Detection, isolation, and characterization of a novel impurity from several folic acid products / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Folic acid belongs to the group of water-soluble B vitamins and naturally exists in multiple forms in a wide variety of foods such as legumes, vegetables, liver, and milk (Iyer and Tomar, 2009; Lyon et al., 2020). It is involved in many biochemical reactions critical for cell division, such as purine and pyrimidine biosynthesis, DNA/RNA biosynthesis, and amino acid metabolism (Iyer and Tomar, 2009). Mammals cannot synthesize folic acid and thus they must acquire it from food. Although folic acid is ubiquitous in foods, folic acid deficiency still often occurs due to various causes such as unhealthy diet (Hildebrand et al., 2021; Iimura et al., 2022), disease-related malabsorption (Arcot and Shrestha, 2005), medication-related depletion (Arcot and Shrestha, 2005), or vitamin B12 deficiency (Fishman et al., 2000). Folic acid deficiency has been associated with several health problems, such as anemia (Carmel, 2005; Bailey and Caudill, 2012), cancer (Duthie, 1999), cardiovascular diseases (Wald et al., 2002), neural tube defects in newborns (van der Put et al., 2001), neuropsychiatric dysfunction (Shea et al., 2002), depression (Falade et al., 2021), inflammatory diseases (Suzuki and Kunisawa, 2015; Jones et al., 2019), and eye diseases (Sijilmassi, 2019). To prevent folic acid deficiency, its daily intake (400 μg/d) has been recommended for adults in the European Union, and its increased intake (600 μg/d) is advised for women before and during pregnancy (FAO/WHO, 2002; IOM, 2004). The New Zealand government mandated the fortification of non-organic wheat flour with folic acid in July 2021, and the UK government mandated the fortification of non-wholemeal wheat flour with folic acid in September 2021 (Haggarty, 2021).

Effect of wheat-grain moxibustion on the expression of 5-HT and cortisol in the serum, and MR and GR in the hippocampus in rats with hypothyroidism complicated with depression / 中国针灸

OBJECTIVE@#To observe the effect of wheat-grain moxibustion on behavior, 5-hydroxytryptamine (5-HT) and cortisol in the serum, mineralocorticoid receptor (MR) and glucocorticoid receptor (GR) in the hippocampus in rats with hypothyroidism complicated with depression, and to explore the possible mechanism of wheat-grain moxibustion on improving depression in rats with hypothyroidism.@*METHODS@#A total of 32 SPF SD rats were randomly divided into a blank group, a model group, a medication group and a wheat-grain moxibustion group, 8 rats in each group. Except for the blank group, the rats in the remaining groups were treated with intragastric administration of 0.1% propylthiouracil (PTU) suspension at 1 mL/100 g, once a day for 4 weeks to establish the rat model of hypothyroidism, and whether the rats were accompanied with depression-like behavior determined through behavioristics evaluation. The rats in the medication group were intervened with euthyrox at 0.9 mL/100 g, once a day, for 4 weeks; the rats in the wheat-grain moxibustion group were treated with wheat-grain moxibustion at "Dazhui" (GV 14), "Mingmen" (GV 4), "Shenshu" (BL 23) and "Pishu" (BL 20), 7 cones each acupoint, once a day, six times a week for 4 weeks. After the intervention, the depression status was observed by behavioristics test; the contents of thyroid stimulating hormone (TSH), total thyroxine (TT4), 5-HT and cortisol in the serum were detected by ELISA; the protein expressions of MR and GR in hippocampus were detected by Western blot; the expressions of MR mRNA and GR mRNA in the hippocampus were detected by real-time PCR.@*RESULTS@#Before the intervention, compared with the blank group, the scores of open field test (OFT) were decreased and the immobility time of tail suspension test (TST) was prolonged (P<0.05); the serum TSH contents were increased and TT4 contents were decreased (P<0.01) in the other three groups. After the intervention, compared with the model group, the vertical score of OFT was increased and the immobility time of forced swimming test (FST) was prolonged in the medication group (P<0.05), while the scores of three items of OFT were increased (P<0.05, P<0.01), and the immobility time of FST and TST was shortened in the wheat-grain moxibustion group (P<0.01, P<0.05). Compared with the medication group, the immobility time of TST and FST in the wheat-grain moxibustion group was shorter (P<0.05, P<0.01). Compared with the blank group, in the model group, the contents of serum TSH and cortisol were increased (P<0.01, P<0.001), while the contents of serum TT4 and 5-HT were decreased (P<0.01, P<0.001). Compared with the model group, the contents of serum TT4 and 5-HT were increased, while the contents of serum TSH and cortisol were decreased in the medication group and wheat-grain moxibustion group (P<0.01, P<0.05). Compared with the blank group, the protein and mRNA expression of MR, GR in the hippocampus in the model group was decreased (P<0.01, P<0.05, P<0.001); compared with the model group, the protein and mRNA expression of MR in the hippocampus in the medication group were increased (P<0.05), and the protein expression of MR, GR and mRNA expression of MR in the hippocampus in the wheat-grain moxibustion group were increased (P<0.05, P<0.01). Compared with the medication group, the expression of MR mRNA in the wheat-grain moxibustion group was increased (P<0.05).@*CONCLUSION@#Wheat-grain moxibustion could significantly improve thyroid function and depression in rats with hypothyroidism. Its mechanism may be related to up-regulating the protein and mRNA expression of MR and GR in the hippocampus, and then affecting the expression of serum cortisol and 5-HT.

