Value-added subcritical water hydrolysate from rice bran and soybean meal

New value-added product was derived from agricultural by-products: rice bran and soybean meal by means of subcritical water (SW) hydrolysis. The effect of temperature (200-220 degrees C), reaction time (10-30 min), raw material-to-water weight ratio (1:5 and 2:5), was determined on the yields of protein, total amino acids, and reducing sugars in the soluble products. The suitable hydrolysis time was 30 min and the proper weight ratio of the raw material-to-water was 1:5. The reaction temperature suitable for the production of protein and amino acids was 220 degrees C for raw and deoiled rice bran, 210 degrees C for raw soybean meal, and 200 degrees C for deoiled soybean meal. The products were also found to have antioxidant activity as tested by ABTS(.)(+) scavenging assay. In addition, sensory evaluation of milk added with the hydrolysis product of deoiled rice bran indicated the potential use of the product as a nutritious drink.     

Changes in moisture content,mycoflora and aflatoxin content of rice bran during storage

The changes in moisture content, storage mycoflora and aflatoxin B₁ (AFB₁) in bran from untreated or raw rice (Rr) and parboiled rice (Pbr) stored in small lots in polyethylene bags were studied at 15-day intervals up to 60 days, using five lots of each type of bran. Deterioration was more rapid with reference to all the three parameters, in Rr bran compared to Pbr bran, the former becoming completely overgrown and caked with fungi by the end of 60 days.Aspergillus flavus was the dominant fungus in Pbr bran, whereasA. candidus andTrichoderma viride were abundant in Rr bran. The frequency of incidence as well as concentration of AFB₁ increased with storage time in both types of bran, but the rate of increase as well as overall concentration were much higher in Rr bran. Thus raw rice bran is unsuitable for prolonged storage.Abbreviations AFB₁ aflatoxin B₁ - MC moisture content - Pbr parboiled rice - Rr raw rice     

Effects of concentrated carbon dioxide on the fruiting of several cultivated basidiomycetes (II)

Edible basidiomycetesFlammulia velutipes andPleurotus ostreatus were cultivated in the usual manner on media based on sawdust and rice bran, and the cultures were exposed to slowly flowing CO₂-enriched air (550 (control), 3,000, 6,000, and 9,000µl/l) for seven days at different stages of cultivation. When the cultures were exposed at the primordium stage (less than 10 mm in length), length and yield of fruit-bodies increased and pileus expansion was slightly inhibited inF. velutipes, while inP. ostreatus length increased, yield decreased, and pileus expansion was greatly inhibited. When the cultures with fruit-bodies larger than 10 mm were exposed, length and yield were insensitive and pileus expansion was greatly inhibited inF. velutipes, while inP. ostreatus length was insensitive, but pileus expansion was heavily damaged by trumpet-like deformation and yield decreased. The different action of CO₂ on the two species appeared to be due to the different anatomical structures of their fruit-bodies.     

Effect of culture substrates on virulence of Beauveria bassiana (Ascomycota: Cordycipitaceae) conidia against the browntail moth,Euproctis chrysorrhoea (Lepidoptera: Lymantriidae)

We studied the effect of different solid substrates on virulence of two Beauveria bassiana isolates against the browntail moth, Euproctis chrysorrhoea (L.) (Lep.: Lymantriidae). Conidia produced on wheat grains, wheat flour, wheat bran, rice flour, rice bran, rice paddy, corn flour, millet, and Sabouraud's dextrose agar with 1% yeast extract (SDAY) as control were compared. There were significant differences among these substrates for their effects on the virulence of produced conidia. Applying 10⁷ conidia/mL of B. bassiana EUT105, produced on rice bran caused the highest (84.9%) and on rice flour, the lowest (57.6%) mortalities. Bioassay on fifth-instar larvae using aerial conidia harvested from wheat grains, rice paddy, and SDAY indicated that conidia from wheat grains had the highest virulence while those from rice paddy, the lowest.     

Extraction of protein and amino acids from deoiled rice bran by subcritical water hydrolysis

This study investigated the production of value-added protein and amino acids from deoiled rice bran by hydrolysis in subcritical water (SW) in the temperature range between 100 and 220 degrees C for 0-30 min. The results suggested that SW could effectively be used to hydrolyze deoiled rice bran to produce useful protein and amino acids. The amount of protein and amino acids produced are higher than those obtained by conventional alkali hydrolysis. The yields generally increased with increased temperature and hydrolysis time. However, thermal degradation of the product was observed when hydrolysis was carried out at higher temperature for extended period of time. The highest yield of protein and amino acids were 219 +/- 26 and 8.0 +/- 1.6 mg/g of dry bran, and were obtained at 200 degrees C at hydrolysis time of 30 min. Moreover, the product obtained at 200 degrees C after 30 min of hydrolysis exhibited high antioxidant activity and was shown to be suitable for use as culture medium for yeast growth.     

