Simultaneous production of intracellular triacylglycerols and extracellular polyol esters of fatty acids by <Emphasis Type="Italic">Rhodotorula babjevae</Emphasis> and <Emphasis Type="Italic">Rhodotorula</Emphasis> aff. <Emphasis Type="Italic">paludigena</Emphasis>

Microbial oils have been analyzed as alternatives to petroleum. However, just a handful of microbes have been successfully adapted to produce chemicals that can compete with their petroleum counterparts. One of the reasons behind the low success rate is the overall economic inefficiency of valorizing a single product. This study presents a lab-scale analysis of two yeast species that simultaneously produce multiple high-value bioproducts: intracellular triacylglycerols (TG) and extracellular polyol esters of fatty acids (PEFA), two lipid classes with immediate applications in the biofuels and surfactant industries. At harvest, the yeast strain Rhodotorula aff. paludigena UCDFST 81-84 secreted 20.9 ± 0.2 g L^?1 PEFA and produced 8.8 ± 1.0 g L^?1 TG, while the yeast strain Rhodotorula babjevae UCDFST 04-877 secreted 11.2 ± 1.6 g L^?1 PEFA and 18.5 ± 1.7 g L^?1 TG. The overall glucose conversion was 0.24 and 0.22 g_{(total lipid)} g _(glucose) ^?1 , respectively. The results present a stable and scalable microbial growth platform yielding multiple co-products.     

Isolation of Phosphate Solubilizing Bacteria from Maize Rhizosphere and Their Potential for Rock Phosphate Solubilization–Mineralization and Plant Growth Promotion

Phosphate solubilizing bacteria (PSB) play a significant role in plant P nutrition by their effect on soil P dynamics and their subsequent ability to make P available to plants via solubilization and mineralization processes. This study aimed to evaluate the effect of separate and combined use of indigenous PSB, poultry manure (PM) and compost on solubilization and mineralization of rock phosphate (RP) and their subsequent effect on growth and P accumulation of maize (Zea mays L.). A group of fifty seven bacteria were isolated from the rhizosphere/rhizoplane of maize that had been grown in soils collected from varying altitudes (655–2,576 m) of the mountain region of Rawalakot, Azad Jammu and Kashmir, Pakistan. After screening, the capacity of eleven isolates to solubilize mineral phosphate was quantitatively evaluated using insoluble Ca₃(PO₄)₂ in culture medium as a time course study through spectrometer. The growth hormone producing (IAA) capacity of the isolates was also determined. Furthermore, five potential isolates were tested for their ability to increase P release capacity (mineralization) of insoluble RP in an incubation study. The effect of PSB inoculation on maize was determined in a completely randomized greenhouse experiment where root and shoot biomass and P accumulation in plants were assessed. The P solubilization index of selected isolates varied from 1.94 to 3.69, while the P solubilization efficiency ranged between 94.1% and 269.0%. The isolates MRS18 and MRS27 displayed the highest values. The P solubilization in the liquid medium was maximum at 6 and 9 days of incubation ranging between 9.91 and 44.04 µgmL^?1 and the isolates MRS27 and MRS34 exhibited the highest solubilization. Six isolates showed additional capability of producing IAA ranging between 2.66 and 28.41 µgmL^?1. Results of the incubation study indicated that P release capacity (P mineralization) of RP-amended soil varied between 6.0 and 11.8 µgPg^?1 that had been significantly increased to 30.6–36.3 µgPg^?1 (maximum value) when PSB were combined with RP. The combined application of PSB and organic amendments (PM, compost) with RP further increased P mineralization by releasing a maximum of 37.7 µgPg^?1 compared with separate application of RP (11.8 µgPg^?1) and organic amendments (21.5 and 16.5 µgPg^?1). The overall effect of PSB (as a group) with RP over RP alone on maize growth showing a relative increase in shoot length 21%, shoot fresh weight 42%, shoot dry weight 24%, root length 11%, root fresh weight 59%, root dry weight 35% and chlorophyll content 32%. This study clearly indicates that use of PSB, and organic amendments with insoluble RP could be a promising management strategy to enhance P availability in soil pool and improve plant growth in intensive cropping systems.     

