Organic Nitrogen Sources Research Articles

Clavulanic Acid Overproduction: A Review of Environmental Conditions, Metabolic Fluxes, and Strain Engineering in Streptomyces clavuligerus

Clavulanic acid is a potent β-lactamase inhibitor produced by Streptomyces clavuligerus, widely used in combination with β-lactam antibiotics to combat antimicrobial resistance. This systematic review analyzes the most successful methodologies for clavulanic acid overproduction, focusing on the highest yields reported in bench-scale and bioreactor-scale fermentations. Studies have demonstrated that glycerol is the preferred carbon source for clavulanic acid production over other sources like starch and dextrins. The optimization of feeding strategies, especially in fed-batch operations, has improved glycerol utilization and extended the clavulanic acid production phase. Organic nitrogen sources, particularly soybean protein isolates and amino acid supplements such as L-arginine, L-threonine, and L-glutamate, have been proven effective at increasing CA yields both in batch and fed-batch cultures, especially when balanced with appropriate carbon sources. Strain engineering approaches, including mutagenesis and targeted genetic modifications, have allowed for the obtainment of overproducer S. clavuligerus strains. Specifically, engineering efforts that overexpress key regulatory genes such as ccaR and claR, or that disrupt competing pathways, redirect the metabolic flux towards CA biosynthesis, leading to high clavulanic acid titers. The fed-batch operation at the bioreactor scale emerges as the most feasible alternative for prolonged clavulanic acid production with both wild-type and mutant strains, allowing for the attainment of high titers during cultivations.

Effect of Organic and Inorganic Sources of Nitrogen on Growth, Yield, Quality and Soil Properties of Wheat (Triticum aestivum L.)

Background: The concept of combined use of both organic and inorganic fertilizer should be followed to prevent severe health hazards and to protect the environment. The combined use of organic and inorganic fertilizers is an effective way to maintain nutrient supply, gives organic carbon to soil microbes and mobilizes soil-bound nutrients on decomposition through the release of organic acid. Combined use refers to combining all available nutrient sources, such as organic and inorganic sources with components in a prudent manner to create an environment that is both environmentally sound and economically ideal for farming. Methods: The experiment was conducted at the research farm of the School of Agriculture, Department of Agronomy, Lovely Professional University, Phagwara (Punjab) during rabi season in the year 2021-2022. The objective of the study was to evaluate the "Effect of organic and inorganic source of nitrogen on growth and yield of wheat (Triticum aestivum L.)". Data recorded on different aspects of crop, viz., growth, yield attributes, yield, quality as well as soil chemical properties were recorded and subjected to statistical analysis. Result: Among various treatments 25% RDN with vermicompost + 75% RDN from inorganic fertilizers showed a significant impact on the growth and yield parameters of wheat crop. Significantly higher growth parameters viz., plant height, number of tillers m-2, dry matter and relative growth rate were observed in T12 (25% RDN with vermicompost + 75% RDN from inorganic fertilizers) of crop. It was also showed higher significant impact on yield attributes i.e., number of effective tillers-2, grain yield, straw yield, harvest index, quality parameters as well as soil parameter like available nitrogen, phosphorus and potassium in T12 (25% RDN with vermicompost + 75% RDN from inorganic fertilizers).

Mycelial mass, microbial lipids and γ-linolenic acid (GLA) by Cunninghamella elegans cultivated on agro-industrial residues

In the current study, the Zygomycetes fungus Cunninghamella elegans NRRL Y-1392 was evaluated for its ability to grow in extracts derived from dried and ground agricultural residues, such as mushroom stalks and roots from hydroponically cultivated lettuces and produce poly-unsaturated fatty acids (PUFA) and γ-linolenic acid (GLA) rich lipids. Initially, the compositions of stalks and lettuce roots were analysed, and the fungus was batch-flask cultivated on six different commercial semi-defined substrates containing different sugars detected in stalks and roots to evaluate its catabolic ability. C. elegans was capable to assimilate all sugars, but at a lower rate in the case of arabinose. Subsequently, C. elegans was cultivated on tailor-made semi-defined commercial substrates, resembling hydrolysates containing carbohydrates found in mushroom stalks, under both nitrogen-excess and nitrogen-limited conditions, and resembling that of hydrolysates of roots, under nitrogen-excess conditions. Based on the results, under nitrogen-excess conditions, in the case of media resembling stalks hydrolysates, higher production values for biomass, PUFAs, and GLA were observed (20.3 g/L, 1906 mg/L, 668 mg/L), accompanied by high productivity values due to short cultivation periods, while under nitrogen limitation, high lipid accumulation (lipid in dry cell weight =48%, w/w) was presented, and lipids rich in oleic acid were produced. Finally, the fungus was cultivated on a medium derived from hot water-extraction applied to mushroom stalks, enriched with organic nitrogen sources. The fungus was successfully grown on the sugar-rich water-extract derived from mushroom stalks, resulting in dry biomass of 14.5 g/L, lipids of 1.8 g/L, with 15% (w/w) of GLA in cellular lipids.