Cultivation characteristics ofIsaria japonica

Cultivation characteristics of fruit-body (synnema) formation ofIsaria japonica were examined using liquid and solid media in order to produce fruit-bodies on a large scale. Mycelia grew well at 18–28°C on PDA medium with an initial pH of 7.0. The formation of fruit-bodies ofI. japonica was induced by lowering temperature to below 20°C in PD liquid medium. In sawdust-rice bran basal medium mixed with pupal powder prepared from silkworms (Bombyx mori), the fresh weight of fruit-bodies increased with increasing content of pupal powder. The highest yields of fruit-bodies were obtained in carbon-rich barley grain medium supplemented with pupal powder. The fruit-bodies grown under CO₂ concentrations of 1,000 μl/L had coral-like, many-branched synnemata with numerous conidiospores, whereas those formed under high concentrations (9,000 μl/L) of CO₂ had unbranched and longer synnemata. High concentrations of CO₂ remarkably inhibited conidiospore formation on synnemata. Continuous high-intensity illumination at 2.93 W·m⁻² inhibited the elongation of synnemata, and low-intensity illumination at 0.088 W·m⁻² slightly inhibited the branching of synnemata. Fruit-bodies were produced on the pupa metamorphosed from living larvae ofAgrotis fucosa placed on the surface of a culture ofI. japonica incubated in sawdust-rice bran medium.     

Intensification of Fructosyltransferases and Fructo-Oligosaccharides Production in Solid State Fermentation by Aspergillus awamori GHRTS

The present work was aimed to investigate the impact of the solid substrates mixture on Fructosyltransferases (FTase) and Fructo-oligosaccharides (FOS) production. An augmented simplex lattice design was used to optimize a three component mixture for FTase production. Among selected substrates corn cobs has highest impact on FTase production followed by wheat bran and rice bran. All two substrates and three substrate combinations showed the highest enzyme production than their individual levels. Among the tested various models quadratic model was found to be the best suitable model to explain mixture design. Corncobs, wheat bran and rice bran in a ratio of approximately 45:29:26 is best suitable for the FTase production by isolated Aspergillus awamori GHRTS. This study signifies mixture design could be effective utilize for selection of best combination of multi substrate for improved production of high value products under solid state fermentation.     

Production of multifunctional lipases by Penicillium verrucosum and Penicillium brevicompactum under solid state fermentation of babassu cake and castor meal

The main objective of this work was to optimize lipase production, in terms of hydrolytic and esterification activities, by Penicillium brevicompactum and Penicillium verrucosum in solid state fermentation using agroindustrial residues as raw material. Maxima hydrolytic activities of 48.6 and 87.7 U/g were achieved when P. brevicompactum was cultured in babassu cake and castor meal, respectively. Higher esterification activities (around 244 U/g) were achieved when P. brevicompactum was used as microorganism and babassu cake as raw material. Different experimental conditions led to these promising values, clearly showing that no correlation can be attributed between hydrolytic and esterification activities. In spite of the several applications of lipases which are capable of catalyze synthesis reactions, only few works in this subject are presented in the literature, especially when low cost raw materials are used.     

Production of (R)-2-(4-hydroxyphenoxy) propionic acid by Beauveria bassiana ZJB16007 in solid state fermentation using rice bran

Abstract

R-2-(4-hydroxyphenoxy)propionic acid (R-HPPA) is a key chiral intermediate for phenoxypropionic acid herbicide synthesis. In this study, to improve the production of R-HPPA with B. bassiana ZJB16007, the cultivation conditions in solid-state fermentation (SSF) were investigated. The effects of various substrates on R-HPPA production were evaluated and the process parameters were also optimized. The results showed that rice bran was the optimal substrate for R-HPPA production. The optimal medium components and cultivation conditions were: rice bran: silkworm chrysalis powder = 5.25: 2.25 (g: g), nutrient salts solution 12?mL which contained 50?g/L R-PPA, pH 5.0, and cultivated at 28?°C for 11 days. Under the optimized conditions, the transformation of R-HPPA was significantly improved and the yield of R-HPPA reached 77.78%, which was 15.14% higher than that of the control (67.55%). Therefore, SSF may serve as an alternative for R-HPPA production by B. bassiana ZJB16007.     