Stabilization of dextransucrase from <Emphasis Type="Italic">Leuconostoc mesenteroides</Emphasis> NRRL B-640

Phosphate solubilizing yeast (PSY) were isolated from rhizosphere, non-rhizosphere and fruits from Bhavnagar district. The potential of 25 yeasts were analyzed on the basis of phosphate solubilizing zone to growth on solid medium denoted as solubilization index (SI) which ranged from 1.10 to 1.50. Among 25 yeast isolates, 6 yeast belonging to genus Saccharomyces (2), Hansenula, Klockera, Rhodotorula and Debaryomyces exhibited highest SI (1.33–1.50) were further examined for in vitro tricalcium phosphate (TCP) and low grade rock phosphate (RP) solubilization. TCP proved superior to RP with all the yeasts. Within low grade RPs tested, except isolate Y5, all isolates showed maximum solubilization with Hirapur RP (HRP) ranging from 7.24 to 19.30 mg% P₂O₅. Among six PSY screened, Debaryomyces hansenii showing maximal HRP solubilization was chosen for further physiological studies. Maximum HRP solubilization was expressed in following condition: pH optima 7.0, temperature optima 28°C and optimal period of incubation were 15 days. Acidic pH of the spent media was a constant feature in all the cases. No correlation could be established between final acidity produced by yeasts and the quantity of phosphate liberated.     

Phosphate solubilization potential and modeling of stress tolerance of rhizobacteria from rice paddy soil in northern Iran

The purposes of this study were to evaluate the phosphate solubilization activity of bacteria isolated from the rhizosphere of rice paddy soil in northern Iran, and to study the effect of temperature, NaCl and pH on the growth of these isolates by modeling. Three of the most effective strains from a total of 300 isolates were identified and a phylogenetic analysis was carried out by 16S rDNA sequencing. The isolates were identified as Pantoea ananatis (M36), Rahnella aquatilis (M100) and Enterobacter sp. (M183). These isolates showed multiple plant growth-promoting attributes such as phosphate solubilization activity and indole-3-acetic acid (IAA) production. The M36, M100 and M183 isolates were able to solubilize 172, 263 and 254 µg ml^?1 of Ca₃(PO₄)₂ after 5 days of growth at 28 °C and pH 7.5, and to produce 8.0, 2.0 and 3.0 μg ml^?1 of IAA when supplemented with l-tryptophan (1 mg ml^?1) for 72 h, at 28 °C and pH 7.0, respectively. The solubilization of insoluble phosphate was associated with a drop in the pH of the culture medium and there was an inverse relationship between pH and solubilized P (r = ?0.98, P < 0.0952). There were no significant differences among isolates in terms of acidity tolerance based on their confidence limits as assessed by segmented model analysis and all isolates were able to grow at pH 4.3–11 (with optimum at 7.0–7.5). Based on a sigmoidal trend of a three-parameter logistic model, the salt concentration required for 50 % inhibition was 8.15, 6.30 and 8.23 % NaCl for M36, M100 and M183 isolates, respectively. Moreover, the minimum and maximum growth temperatures estimated by the segmented model were 5.0 and 42.75 °C for M36, 12.76 and 40.32 °C for M100, and 10.63 and 43.66 °C for M183. The three selected isolates could be deployed as inoculants to promote plant growth in an agricultural environment.     