Production of wheat-doubled haploids resistant to eyespot supported by marker-assisted selection

BACKGROUND: Wheat is one of the most important crops cultivated all over the world. New high-yielding cultivars that are more resistant to fungal diseases have been permanently developed. The present study aimed at the possibility of accelerating the process of breeding new cultivars, resistant to eyespot, by using doubled haploids (DH) system supported by marker-assisted selection. RESULTS: Two highly resistant breeding lines (KBP 0916 and KBH 4942/05) carrying Pch1 gene were crossed with the elite wheat genotypes. Hybrid plants of early generations were analyzed using endopeptidase EpD1 and two SSR markers linked to the Pch1 locus. Selected homozygous and heterozygous genotypes for the Pch1-linked EpD1b allele were used to produce haploid plants. Molecular analyses were performed on haploids to identify plants possessing Pch1 gene. Chromosome doubling was performed only on haploid plants with Pch1 gene. Finally, 65 DH lines carrying eyespot resistance gene Pch1 and 30 lines without this gene were chosen for the eyespot resistance phenotyping in a field experiment. CONCLUSIONS: Results of the experiment confirmed higher resistance to eyespot of the genotypes with Pch1 in comparison to those without this gene. This indicates the efficiency of selection at the haploid level.

Impact of phosphate solubilizing bacteria on wheat (Triticum aestivum ) in the presence of pesticides / Impacto das bactérias solubilizantes de fosfato no trigo (Triticum aestivum ) na presença de pesticidas

Abstract Three phosphate solubilizing bacteria were isolated and identified by 16S rRNA sequencing as Pseudomonas putida, Pseudomonas sp and Pseudomonas fulva . The strains were subjected to plant biochemical testing and all the PGPR attributes were checked in the presence of pesticides (chlorpyrifos and pyriproxyfen). The phosphate solubilizing index of strain Ros2 was highest in NBRIP medium i.e 2.23 mm. All the strains showed acidic pH (ranges from 2.5-5) on both medium i.e PVK and NBRIP. Strain Ros2 was highly positive for ammonia production as well as siderophore production while strain Rad2 was positive for HCN production. The results obtained by the strains Rad1, Rad2 and Ros2 for auxin production were 33.1, 30.67 and 15.38 µg ml-1, respectively. Strain Rad1 showed 16% increase in percentage germination in comparison to control in the presence of pesticide stress. Most promising results for chlorophyll content estimation were obtained in the presence of carotenoids upto 6 mgg-1 without stress by both strains Rad1 and Rad2. Study suggests that especially strain Ros2 can enhance plant growth parameters in the pesticide stress.