Enhancement of ε-poly-L-lysine production by Streptomyces albulus FQF-24 with feeding strategies using cassava starch as carbon source.

ε-Poly-L-lysine (ε-PL) is a natural and wide-spectrum antimicrobial additive. In this study, the production of ε-PL by Streptomyces albulus FQF-24 using cassava starch (CS) as carbon source and the effects of different feeding methods were investigated in a fermenter. The initial shake flask experiments demonstrated the efficient production of ε-PL with CS, achieving the ε-PL production of 1.18g/L. Subsequent investigations in the fermenter identified that the ideal pH was 3.8 during the ε-PL synthesis phase. Under this condition, the production of ε-PL reached 1.35g/L. When the pH was maintained at 3.8, the investigation of improvement of feeding composition was carried out in a 5 L fermenter. The intermittent feeding containing CS, inorganic and organic nitrogen sources resulted in the maximum ε-PL production and dry cell weight (DCW) reaching 17.17g/L and 42.73g/L. Additionally, continuous feeding with the composition of CS, organic and inorganic nitrogen sources, and inorganic salts further increased ε-PL production and DCW to 27.56g/L and 38.5g/L. Summarily, the above results indicate that the fermentation using low-cost CS and continuous feeding strategy with whole medium composition can provide a beneficial reference for the efficient production of ε-PL.

Quantifying particulate organic matter: source composition and fluxes at the river-estuary interface

Particulate organic matter (POM) characteristics and variability have been widely studied along the land-ocean aquatic continuum, yet, gaps remain in quantifying its source composition, fluxes, and dynamics at the river-estuary interface. POM in rivers consists of a complex mixture of sources, derived both from locally produced (i.e. phytoplankton) and from adjacent ecosystems (e.g. terrestrial POM). Each source differ in its trophic and biogeochemical characteristics, hence impacting its integration into local food webs, its transfer to estuaries and sea, and its contribution to biogeochemical processes. In this study, we use a robust approach based on in situ POM to characterize river POM end-members, to quantify POM composition and dynamics, and to identify the related key drivers. This study was performed at the River-Estuary interface of one of the main rivers in Western Europe (the Loire River, France). For 3 years, we conducted bimonthly measurements of carbon and nitrogen isotopic (δ13C, δ15N) and elemental (C/N) ratios to quantify the contribution of two sources (phytoplankton and terrestrial POM) to the POM mixture and calculated annual fluxes of particulate organic carbon (POC) and nitrogen (PN) sources. Throughout the year, POM consisted of ~65% phytoplankton and 35% terrestrial POM. The mean annual export fluxes were 40.6 tPOC/year and 2.45 tPN/year over the studied period, with half of it originating from phytoplankton (53 and 55% for POC and PN, respectively). We observed a clear seasonal pattern in POM composition: phytoplankton predominated from March to October, in relation to high primary production, while terrestrial contributions were the highest from November to February, driven by greater autumn-winter hydrodynamics. Our study illustrate the interest of such an approach to quantify POM composition in aquatic system and estimate source fluxes, and provide fundamental results for estimating seasonal baselines in food webs, establishing biogeochemical budgets, and quantifying POM exports to estuarine and marine environments. Applying this methodology across a broad spectrum of aquatic systems should enhance our understanding of biogeochemical processes and organic matter transformation along the land-ocean continuum and illustrates the contribution of these ecosystems to global biogeochemical cycles.

Mitigation of ammonia volatilization from organic and inorganic nitrogen sources applied to soil using water hyacinth biochars

The recent global population explosion has increased people’s food demand. To meet this demand, huge amounts of nitrogen (N) fertilizer have been applied in the worldwide. However, ammonia (NH3) volatilization is one of the primary factors of N loss from soil after N application causing decrease crop N utilization efficiency and productivity. Incubation experiments were conducted on an acidic clayey soil with two different N sources (urea and anaerobic digestion effluent; ADE), two differently-produced biochars, and three biochar application rates (0%, 0.25%, and 1.0% w/w). Ammonia volatilization was lower from urea (14.0–23.5 mg N kg−1) and ADE (11.3–21.0 mg N kg−1) with biochar application than those without biochar (40.1 and 26.2 mg N kg−1 from urea and ADE alone, respectively). Biochar application significantly mitigated volatilization and reduction percentages for urea and ADE were 40%–64% and 18%–55%, respectively. 1.0% biochar application mitigated volatilization significantly compared to 0.25% application regardless of N source and biochar types. Possible mechanism for volatilization mitigation for urea and ADE were increased N immobilization by soil microorganisms and accelerated net nitrification rate due to increased soil nitrifying bacteria, respectively. Overall, our results clarified different mechanisms for N volatilization mitigation from different (inorganic vs. organic) N sources with biochar application.