Fungal rock phosphate solubilization using sugarcane bagasse

The effects of different doses of rock phosphate (RP), sucrose, and (NH₄)₂SO₄ on the solubilization of RP from Araxá and Catal?o (Brazil) by Aspergillus niger, Penicillium canescens, Eupenicillium ludwigii, and Penicillium islandicum were evaluated in a solid-state fermentation (SSF) system with sugarcane bagasse. The factors evaluated were combined following a 2³?+?1 factorial design to determine their optimum concentrations. The fitted response surfaces showed that higher doses of RP promoted higher phosphorus (P) solubilization. The addition of sucrose did not have effects on P solubilization in most treatments due to the presence of soluble sugars in the bagasse. Except for A. niger, all the fungi required high (NH₄)₂SO₄ doses to achieve the highest level of P solubilization. Inversely, addition of (NH₄)₂SO₄ was inhibitory to P solubilization by A. niger. Among the fungi tested, A. niger stood out, showing the highest solubilization capacity and for not requiring sucrose or (NH₄)₂SO₄ supplementation. An additional experiment with A. niger showed that the content of soluble P can be increased by adding higher RP doses in the medium. However, P yield decreases with increasing RP doses. In this experiment, the maximal P yield (approximately 60?%) was achieved with the lower RP dose (3?g?L^?1). Our results show that SSF can be used to obtain a low cost biofertilizer rich in P combining RP, sugarcane bagasse, and A. niger. Moreover, sugarcane bagasse is a suitable substrate for SSF aiming at RP solubilization, since this residue can supply the C and N necessary for the metabolism of A. niger within a range that favors RP solubilization.     

Biochar Enhances Aspergillus niger Rock Phosphate Solubilization by Increasing Organic Acid Production and Alleviating Fluoride Toxicity

During fungal rock phosphate (RP) solubilization, a significant quantity of fluoride (F⁻) is released together with phosphorus (P), strongly inhibiting the process. In the present study, the effect of two F⁻ adsorbents [activated alumina (Al₂O₃) and biochar] on RP solubilization by Aspergillus niger was examined. Al₂O₃ adsorbed part of the F⁻ released but also adsorbed soluble P, which makes it inappropriate for microbial RP solubilization systems. In contrast, biochar adsorbed only F⁻ while enhancing phosphate solubilization 3-fold, leading to the accumulation of up to 160 mg of P per liter. By comparing the values of F⁻ measured in solution at the end of incubation and those from a predictive model, it was estimated that up to 19 mg of F⁻ per liter can be removed from solution by biochar when added at 3 g liter⁻¹ to the culture medium. Thus, biochar acted as an F⁻ sink during RP solubilization and led to an F⁻ concentration in solution that was less inhibitory to the process. In the presence of biochar, A. niger produced larger amounts of citric, gluconic, and oxalic acids, whether RP was present or not. Our results show that biochar enhances RP solubilization through two interrelated processes: partial removal of the released F⁻ and increased organic acid production. Given the importance of organic acids for P solubilization and that most of the RPs contain high concentrations of F⁻, the proposed solubilization system offers an important technological improvement for the microbial production of soluble P fertilizers from RP.     

Expression of exoinulinase genes in Saccharomyces cerevisiae to improve ethanol production from inulin sources

To improve inulin utilization and ethanol fermentation, exoinulinase genes from the yeast Kluyveromyces marxianus and the recently identified yeast, Candida kutaonensis, were expressed in Saccharomyces cerevisiae. S. cerevisiae harboring the exoinulinase gene from C. kutaonensis gave higher ethanol yield and productivity from both inulin (0.38 vs. 0.34 g/g and 1.35 vs. 1.22 g l^?1 h^?1) and Jerusalem artichoke tuber flour (0.47 vs. 0.46 g/g and 1.62 vs. 1.54 g l^?1 h^?1) compared with the strain expressing the exoinulinase gene from K. marxianus. Thus, the exoinulinase gene from C. kutaonensis is advantageous for engineering S. cerevisiae to improve ethanol fermentation from inulin sources.     