Resumo Três bactérias solubilizantes de fosfato foram isoladas e identificadas por seqüenciamento de rRNA 16S como Pseudomonas putida, Pseudomonas sp e Pseudomonas fulva. As estirpes foram submetidas a testes bioquímicos de plantas e todos os atributos PGPR foram verificados na presença de pesticidas (clorpirifos e piriproxifeno). O índice de solubilização de fosfato da estirpe Ros2 foi mais elevado no meio NBRIP, isto é, 2,23 mm. Todas as estirpes apresentaram um pH ácido (varia de 2,5-5) em ambos os meios, isto é PVK e NBRIP. A estirpe Ros2 foi altamente positiva para a produção de amoníaco, bem como a produção de sideróforos enquanto a estirpe Rad2 foi positiva para a produção de HCN. Os resultados obtidos pelas estirpes Rad1, Rad2 e Ros2 para a produção de auxina foram 33,1, 30,67 e 15,38 μg ml-1 , respectivamente. A deformação Rad1 mostrou aumento de 16% na germinação percentual em comparação com o controlo na presença de stress de pesticida. Os resultados mais promissores para a estimativa do teor de clorofila foram obtidos na presença de carotenóides até 6 mgg-1 sem estresse por ambas as cepas Rad1 e Rad2. Estudo sugere que especialmente a estirpe Ros2 pode melhorar parâmetros de crescimento de plantas no estresse de pesticidas.

Knock-down the expression of TaH2B-7D using virus-induced gene silencing reduces wheat drought tolerance

BACKGROUND: Drought is a major abiotic stress affecting global wheat (Triticum aestivum L.) production. Exploration of drought-tolerant genes is essential for the genetic improvement of drought tolerance in wheat. Previous studies have shown that some histone encoding genes are involved in plant drought tolerance. However, whether the H2B family genes are involved in drought stress response remains unclear. METHODS: Here, we identified a wheat histone H2B family gene, TaH2B-7D, which was significantly up-regulated under drought stress conditions. Virus-induced gene silencing (VIGS) technology was used to further verify the function of TaH2B-7D in wheat drought tolerance. The phenotypic and physiological changes were examined in the TaH2B-7D knock-down plants. RESULTS: In the TaH2B-7D knock-down plants, relative electrolyte leakage rate and malonaldehyde (MDA) content significantly increased, while relative water content (RWC) and proline content significantly decreased compared with those in the non-knocked-down plants under drought stress conditions. TaH2B-7D knock-down plants exhibited severe sagging, wilting and dwarf phenotypes under drought stress conditions, but not in the non-knocked-down plants, suggesting that the former were more sensitive to drought stress. CONCLUSION: These results indicate that TaH2B-7D potentially plays a vital role in conferring drought tolerance in wheat.

PDIL1-2 can indirectly and negatively regulate expression of the AGPL1 gene in bread wheat

BACKGROUND: ADP-glucose pyrophosphorylase (AGPase), the key enzyme in plant starch biosynthesis, is a heterotetramer composed of two identical large subunits and two identical small subunits. AGPase has plastidial and cytosolic isoforms in higher plants, whereas it is mainly detected in the cytosol of grain endosperms in cereal crops. Our previous results have shown that the expression of the TaAGPL1 gene, encoding the cytosolic large subunit of wheat AGPase, temporally coincides with the rate of starch accumulation and that its overexpression dramatically increases wheat AGPase activity and the rate of starch accumulation, suggesting an important role. METHODS: In this study, we performed yeast one-hybrid screening using the promoter of the TaAGPL1 gene as bait and a wheat grain cDNA library as prey to screen out the upstream regulators of TaAGPL1 gene. And the barley stripe mosaic virus-induced gene-silencing (BSMV-VIGS) method was used to verify the functional characterization of the identified regulators in starch biosynthesis. RESULTS: Disulfide isomerase 1-2 protein (TaPDIL1-2) was screened out, and its binding to the TaAGPL1-1D promoter was further verified using another yeast one-hybrid screen. Transiently silenced wheat plants of the TaPDIL1-2 gene were obtained by using BSMV-VIGS method under field conditions. In grains of BSMV-VIGS-TaPDIL1-2-silenced wheat plants, the TaAGPL1 gene transcription levels, grain starch contents, and 1000-kernel weight also significantly increased. CONCLUSIONS: As important chaperones involved in oxidative protein folding, PDIL proteins have been reported to form hetero-dimers with some transcription factors, and thus, our results suggested that TaPDIL1-2 protein could indirectly and negatively regulate the expression of the TaAGPL1 gene and function in starch biosynthesis.