Effects of prefermented cereal-derived substrates (ground barley and rye bran) enriched with fungal γ-linolenic acid on rumen fermentation parameters and lipid metabolism in vitro

Aims: To increase rumen output of γ‐linolenic acid (GLA), we used two cereal‐derived substrates, ground barley (GB) and rye bran (RB), enriched with fungal GLA as components of feed rations. We examined their effects on rumen fermentation patterns, lipid metabolism and the ciliated protozoan population in an artificial rumen. Methods and Results: Four diets consisting of meadow hay (MH) plus unfermented (GB or RB) or prefermented (GB ? TE or RB ? TE) cereal‐derived substrates were fermented in an artificial rumen with ovine rumen inoculum. The cereal‐derived substrates were prefermented with the fungus Thamnidium elegans (TE) by fungal solid‐state fermentation. The diets with TE increased the rumen input of dietary GLA (mg day^?1) from 0 to 21 (GB ? TE) or 26 (RB ? TE). Both experimental diets increased the rumen output of GLA (P < 0·001). Adverse effects on the ciliate population were observed. Both diets also had an effect on the fatty acids profile. Fermentation patterns were also affected with MH + RB ? TE. Conclusion: Cereal‐derived substrates enriched with GLA effectively enhanced the output of GLA in artificial rumen. Significance and Impact of the Study: The ability of the fungal strain T. elegans to grow and utilize various agro‐industrial substrates might be useful in developing potential new animal diets enriched in GLA.     

Evaluation of shelf life and rock phosphate solubilization of Burkholderia sp. in nutrient-amended clay, rice bran and rock phosphate-based granular formulation

Five phosphate-solubilizing bacteria (PSB) used in this study were isolated based on their ability to solubilize tricalcium phosphate (TCP) in Pikovskaya’s medium. Among the tested bacterial strains Burkholderia sp. strain CBPB-HIM showed the highest solubilization (363 μg of soluble P ml⁻¹) activity at 48 h of incubation. Further, this strain has been selected to assess its shelf life in nutrient-amended and -unamended clay, rice bran and rock phosphate (RP) pellet-based granular formulation. The results showed that the maximum viability of bacterium was observed in clay and rice bran (1:1) + 10% RP pellets than clay-RP pellets, irrespective of tested storage temperatures. Further, clay and rice bran (1:1) + 10% RP pellets amended with 1% glucose supported the higher number of cells compared to glycerol-amended and nutrient-unamended pellets. In this carrier solubilization of Morocco rock phosphate (MRP) by Burkholderia sp. strain CBPB-HIM was also investigated. The maximum of water and bicarbonate extractable P (206 and 245 μg P g⁻¹ of pellet respectively) was recorded in clay and rice bran (1:1) + 10% RP pellets amended with 1% glucose and glycerol respectively on day 5 of incubation. Therefore, this study proved the possibility of developing granular inoculant technology combining clay, rice bran and RP as substrates with phosphate-solubilizing Burkholderia.     

Protein enrichment and digestibility of soft rush (<Emphasis Type="Italic">Juncus effusus</Emphasis>) and rice residues using edible mushrooms <Emphasis Type="Italic">Pleurotus ostreatus</Emphasis> and <Emphasis Type="Italic">Pleurotus sajor-caju</Emphasis>

Pleurotus species are found to be among the most efficient lignocellulolytic types of white-rot fungi. Rice is the main grain cultivated in the extreme south of Brazil. Defatted rice bran and straw are by-products of low aggregate value. Soft rush (Juncus effusus) is a common native plant also very abundant in the region. In the present work, we evaluated changes in substrate composition after growth of two white-rot fungal species: Pleurotus ostreatus and Pleurotus sajor-caju, aiming to increase protein content and digestibility from substrates through solid fermentations and obtain edible mushrooms of high aggregate value. For that, defatted rice bran, defatted rice straw and soft rush were utilized as substrate. The influence of the variables thermal treatment temperature of substrate, substrate moisture and concentration were evaluated on the protein content, digestibility and biological efficiency. The highest protein enrichment of rice bran in P. sajor-caju-fermented medium was due the fact that there was no fructification in these media, while for the P. ostreatus-fermented medium, part of the synthesized protein was converted into mushrooms. The highest protein enrichments were verified in medium with 80% moisture and 25% soft rush (47.1% using P. ostreatus and 49.0% using P. sajor-caju). A higher digestible protein increase was obtained for both species in media with 70% moisture and 25% soft rush.     