Xylitol production by genetically modified industrial strain of <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis> using glycerol as co-substrate

Xylitol is commercially used in chewing gum and dental care products as a low calorie sweetener having medicinal properties. Industrial yeast strain of S. cerevisiae was genetically modified to overexpress an endogenous aldose reductase gene GRE3 and a xylose transporter gene SUT1 for the production of xylitol. The recombinant strain (XP-RTK) carried the expression cassettes of both the genes and the G418 resistance marker cassette KanMX integrated into the genome of S. cerevisiae. Short segments from the 5′ and 3′ delta regions of the Ty1 retrotransposons were used as homology regions for integration of the cassettes. Xylitol production by the industrial recombinant strain was evaluated using hemicellulosic hydrolysate of the corn cob with glucose as the cosubstrate. The recombinant strain XP-RTK showed significantly higher xylitol productivity (212 mg L^?1 h^?1) over the control strain XP (81 mg L^?1 h^?1). Glucose was successfully replaced by glycerol as a co-substrate for xylitol production by S. cerevisiae. Strain XP-RTK showed the highest xylitol productivity of 318.6 mg L^?1 h^?1 and titre of 47 g L^?1 of xylitol at 12 g L^?1 initial DCW using glycerol as cosubstrate. The amount of glycerol consumed per amount of xylitol produced (0.47 mol mol^?1) was significantly lower than glucose (23.7 mol mol^?1). Fermentation strategies such as cell recycle and use of the industrial nitrogen sources were demonstrated using hemicellulosic hydrolysate for xylitol production.     

Optimized culture medium for the production of flavonoids from <Emphasis Type="Italic">Orostachys cartilaginea</Emphasis> V.N. Boriss. callus cultures

The optimal culture medium for the production of flavonoid compounds from Orostachys cartilaginea V. N. Boriss. calluses was studied. In callus cultures of O. cartilaginea, the flavonoid monomer content, in decreasing order was kaempferol-3-O-rutinoside (Kp-3-rut), quercetin 3-O-glucoside (Qc-3-glc), epicatechin gallate (Ecg), kaempferide (Ke), and quercetin (Qc). The results of the uniform design experiment indicated that the production of Qc, Ke, Qc-3-glc, Kp-3-rut, and total flavonoids were satisfactory in callus grown on full salt strength (1×) of Murashige and Skoog (MS) medium supplemented with 3.5 mg L^?1 6-benzylaminopurine (BA) and 0.1 mg L^?1 1-naphthalene acetic acid (NAA). By contrast, only Ecg was found in callus grown on 0.75× MS medium supplemented with 1.5 mg L^?1 BA and 0.3 mg L^?1 NAA. A phosphate concentration of 1.25 mM in the MS medium favored the production of Qc and Ke, whereas 0.75 mM phosphate was optimal for the production of Ecg, Qc-3-glc, Kp-3-rup, and total flavonoids. The NH₄ ⁺/NO₃ ^? ratios of 30/30 mM in the MS medium promoted Ke, Ecg, Qc-3-glc, Kp-3-rup, and total flavonoid production. However, a NH₄ ⁺/NO₃ ^? ratio of 20/40 mM enhanced Qc production. The effect of sucrose concentrations on the accumulation of different flavonoid monomers was comparatively more regular. The flavonoid content increased as the sucrose concentration increased from 20 to 40 g L^?1, peaked at 40 g L^?1, and decreased at concentrations greater than 40 g L^?1. Therefore, 40 g L^?1 sucrose was optimal for the production of the five flavonoid monomers and total flavonoids. The present findings demonstrate the possibility of producing flavonoid compounds from O. cartilaginea callus.     