Combined application of bio-organic phosphate and phosphorus solubilizing bacteria (Bacillus strain MWT 14) improve the performance of bread wheat with low fertilizer input under an arid climate

Abstract This study was aimed to investigate the effect of bio-organic phosphate either alone or in combination with phosphorus solubilizing bacteria strain (Bacillus MWT-14) on the growth and productivity of two wheat cultivars (Galaxy-2013 and Punjab-2011) along with recommended (150-100 NP kg ha−1) and half dose (75-50 NP kg ha−1) of fertilizers. The combined application of bio-organic phosphate and the phosphorous solubilizing bacteria strain at either fertilizer level significantly improved the growth, yield parameters and productivity of both wheat cultivars compared to non-inoculated control treatments. The cultivar Punjab-2011 produced the higher chlorophyll contents, crop growth rate, and the straw yield at half dose of NP fertilizer; while Galaxy-2013, with the combined application of bio-organic phosphate and phosphorous solubilizing bacteria under recommended NP fertilizer dose. Combined over both NP fertilizer levels, the combined use of bio-organic phosphate and phosphorous solubilizing bacteria enhanced the grain yield of cultivar Galaxy-2013 by 54.3% and that of cultivar Punjab-2011 by 83.3%. The combined application of bio-organic phosphate and phosphorous solubilizing bacteria also increased the population of phosphorous solubilizing bacteria, the soil organic matter and phosphorous contents in the soil. In conclusion, the combined application of bio-organic phosphate and phosphorous solubilizing bacteria offers an eco-friendly option to harvest the better wheat yield with low fertilizer input under arid climate.

Cost-effective production of bacterial cellulose using acidic food industry by-products

Abstract To reduce the cost of obtaining bacterial cellulose, acidic by-products of the alcohol and dairy industries were used without any pretreatment or addition of other nitrogen sources. Studies have shown that the greatest accumulation of bacterial cellulose (6.19 g/L) occurs on wheat thin stillage for 3 days of cultivation under dynamic conditions, which is almost 3 times higher than on standard Hestrin and Schramm medium (2.14 g/L). The use of whey as a nutrient medium makes it possible to obtain 5.45 g/L bacterial cellulose under similar conditions of cultivation. It is established that the pH of the medium during the growth of Gluconacetobacter sucrofermentans B-11267 depends on the feedstock used and its initial value. By culturing the bacterium on thin stillage and whey, there is a decrease in the acidity of the waste. It is shown that the infrared spectra of bacterial cellulose obtained in a variety of environments have a similar character, but we found differences in the micromorphology and crystallinity of the resulting biopolymer.