Natural occurrence of toxigenic fungi and mycotoxins in rice bran

Thirty four samples of rice bran, of which 9 were from raw (untreated) rice (RR) and 25 from parboiled rice (PbR) were collected from commercial rice mills in and around Madras and analysed for storage mycoflora and mycotoxins. Fungi of the Aspergillus flavus group were present in 29 of the 34 samples (8 from RR and 21 from PbR) in quantities ranging from <1–432 thousand propagules/g, though not always as the dominant mycoflora. Fungal numbers were usually higher in RR than in PbR samples.Five of the 9 RR samples and 6 of the 25 PbR samples were positive for aflatoxins. Among 29 isolates of A. flavus obtained from the bran samples, 16 isolates –6 from RR bran and 10 from PbR bran — were found to be toxigenic in vitro. Some isolates of A. candidus also seemed to produce aflatoxin and other fluorescent substances.     

Cellulolytic enzymes on lignocellulosic substrates in solid state fermentation by <Emphasis Type="Italic">Aspergillus niger</Emphasis>

The production of cellulolytic enzymes by Aspergillus niger on lignocellulosic substrates groundnut fodder, wheat bran, rice bran and sawdust in solid state fermentation in a laboratory scale was compared. Czapek Dox liquid broth amended with cellulose (0.5%) was used to moisten lignocellulosic solid supports for cultivation of Aspergillus niger. The production of filter paperase, carboxymethyl cellulase and -glucosidase were monitored at daily intervals for 5 days. The peak production of the enzymes occurred within 3 days of incubation. Among solid supports used in the study, wheat bran was the best solid matrix followed by groundnut fodder in production of cellulolytic enzymes in solid state fermentation. Groundnut fodder supported significant production of FPase (2.09 FPU/g), CMCase (1.36 U/g) and -glucosidase activity (0.0117 U/g) in solid state fermentation. Considerable secretion of protein (5.10 mg/g) on groundnut fodder at peak time interval 1st day of incubation was recorded.     

Responses of Hebeloma radicosum fruit-bodies to light and gravity: negatively gravitropic and nonphototropic growth

Responses of the long-rooting agaric Hebeloma radicosum fruit-bodies to light and gravity were studied. In light from below or obliquely below, fruit-bodies grew straight downward with gills tilted and cap swollen and waved if they had emerged downward from the culture medium, or bent upward from the beginning if they had emerged obliquely downward. In light from above or obliquely above, they grew upward if they had emerged upward. Thus, they did not grow toward unilateral light from obliquely below or obliquely above, and hence their growth was nonphototropic and negatively gravitropic from the beginning of development. Even the straight downward growth seems to be latently negative-gravitropic. In the dark, fruit-bodies grew upward, forming pseudorhizas, but they remained immature; they matured only in the light. These characteristics may be related to the growth habits of the fungus colonizing deep in the ground, forming primordia there, and developing mature fruit-bodies on the ground. Received: March 26, 2001 / Accepted: July 12, 2001     

Enhanced production of raw starch degrading enzyme using agro-industrial waste mixtures by thermotolerant Rhizopus microsporus for raw cassava chip saccharification in ethanol production

In the present study, solid-state fermentation for the production of raw starch degrading enzyme was investigated by thermotolerant Rhizopus microsporus TISTR 3531 using a combination of agro-industrial wastes as substrates. The obtained crude enzyme was applied for hydrolysis of raw cassava starch and chips at low temperature and subjected to nonsterile ethanol production using raw cassava chips. The agro-industrial waste ratio was optimized using a simplex axial mixture design. The results showed that the substrate mixture consisting of rice bran:corncob:cassava bagasse at 8?g:10?g:2?g yielded the highest enzyme production of 201.6?U/g dry solid. The optimized condition for solid-state fermentation was found as 65% initial moisture content, 35°C, initial pH of 6.0, and 5?×?10⁶ spores/mL inoculum, which gave the highest enzyme activity of 389.5?U/g dry solid. The enzyme showed high efficiency on saccharification of raw cassava starch and chips with synergistic activities of commercial α-amylase at 50°C, which promotes low-temperature bioethanol production. A high ethanol concentration of 102.2?g/L with 78% fermentation efficiency was achieved from modified simultaneous saccharification and fermentation using cofermentation of the enzymatic hydrolysate of 300?g raw cassava chips/L with cane molasses.     