Culture Media Statistical Optimization for Biomass Production of a Ligninolytic Fungus for Future Rice Straw Degradation

The main objective of this study was to optimize a culture media for low scale biomass production of Pleurotus spp. Future applications of this optimization will be implemented for “in situ” rice straw degradation, increase soil nutrients availability, and lower residue and rice culture management costs. Soil samples were taken from different points in six important rice production cities in Colombia. For carbon and nitrogen source selection a factorial 4² design was carried out. The Plackett-Burman design permitted to detect carbon, nitrogen and inducer effects on fungus growth (response variable for all designs). This optimization was carried out by a Box-Behnken design. Finally a re-optimization assay for glucose concentration was performed by means of a One Factor design. Only 4/33 (12 %) isolates showed and important laccase or manganese peroxidase activity compared to Pleurotus ostreatus (HPB/P3). We obtained an increased biomass production in Pleurotus spp. (T1.1.) with glucose, followed by rice husk. Rice straw was considered an inducing agent for lignin degradation. Glucose was a significant component with positive effects, whereas Tween 80 and pH had negative effects. On the contrary, rice husk, yeast extract and CaCl₂ were not significant components for increase the biomass production. Final media composition consisted of glucose 25 g L^?1, yeast extract 5 g L^?1, Tween 80 0.38 % (v/v), Rice husk 10 g L^?1, CaCl₂ 1 g L^?1, and pH 4.88 ± 0.2. The Box-Behnken polynomial prediction resulted to be lower than the experimental validation of the model (6.59 vs. 6.91 Log₁₀ CFU ml^?1 respectively).     

Rock phosphate solubilization by psychrotolerant Pseudomonas spp. and their effect on lentil growth and nutrient uptake under polyhouse conditions

Psychrotolerant Pseudomonas isolates (RT5RP2 and RT6RP) isolated from the rhizoplane of wild grass at 3,100 and 3,800 m above mean sea level, respectively, from Rudraprayag district of Uttarakhand (India), were found to solubilize Udaipur rock phosphate (URP). Both isolates grew at temperatures ranging from 4 to 30 °C. Kinetics of phosphate solubilization by the bacterial strains showed a nonlinear regression of the rate of P solubilization, which fitted best in the power model, and showed a declining trend across three different temperatures. Under pot culture conditions, bacterization of lentil seeds (cv. VL Masoor 507) with the psychrotolerant Pseudomonas strains when combined with URP as a sole source of phosphorus resulting in significant enhancement in P uptake of the plants, compared to the application of rock phosphate alone.     

Study of the physicochemical parameters and spontaneous fermentation during the traditional production of yakupa,an indigenous beverage produced by Brazilian Amerindians

Yakupa is a traditional non-alcoholic cassava beverage produced by Brazilian Amerindians. In this work the microbial dynamics and metabolites involved in yakupa fermentation were investigated by PCR-denaturing gradient gel electrophoresis and chromatography analysis, respectively. The lactic acid bacteria (LAB) population was higher than yeast in the beginning of fermentation (5 log CFU mL^?1 and 3 log CFU mL^?1, respectively) and after 36 h both population increased reaching 7 log CFU mL^?1, remaining constant until 60 h. Culture dependent and independent methods in combination identified the bacteria Lactobacillus fermentum, L. plantarum, Weissela cibaria and W. confusa, and yeasts Saccharomyces cerevisiae and Pichia kudriavzevii. Maltose (41.2 g L^?1), ethanol (6.5 g L^?1) and lactic acid (7.8 g L^?1) were the most abundant compounds identified by high performance liquid chromatography. Aldehydes, acids, alcohols and esters were identified by gas chromatography flame ionization detection. By the metabolites and PCA analysis we may assign the beverage’s flavor to the microbial metabolism. Heterolactic LAB and S. cerevisiae dominated the yakupa fermentation, being responsible for the organoleptic characteristics of the final product. This is the first time that the microbial dynamics and physicochemical parameters were investigated in the yakupa beverage and it may contribute to the future selection of starter cultures to perform yakupa fermentations.     

Isolation,Selection and Characterization of Root-Associated Rock Phosphate Solubilizing Bacteria in Moroccan Wheat (Triticum aestivum L.)