Screening and characterization of endophytic Bacillus and Paenibacillus strains from medicinal plant Lonicera japonica for use as potential plant growth promoters

Abstract A total of 48 endophytic bacteria were isolated from surface-sterilized tissues of the medicinal plant Lonicera japonica, which is grown in eastern China; six strains were selected for further study based on their potential ability to promote plant growth in vitro (siderophore and indoleacetic acid production). The bacteria were characterized by phylogenetically analyzing their 16S rRNA gene similarity, by examining their effect on the mycelial development of pathogenic fungi, by testing their potential plant growth-promoting characteristics, and by measuring wheat growth parameters after inoculation. Results showed that the number of endophytic bacteria in L. japonica varied among different tissues, but it remained relatively stable in the same tissues from four different plantation locations. Among the three endophytic strains, strains 122 and 124 both had high siderophore production, with the latter showing the highest phosphate solubilization activity (45.6 mg/L) and aminocyclopropane-1-carboxylic acid deaminase activity (47.3 nmol/mg/h). Strain 170 had the highest indoleacetic acid (IAA) production (49.2 mg/L) and cellulase and pectinase activities. After inoculation, most of the six selected isolates showed a strong capacity to promote wheat growth. Compared with the controls, the increase in the shoot length, root length, fresh weight, dry weight, and chlorophyll content was most remarkable in wheat seedlings inoculated with strain 130. The positive correlation between enzyme (cellulose and pectinase) activity and inhibition rate on Fusarium oxysporum, the IAA production, and the root length of wheat seedlings inoculated with each tested endophytic strain was significant in regression analysis. Deformity of pathogenic fungal mycelia was observed under a microscope after the interaction with the endophytic isolates. Such deformity may be directly related to the production of hydrolytic bacterial enzymes (cellulose and pectinase). The six endophytic bacterial strains were identified to be Paenibacillus and Bacillus strains based on the results of 16S rRNA gene sequencing analysis and their physiological and biochemical characteristics. Results indicate the promising application of endophytic bacteria to the biological control of pathogenic fungi and the improvement of wheat crop growth.

Poly R decolorization and APPL production by Streptomyces violaceoruber and Streptomyces spiroverticillatus

Two mesophilic streptomycetes (S. violaceoruber and S. spiroverticillatus) were selected to study their Poly R-478 decolorization ability and lignocellulose solubilizing activity. Both strains were able to degrade Poly R-478 dye and ferulic acid during growth on a minimal salts medium. The Poly R-478 decolorizing activities of both strains were induced by adding different carbon sources to the culture media. S. violaceoruber could decolorize 63% of Poly R-478 after 24 h. Both strains could solubilize straw and produce acid-precipitable polymeric lignin (APPL) with different efficiency. From the major extracellular enzymes recovery of both strains on rice and wheat straw, we can predicate that the biodegradation process was partial indicating a possible utilization in biological delignification.

Manganese influx and its utilization efficiency in wheat.

Manganese deficiency in wheat has become an important nutritional disorder particularly in alkaline calcareous soils where rice-wheat rotation is followed. This experiment was aimed to study the mechanism of Mn efficiency during various developmental stages in six wheat cultivars grown at two Mn levels viz. 0 and 50 mg Mn kg-1soil (Mnapplied as MnSO4.H20) in pots. The Mn vegetative efficiency calculated on the basis of shoot dry weight at anthesis indicated HD 2967 and PBW 550 (bread wheat) as Mn efficient and durums as Mn inefficient. The efficient cultivars recorded highest values for influx, uptake, shoot dry weight, leaf area/plant, SPAD index, Fv/Fmratio and root length that explained their higher efficiencies whereas inefficiency of durum cultivars was attributed to their smaller roots and lower influx. Under Mn deficiency, PDW 314 and PDW 291 retained 68% and 64%, respectively, of total Mn uptake in vegetative parts (stem and leaves) and lowest in grains 7% and 5%, respectively, whereas PBW 550, BW 9178 and HD 2967 retained 29, 37 and 34% in vegetative parts, and 21, 17 and 15 % in grains, respectively at maturity. Higher utilization efficiency of efficient genotypes also indicated that increased Mn uptake with Mn supply produced more efficiently grains in efficient genotypes but vegetative parts in inefficient genotypes. Hence Mn efficiency of a cultivar could be explained by longer roots, higher uptake, influx and efficiency index during vegetative phase and higher grain yield and utilization efficiency during generative phase.

