea followed by Lactobacillus and curd. CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: There was no effect of probiotic feeding on the log number of cells of lactic acid bacteria, yeast and coliform bacteria in the faeces and rumen liquor at any age. The activities of carboxymethylcellulase, xylanase, beta-glucosidase, alpha-glucosidase, alpha-amylase, protease, urease and pH of the rumen liquor remained unaffected by probiotic feeding at all ages tested in this experiment.     

Increasing α-linolenic acid content in rice bran by embryo-specific expression of ω3/Δ15-desaturase gene

Rice bran, the byproduct of brown rice, with a million tons produced annually, is the major storage organ for lipids. Increasing the α-linolenic acid (ALA) content in rice bran by biotechnology strategies is beneficial for both human health and the oil industries. We introduced two ω3/Δ15 fatty acid desaturase genes cloned from rice and soybean into rice under the control of an embryo-specific promoter, REG. The ALA content (dry weight) in embryos and bran was increased up to 6.09 and 5.86 mg/g, respectively, in transgenic lines, which was 25.4- and 27.9-fold higher than the 0.24 and 0.21 mg/g in the nontransformant control. ALA accounted for 46.7 and 44.3 % of the total fatty acids in embryos and bran, respectively, of transgenic plants, which was comparable to that in linseed and perilla seeds. The trait of high ALA content was stably inherited. The enhanced ALA content was preferentially located at the sn-2 position in triacylglycerols, which are digestible and absorbable for humans. ALA-enriched rice bran may help alleviate human health problems caused by ALA deficiency, with the production of healthy bran oil for humans and feed for animals.     

鲍姆纤孔菌（桑黄）不同方式发酵的菌丝体生物活性比较

利用液体发酵、木屑固体发酵和米饭固体发酵3种方式培养鲍姆纤孔菌（桑黄）菌丝体,对菌丝体醇提物的体外抗氧化、抗肿瘤和抗衰老生物活性进行了研究。结果表明,木屑固体发酵、液体发酵和米饭固体发酵的菌丝体醇提物清除H2O2自由基的IC50值分别为78.28±0.32、27.73±0.57和7.84±0.37;米饭培养的桑黄菌丝体醇提物在低浓度500μg/mL下对超氧阴离子自由基清除作用到达80%,在相同的浓度下对DPPH自由基清除率也明显高于其他两种方法,表现出较高的抗氧化活性。木屑以及米饭培养方法得到的菌丝体对PC12神经细胞损伤修复均有较好的效果,液体培养的桑黄菌丝体表现的修复作用较低;液体发酵培养的菌丝体醇提物浓度在100μg/mL时,对肿瘤细胞HepG2的抑制率达70%,高于其他两种培养方法的抑制作用。     

Response surface methodology for the optimization of <Emphasis Type="Italic">α</Emphasis>-amylase production by <Emphasis Type="Italic">Streptomyces</Emphasis> sp. ML12 using agricultural byproducts

Amylases constitute one of the most important groups of enzymes for commercial use. In the present study, production of α-amylase was optimized using a newly isolated actinobacterial strain from the coral reef environment of the Gulf of Mannar Biosphere Reserve, India. It was identified as Streptomyces sp. ML12 based on chemotaxonomy, cultural and morphological characteristics, carbon source utilization and 16S rRNA gene sequencing. Fermentation variables were selected in accordance with the Plackett-Burman design and were optimized by response surface methodology. Five significant variables (rice bran and wheat bran — both agricultural byproducts, sodium chloride, magnesium sulphate and incubation period) were selected for the optimization via central composite design. The optimal features were rice bran (5.5 g/100 mL), wheat bran (5.3 g/100 mL), sodium chloride (2.8 g/100 mL), magnesium sulphate (1.4 g/100 mL) and 8 days of incubation period. Optimization of the medium with the above tested features increased the amylase yield by 4.4-fold.     

Ultrahigh diterpenoid tanshinone production through repeated osmotic stress and elicitor stimulation in fed-batch culture of <Emphasis Type="Italic">Salvia miltiorrhiza</Emphasis> hairy roots

Hyperosmotic stress (OS, created with 50 g/L sorbitol) and a yeast elicitor (YE, polysaccharide fraction of yeast extract) applied to Salvia miltiorrhiza hairy root cultures had a synergistic effect on the diterpenoid tanshinone production. With a single OS+YE treatment and nutrient feeding, the total tanshinone content of roots was increased by sevenfold (from 0.2 to 1.6 mg/g dry weight (dw)) and the volumetric yield by 13-fold (from 1.95 to 27.4 mg/L) compared to the batch control culture. With repeated feeding of OS and nutrient medium in an extended fed-batch culture process (i.e., 10 mL fresh medium with 50 g/L sorbitol 25 mg/L YE, every 5 days from day 21 to day 60), the total tanshinone content of roots was increased to 18.1 mg/g dw (or 1.8 wt.%) and the volumetric tanshinone yield to 145 mg/L, which were about 100-fold and 70-fold of those, respectively, in the batch control. Another interesting finding was the presence of root fragments (fine particles) with extremely high tanshinone content in the OS+YE treated cultures. It was also possible to reuse the sorbitol medium for the hairy root growth and tanshinone production to reduce the medium expenses.