Abstract

Phosphorus (P) is the most important macronutrient next to nitrogen for the growth and development of plants. But often unavailable for plants because of its high reactivity with many soil constituents. Thus, the use of phosphate solubilizing bacteria (PSBs) as biofertilizers seems to be an effective way to resolve the soluble phosphorus availability in soil. The present study was conducted to isolate and characterize rock PSB associated with the rhizosphere of wheat (Triticum aestivum L.) from fourteen different wheat-growing sites of Meknes region in Morocco. A total of one hundred ninety-eight (198) rock PSBs were isolated employing NBRIP medium amended with rock phosphate (RP), out of which five strains (A17, A81, B26, B106, and B107) were selected for their strong ability to dissolve RP and were tested in vitro for plant growth-promoting (PGP) traits including production of indole acetic acid (IAA), siderophores, hydrogen cyanide (HCN), and antifungal activity, as well as their response to the effect of extrinsic and intrinsic stress. The 16S rRNA gene sequencing and phylogenetic analysis identified these isolates belong to four genera, Pantoea, Pseudomonas, Serratia, and Enterobacter. The phosphate solubilization index (SI) of selected isolates ranged between 2.3 and 2.7, and the amount of solubilized phosphorus in the liquid medium varied from 59.1 to 90.2 µg mL^?1. HPLC analysis revealed that all the selected isolates produced multiple organic acids (oxalic, citric, gluconic succinic, and fumaric acids) from glucose under aerobic conditions. Except for the A81 strain, all selected isolates were able to produce IAA ranging between 2.9 and 21.2 µg mL^?1. The isolates A17, B26, and B107 showed the ability to produce siderophores ranging from 79.3 to 20.8% siderophore units. Only two strains (A17 and B26) were able to produce HCN. All selected isolates showed good resistance against different environmental stresses like 10–50?°C temperature, 0.5–2?g L^?1 salt concentration and 4.5–9?pH range, and against different antibiotics. The antagonistic effect showed that among the five selected strains, only two strains (B26 and A17) were able to suppress the growth of tested fungi. This study clearly indicates that our selected rock PSBs can be used as biofertilizers for grain crops after studying their interaction with the host crop and field evaluation.     

Cloning, expression and characterization of phenylalanine ammonia-lyase from Rhodotorula glutinis

The industrial-scale production of phenylalanine ammonia-lyase (PAL) mainly uses strains of Rhodotorula. However, the PAL gene from Rhodotorula has not been cloned. Here, the full-length gene of PAL from Rhodotorula glutinis was isolated. It was 2,121 bp, encoding a polypeptide with 706 amino acids and a calculated MW of 75.5 kDa. Though R. glutinis is an anamorph of Rhodosporium toruloides, the amino acid sequences of PALs them are not the same (about 74 % identity). PAL was expressed in E. coli and characterized. Its specific activity was 4.2 U mg^?1 and the k _cat/K _m was 1.9 × 10⁴ mM^?1 s^?1, exhibiting the highest catalytic ability among the reported PALs. The genetic and biochemical information reported here should facilitate future application in industry.     

Marine Actinobacteria Showing Phosphate-Solubilizing Efficiency in Chorao Island, Goa, India

The occurrence and distribution of an actinobacteria group of bacteria capable of dissolving insoluble phosphates were investigated in this study in marine environments, especially in sediments of Chorao Island, Goa Province, India. A total of 200 bacterial isolates of actinobacteria was isolated. All isolates were screened for phosphate-solubilizing activity on Pikovskaya’s agar. Thirteen different isolates exhibiting maximum formation of halos (zone of solubilization) around the bacterial colonies were selected for quantitative estimations of P-solubilization. Quantitative estimations for P-solubilization were analyzed for up to 10 days at intervals of 24 h. Maximum solubilization from 89.3 ± 3.1 to 164.1 ± 4.1 μg ml^?1 was observed after 6 days of incubation in six of all isolates, while the isolate NII-1020 showed maximum P-solubilization. The increase in solubilization coincided with the drop in pH. Many of these species showed wide range of tolerance to temperature, pH, and salt concentrations. Further, 16S rRNA gene sequence analyses were carried to identify the bacterial groups which are actively solubilized phosphate in vitro. Gene sequencing results reveal that all isolates were clustered into six different actinobacterial genera: Streptomyces, Microbacterium, Angustibacter, Kocuria, Isoptericola, and Agromyces. The presence of phosphate-solubilizing microorganisms and their ability to solubilize phosphate were indicative of the important role played by bacteria in the biogeochemical cycle of phosphorus and the plant growth in coastal ecosystems.     