Ethanol production from agricultural wastes using Sacchromyces cervisae

The main objective of this study was production of ethanol from three lignocellulosic biomasses like sugarcane bagasse, rice straw and wheat straw by Sacchromyces cervisae. All the three substrates were ground to powder form (2 mm) and pretreated with 3%H2O2 + 2% NaOH followed by steaming at 130 °C for 60 min. These substrates were hydrolyzed by commercial cellulase enzyme. The whole fermentation process was carried out in 500 mL Erlenmeyer flask under anaerobic conditions in submerged fermentation at 30 °C for three days of incubation period. FTIR analysis of the substrates indicated significant changes in the alteration of the structure occurred after pretreatment which leads to efficient saccharification. After pretreatment the substrates were hydrolyzed by commercial cellulase enzyme and maximum hydrolysis was observed in sugarcane bagasse (64%) followed by rice straw (40%) and wheat straw (34%). Among all these tested substrates, sugarcane bagasse (77 g/L) produced more ethanol as compared to rice straw (62 g/L) and wheat straw (44 g/L) using medium composition of (%) 0.25 (NH4)2SO4, 0.1 KH2PO4, 0.05 MgSO4, 0.25 Yeast extract by S. cervisae.

Antioxidant response and Lea genes expression under salt stress and combined salt plus water stress in two wheat cultivars contrasting in drought tolerance.

Two wheat cultivars, C306 and PBW343 contrasting in drought tolerance were compared for their antioxidant response and Lea genes’ expression under salt stress (SS) and combined stress (CS) of salt stress plus water stress during seedlings growth. The drought susceptible cultivar (PBW343) behaved different towards SS/CS than towards WS. It accumulated more dry masses in shoots, more ascorbate, had higher ascorbate to dehydroascorbate ratio, lesser dehydroascorbate, lesser malondialdehyde (MDA), more proline and higher antioxidant enzymes under SS than under WS. CS increased dry masses, ascorbate, ascorbate to dehydroascorbate ratio, antioxidant enzymes and decreased dehydroascorbate and MDA contents from levels under WS. The drought tolerant cultivar (C306) though showed higher levels of ascorbate, ascorbate to dehydroascorbate ratio, lower levels of dehydroascorbate, showed lesser dry biomasses in shoots, higher MDA and lesser ascorbate peroxidase and catalase activities under SS than under WS and these features were improved on combining WS with SS. All lea genes were induced under all stresses in both cultivars except Wrab17 in C306 only, was not induced under any stress. Eight Lea genes out of ten were induced higher under WS than SS in C306 but induced same in PBW343. Wdhn13 gene was higher salt-responsive than other lea genes in both cultivars.

Individual and interactive effects of elevated carbon dioxide and ozone on tropical wheat (Triticum aestivum L.) cultivars with special emphasis on ROS generation and activation of antioxidant defence system.

The effects of elevated CO2 and O3, singly and in combination were investigated on various physiological, biochemical and yield parameters of two locally grown wheat (Triticum aestivum L.) cultivars (HUW-37 and K-9107) in open top chambers (OTCs). Elevated CO2 stimulated photosynthetic rate (Ps) and Fv/Fm ratio and reduced the stomatal conductance (gs). Reactive oxygen species, lipid peroxidation, anti-oxidative enzymes, ascorbic acid and total phenolics were higher, whereas Ps, gs, Fv/Fm, protein and photosynthetic pigments were reduced in elevated O3 exposure, as compared to their controls. Under elevated CO2 + O3, elevated levels of CO2 modified the plant performance against O3 in both the cultivars. Elevated CO2 caused significant increase in economic yield. Exposure to elevated O3 caused significant reduction in yield and the effect was cultivar-specific. The study concluded that elevated CO2 ameliorated the negative impact of elevated O3 and cultivar HUW-37 was more sensitive to elevated O3 than K-9107.

Characterization of differentially expressed stress-associated proteins in starch granule development under heat stress in wheat (Triticum aestivum L.).