Transmission by Laodelphax striatellus Fallen of Rice black‐streaked dwarf virus from Frozen Infected Rice Leaves to Healthy Plants of Rice and Maize

Rice black‐streaked dwarf virus (RBSDV) is transmitted naturally to important crops such as rice, maize, barley and wheat in a persistent manner by the planthoppers, Laodelphax striatellus, Unkanodes sapporona and Unkanodes albifascia. Insect vector transmission tests are the basis for identifying viral incidence, evaluating the resistance of varieties and selecting resistance sources for rice and maize breeding. A simple, rapid and reliable method is described by which virus‐free small brown planthoppers (L. striatellus) acquired RBSDV from frozen infected rice leaves and transmitted it to healthy rice and maize plants. After feeding on frozen infected rice leaves, the planthoppers were tested by RT‐PCR for the presence of virus after 10, 15, and 22 days, respectively. The percentages of RBSDV‐containing insects were 0, 25 and 71.43% of L. striatellus fed on frozen infected rice leaves compared to 0, 28.25 and 71.43% of L. striatellus fed on fresh infected rice leaves, respectively. In transmission tests, three of eight rice seedlings (37.5%) and four of eight maize seedlings (50%) were inoculated by the planthoppers that had fed previously on frozen leaves and had allowed a 22 days latent period and showed typical disease symptoms. As a positive control, four of eight rice seedlings (50%) and four of six maize seedlings (66.67%) became infected. All rice and maize plants expressing disease symptoms were identified as virus‐positive by RT‐PCR. These results indicated that the planthoppers acquired RBSDV from frozen infected leaves and transmitted the virus to healthy plants.     

Fractional Extraction of Rice Bran Oil with Supercritical Carbon Dioxide

Extraction of oil from rice bran with supercritical carbon dioxide (SC-C0₂) at pressures of 150 to 350 kg/cm² G at 40°C was demonstrated. The constituents of the selective fractions obtained at different pressures differed. Fractions obtained at higher pressures contained less free fatty acid and waxes or unsaponifiables. The phosphorus and iron contents were very low in the SC-C0₂ extracted oil, while the color of the oil was significantly lighter than that of hexane-extracted oil. The yield of low acid value oil was comparable to that for hexane extraction. One problem of the SC-C0₂ oil is its poor oxidation stability. This method of extraction may be effective in simplifying the processing of edible rice bran oil. Grinding the raw material was found to be effective in decreasing the C0₂ required and shortening the extraction period.     

Ergosterol as an indicator of mould growth on wood in relation to culture age, humidity stress and nutrient level

Changes in ergosterol content in cultures of Penicillium brevicompactum and Aspergillus versicolor on wood with time, changes in humidity or addition of glucose solutions to wood were studied with HPLC. Lowering of the humidity level caused a very large decline in ergosterol content of cultures of P. brevicompactum on wood over a 10 day period, although small amounts remained after this time. After an initial increase, up to an inoculation time of 45 days, reductions were also observed in control samples maintained at 100% RH, but these were smaller. The amount of ergosterol decreased to very low levels in wood impregnated with low levels of glucose during a 93 day incubation period. Ergosterol concentration in hyphae produced in surface liquid cultures was shown to be higher in mycelia growing on media enriched with nitrogen or with more available nutrients. The concentration of ergosterol in the mycelia of P. brevicompactum in surface liquid cultures varied by a factor of 5 from 2 to 10 mg g⁻. The results clearly show that ergosterol present in solid materials in mainly related to active biomass. With certain prerequisites, ergosterol determinations could also be used for total fungal biomass estimations on wood.     

The life history parameters of Diaphanosoma excisum (Cladocera), grown in different culturing media

Life history parameters of Diaphanosoma excisum (SARS) were derived from culture tube experiments with media comprising mohua oil cake, mustard oil cake, cow dung and rice bran. Body length, longevity, fecundity and progeny production were highest in rice bran medium. First reproduction time (A) in different media ranged from 2 to 6 days, while the reproduction peak (T) varied from 4 to 13 days. The total offspring per female (S) was directly dependent upon the mean number of eggs per female per life span. The intrinsic (r) and finite rate (e^r) of natural increase were larger for rice bran than for other media.