Decreased mineral availability enhances rock phosphate solubilization efficiency in <Emphasis Type="Italic">Aspergillus niger</Emphasis>

Microbial solubilization of rock phosphate (RP) is mainly achieved by the production of organic acids and medium acidification through H⁺ release. During RP solubilization, mineral nutrient availability is likely to be critical for determining how much carbon is channeled either for metabolite synthesis or for microbial growth, influencing organic acid release by microorganisms. Thus, the objective of this work was to study the relationships between the concentration of mineral nutrients in the growth medium and the efficiency of RP solubilization by Aspergillus niger FS1. For this, the fungus was grown in Czapek medium containing 0, 1, 2, 10, 50, and 100 % of its original concentration of mineral nutrients. Decreasing mineral availability in the growth medium led to decreases in fungal biomass and solubilized P, and increases in titratable acidity and solubilization efficiency as expressed by mg solubilized P per g fungal biomass (Y_P/B), indicating a shift in fungal metabolism from biomass production to organic acid release. The transfer of pre-grown biomass to media with or without added minerals confirmed that lower mineral availability increases Y_P/B and led to the solubilization of 76 % of P present in Patos RP. These observations open new perspectives on improving RP solubilization systems by manipulating mineral nutrient availability in the medium, with consequent gains in cost reduction.     

Swainsonine,a novel fungal metabolite: optimization of fermentative production and bioreactor operations using evolutionary programming

The optimization of bioreactor operations towards swainsonine production was performed using an artificial neural network coupled evolutionary program (EP)-based optimization algorithm fitted with experimental one-factor-at-a-time (OFAT) results. The effects of varying agitation (300–500 rpm) and aeration (0.5–2.0 vvm) rates for different incubation hours (72–108 h) were evaluated in bench top bioreactor. Prominent scale-up parameters, gassed power per unit volume (P _g/V _L, W/m³) and volumetric oxygen mass transfer coefficient (K _L a, s^?1) were correlated with optimized conditions. A maximum of 6.59 ± 0.10 μg/mL of swainsonine production was observed at 400 rpm-1.5 vvm at 84 h in OFAT experiments with corresponding P _g/V_L and K _L a values of 91.66 W/m³ and 341.48 × 10^?4 s^?1, respectively. The EP optimization algorithm predicted a maximum of 10.08 μg/mL of swainsonine at 325.47 rpm, 1.99 vvm and 80.75 h against the experimental production of 7.93 ± 0.52 μg/mL at constant K _L a (349.25 × 10^?4 s^?1) and significantly reduced P _g/V _L (33.33 W/m³) drawn by the impellers.     

Phosphate solubilization by stress-tolerant soil fungus <Emphasis Type="Italic">Talaromyces funiculosus</Emphasis> SLS8 isolated from the Neem rhizosphere