Abiotic stress causes abrupt increase in the expression of stress-associated proteins, which provide tolerance by modulating the defense mechanism of plants. Small heat shock proteins (sHSPs) and anti-oxidant enzymes are important for environmental stress tolerance of the plants. In this study, two full-length cDNAs encoding small heat shock protein (sHSP) and superoxide dismutase (SOD), designated as TasHSP and SODI were identified and characterized from C-306 (thermotolerant) and PBW343 (thermosusceptible) cultivars of wheat (Triticum aestivum L.). An alpha crystalline domain was observed in TasHSP and manganese/iron binding domain in case of SODI. Quantitative real-time PCR showed very high transcript level of TasHSP and SOD in C-306 compared to PBW343 at different stages of growth and against differential heat stress (HS). Under differential HS at milky-dough stage, the fold change in transcript of both TasHSP and SOD was observed maximum in C-306, compared to PBW343. Protein profiling and isoenzymes analysis showed the expression of several heat-stable proteins and prominent isoenzymes of SOD in C-306, compared to PBW343. Scanning electron microscopy (SEM) of starch granules showed globular, well-shaped and more numbers of endospermic cells in C-306, compared to defragmented, irregular shaped and shrunken granules in case of PBW343 under HS treatment (42°C for 2 h). Diurnal change in soluble starch synthase (SSS) activity showed an increase in the activity during afternoon (35°C), compared to morning (29°C) and evening (32°C) in both the cultivars. Under heat stress (42°C for 2 h), a drastic decrease in the SSS activity was observed, due to the thermal denaturation of the enzyme. Thermotolerance capacity analyzed using cell membrane stability (CMS) showed significantly higher CMS in case of C-306, compared to PBW343 at different stages of growth. Findings suggest that abundance of TasHSP and SODI during milky-dough stage plays a very important role in starch granule biosynthesis. The mechanism may be further exploited to develop tolerant wheat cultivar with high quality seeds.

High density process to cultivate Lactobacillus plantarum biomass using wheat stillage and sugar beet molasses

Background: Owing to the growing interest in biofuels, the concept of a biorefinery where biomass is converted to a variety of useful products is gaining ground. We here present how distillery waste is combined with a by-product from a sugar production, molasses, to form a medium for the growth of Lactobacillus plantarum with yields and biomass densities comparable with conventional industrial media. Such approach enables a cost-effective utilization of the problematic wastewater from ethanol and a by-product from sugar production. It is the first approach that attempts to find low-cost media for the production of Lactobacillus plantarum biomass. Results: This study suggests that sieved wheat stillage enriched by adding 1.77 g/l yeast extract and 10 percent molasses (v/v), with NH4OH used for pH adjustment, may be used as a media for large-scale cultivation of L. plantarum. Such composition of the medium permits a high density of lactic acid bacteria (1.6 x 10(10) cfu/ml) to be achieved. Conclusions: The use of a fermentation medium consisting of distillery wastewater and molasses to obtain value-added products (such as LAB biomass and lactic acid) is a possible step for classical ethanol production to move towards a biorefinery model production in which all by and waste products are utilized to increase produced values and reduce waste production. This enables a cost-effective utilization of the problematic wastewater from ethanol and sugar production.

Continuous production of polygalacturonases [PGases] by Aspergillus awamori using wheat flour in surface culture fermentation

The continuous production of polygalacturonases [PGases] that Exo-polygalacturonase [exo-PGase] and Endo-polygalacturonase [endo-PGase] are two members of this group by a strain of Aspergillus awamori in surface culture fermentation was investigated. Surface culture fermentation is usually done in batch mode. Wheat flour acted as a good substrate for the cultivation of the fungus and production of PGases in surface culture fermentation. Fermentation started in batch mode until mycelia completely occupied the medium following growth of the microorganism, after which it was turned to the continuous mode by the introduction of fresh feed. The process continued for 34 days, and the thickness of the microbial layer on the surface of the liquid medium became almost constant after approximately one week. The production of PGases, however, continued throughout the experiment, and maximum activities of 1.2 U/ml and 0.014 U/ml were obtained for exo-polygalacturonase [exo- PGase] and endo-polygalacturonase [endo-PGase], respectively. An increase in production was observed when a similar system was used with a line for medium recycling. Lowering the residence time to 12 h decreased the exo-PGase and endo-PGase activities. Reducing the residence time from 24 h to 12 h almost halved the concentrations of the enzymes at the outlet

Characterization of cellulolytic activities of Bjerkandera adusta and Pycnoporus sanguineus on solid wheat straw medium / Caracterización de las actividades celulolíticas de Bjerkandera adusta y Pycnoporus sanguineus en un medio sólido de paja de trigo

Cellulolytic properties of two white rot fungi, Bjerkandera adusta and Pycnoporus sanguineus, cultivated on wheat straw agar medium, were characterized and compared. Optimal growing parameters for maximum enzyme production for both fungi were wheat straw medium pH 5 and 28ºC. B. adusta showed, on the 6th day of culture, carboxymethylcellulose (CMC)ase activity levels 1.6 times higher than maximal P. sanguineus activity, achieved on the 8th day. B. adusta supernatants also displayed higher activity levels towards xylan (3.6-fold) compared to those of P. sanguineus. However, enzymes from P. sanguineus were more robust resisting one hour incubation at high temperatures (up to 80ºC), and exhibiting activity and stability in pH range from 2 to 8. Cellulolytic activities, with molecular masses ranging from 25 to 90 kDa, from the two species were detected in zymograms.

Effect of mercury ion on the stability of the lipid-protein complex of isolated chloroplasts.

Mercury is known to interact with different parts of living systems causing serious biochemical and physiological disorder. In order to know the effect of mercury (Hg2+) ion on chloroplasts, the cell free organelle are incubated in an isotonic buffer medium in presence of mercury ion. The metal ion is found to induce membrane lipid peroxidation, loss of photosynthetic pigments and degradation of proteins. Such degradation brings about a drastic modification of lipid-protein organization of chloroplasts as reflected from a blue shift of absorption peaks and lowering of chlorophyll-a fluorescence intensity. The detrimental effect of Hg2+ ion has been explained in terms of direct binding with lipid-protein complex of photosynthetic membrane. Such a binding of metal ion exposes the lipid-protein complex for an easier entry and attack of reactive oxygen species (ROS) generated during incubation of chloroplasts in light and dark, thereby resulting in higher disorganization, which is evident from cation- induced changes in absorption and emission characteristics of the organelle.

Establishment of an efficient callus induction and plant regeneration system in Pakistani wheat (Triticum aestivum) cultivars

Four commercially grown wheat varieties of Pakistan, namely Inqilab-91, Chakwal-97, Tatara and Manthar were used for this investigation. For callus induction different concentrations of 2,4-Dichlorophenoxyaceticacid (2,4-D) along with 0.1 mg/L of Kinetin were evaluated. For regeneration initially different concentrations of Indole-3-Acetic Acid (IAA) and 6-BenzylAminoPurine (BAP) were tested. Best hormone combinations were further subjected to Kinetin and 6-ã-ã-dimethylallylaminopurine (2iP). For Inqilab-91, Chakwal-97 and Manthar, 3 mg/L of 2,4-D was found optimum, which induced 83.25 percent, 77.75 percent and 95.20 percent of embryogenic calli, respectively. Maximum callus induction (97.18 percent) was observed in Tatara when 2 mg/L of 2,4-D was used. As regard to regeneration, Inqilab-91, Chakwal-97 and Manthar showed maximum regeneration on media containing 0.1 mg/L IAA, 0.4 mg/L Kinetin and 0.5 mg/L 2iP, regenerating 87.25 percent, 81.75 percent and 68.75 percent respectively. For Tatara maximum regeneration of 12.25 percent was obtained on 0.1 mg/L IAA and 2 mg/L of BAP. Presently optimized regeneration method holds promise for facilitating the deployment of agronomical important trait through genetic transformation for the improvement of this important food crop.