A promising biotechnological strategy in the management of phosphorus (P) fertilization is the use of phosphate-solubilizing fungi to solubilize rock phosphates and allow the recovery of unavailable P fixed to soil particles. Phosphate-solubilizing rhizosphere fungus, Talaromyces funiculosus SLS8, isolated from Neem (Azadirachta indica) on saline soil, was tolerant to environmental stressors, salinity and agricultural systemic fungicides. Phosphate solubilization under different nutritional conditions was investigated by culturing T. funiculosus SLS8 in Pikovskaya liquid medium containing different nitrogen sources (ammonium sulfate, casein, urea, potassium nitrate or sodium nitrate) and carbon sources (glucose, fructose, galactose or sucrose), NaCl, and three systemic fungicides. The highest concentration of solubilised phosphate (187 mg P L^?1) was achieved after 5 days of incubation in the medium with glucose and ammonium sulphate. The culture pH decreased from 6.5 to 4.2 and HPLC demonstrated organic acid production. Phosphate solubilized was highly negatively correlated with pH (r?=??0.96). Increasing salinity had no effect on phosphate solubilization. The maximum tolerance limits to systemic fungicides carbendazim, mancozeb, and hexaconazole were 12.5 μg mL^?1, 2,000 μg mL^?1 and 250 μl mL^?1 respectively. At these concentrations carbendazim, mancozeb and hexaconazole were found to decrease phosphate solubilization by 55 %, 37 %, and 30 %, respectively. Our results indicate that T. funiculosus SLS8 may be a potential candidate for the development of a biofertilizer for maintaining available phosphate levels in environmentally stressed soils such as saline agricultural soils impacted by systemic fungicide application or seed treatment.     

The citric acid production from raw glycerol by Yarrowia lipolytica yeast and its regulation

The optimal cultivation conditions ensuring the maximal rate of citric acid (CA) biosynthesis by glycerol-grown mutant Yarrowia lipolytica NG40/UV7 were found to be as follows: growth limitation by inorganic nutrients (nitrogen, phosphorus, or sulfur), 28 °C, pH 5.0, dissolved oxygen concentration (pO₂) of 50 % (of air saturation), and pulsed addition of glycerol from 20 to 80 g L^?1 depending on the rate of medium titration. Under optimal conditions of fed-batch cultivation, in the medium with pure glycerol, strain Y. lipolytica NG40/UV7 produced 115 g L^?1 of CA with the mass yield coefficient of 0.64 g g^?1 and isocitric acid (ICA) amounted to 4.6 g L^?1; in the medium with raw glycerol, CA production was 112 g L^?1 with the mass yield coefficient of 0.90 g g^?1 and ICA amounted to 5.3 g L^?1. Based on the activities of enzymes involved in the initial stages of raw glycerol assimilation, the tricarboxylic acid cycle and the glyoxylate cycle, the mechanism of increased CA yield from glycerol-containing substrates in Y. lipolytica yeast was explained.     

Selection of phosphate-solubilizing diazotrophic Herbaspirillum and Burkholderia strains and their effect on rice crop yield and nutrient uptake

Background and Aims

Plant growth-promoting bacteria, mainly diazotrophs and phosphate solubilizers, can reduce the use of chemical fertilizers for rice crops. Here, diazotrophic bacteria isolated from rice were screened for their ability to solubilize inorganic P (P_i) in vitro and in association with rice plants cultivated in pots.

Methods

Forty-nine isolates were tested for the ability to solubilize P_i on NBRIP and GL agar plate media and seven selected strains were further evaluated in NBRIP liquid medium. Three of these strains were inoculated in rice plants grown in soil pots containing ¹⁵N-labeled fertilizer and two sources of P: tricalcium phosphate (TCP) or simple superphosphate (SSP). The dry matter, yield, N, P, and the ¹⁵N content accumulated in plant tissues were measured at 135 days after planting.

Results

Seven strains belonging to the genera Herbaspirillum and Burkholderia formed a halo of solubilized P_i on agar plates. The Burkholderia strains showed peak soluble P (around 200 mg P L^?1) on the fifth day when grown in NBRIP liquid medium for 14 days. Inoculation of Herbaspirillum strains (H18, ZA15) and a Burkholderia vietaminensis strain (AR114) increased rice grain yield from 33 to 47 % with TCP and 18 to 44 % with TSS, respectively. The bacterial inoculation led to enhanced N-use efficiency of the ¹⁵N-labeled fertilizer.

Conclusion

These results suggest that the selection and use of P-solubilizing diazotrophic bacteria are a good strategy to promote P solubilization and/or N use efficiency in rice plants.