Effect of Organic and Inorganic Sources of Nitrogen on Growth Parameter and Yield Attribute of Wheat in Inceptisolof Eastern Uttar Pradesh, India

The insufficient availability and escalating expenses of fertilizers present significant constraints to crop production, exacerbating the growing disparity between nutrient demand and supply. The overreliance on agrochemicals like synthetic fertilizers, pesticides, herbicides, and fungicides not only endangers human health but also degrades soil quality and environmental integrity. This study examined the impact of applying recommended doses of fertilisers (RDF) in conjunction with organic amendments in a pot experiment carried out throughout the 2020–2021 wheat growing season. Significant improvements were found for the following wheat plant attributes: straw and grain yields; height; greenness index; spike count; spike length; and weight of 1000 seeds. The treatment utilizing 50% RDF and 50% poultry manure (T5) demonstrated the most pronounced improvements across these parameters. These findings highlight integrated nutrient management's crucial importance as a modern agricultural requirement.

Recent progress and potential future directions to enhance biological nitrogen fixation in faba bean (Vicia faba L.).

The necessity for sustainable agricultural practices has propelled a renewed interest in legumes such as faba bean (Vicia faba L.) as agents to help deliver increased diversity to cropped systems and provide an organic source of nitrogen (N). However, the increased cultivation of faba beans has proven recalcitrant worldwide as a result of low yields. So, it is hoped that increased and more stable yields would improve the commercial success of the crop and so the likelihood of cultivation. Enhancing biological N fixation (BNF) in faba beans holds promise not only to enhance and stabilize yields but also to increase residual N available to subsequent cereal crops grown on the same field. In this review, we cover recent progress in enhancing BNF in faba beans. Specifically, rhizobial inoculation and the optimization of fertilizer input and cropping systems have received the greatest attention in the literature. We also suggest directions for future research on the subject. In the short term, modification of crop management practices such as fertilizer and biochar input may offer the benefits of enhanced BNF. In the long term, natural variation in rhizobial strains and faba bean genotypes can be harnessed. Strategies must be optimized on a local scale to realize the greatest benefits. Future research must measure the most useful parameters and consider the economic cost of strategies alongside the advantages of enhanced BNF.

Alkaliphilic and thermostable lipase production by leaf litter fungus Leptosphaerulina trifolii A SMR-2011.

Fungi that inhabit fire-prone forests have to be adapted to harsh conditions and fungi affiliated to Ascomycota recovered from foliar litter samples were used for bioprospecting of molecules such as enzymes. Agni's fungi isolated from leaf litter, whose spores are capable of tolerating 110 oC were screened for thermostable lipases. One of the isolates, Leptosphaerulina trifolii A SMR-2011exhibited high positive lipase activity than other isolates while screening through agar plate assay using Tween 20 in the medium. Maximum lipase activity (173.2 U/mg) of L. trifolii was observed at six days of inoculation and decreased thereafter. Among different oils used, the maximum lipase activity was attained by soybean oil (940.1 U/mg) followed by sunflower oil (917.1 U/mg), and then by mustard oil (884.8 U/mg), showing its specificity towards unsaturated fatty acids. Among the various organic nitrogen sources tested, soybean meal showed maximum lipase activity (985.4 U/mg). The partially purified enzyme was active over a wide range of pH from 8 to 12 with a pH optimum of 11.0 (728.1 U/mg) and a temperature range of 60-80 oC with an optimal temperature of 70 oC (779.1 U/mg). The results showed that lipase produced by L. trifolii is alkali stable and retained 85% of its activity at pH 11.0. This enzyme also showed high thermal stability retaining more than 50% of activity when incubated at 60 oC to 90°C for 2h. The ions Ca2+ and Mn2+ induced the lipase activity, while Cu2+ and Zn2+ ions lowered the activity compared to control. These results suggests that the leaf litter fungus L. trifolii serves as a potential source for the production of alkali-tolerant and thermostable lipase.

Exploiting the roles of nitrogen sources for HEA increment in Cordyceps cicadae.

Cordyceps cicadae, as a new food ingredient, is a valuable edible and medicinal fungi. However, its resources are severely depleted due to environmental limitations and excessive harvesting practices. N6-(2-hydroxyethyl) adenosine (HEA), as an important product of Cordyceps cicadae, has the potential to be used in medical industry due to its diverse disease curing potential. However, the disclosure of HEA synthesis still severely limited its application until now. In this study, the kinetic curves for adenosine and HEA under shaker fermentation were explored. The kinetics of HEA and adenosine production exhibited a competitive pattern, implicating a possibility of sharing a same step during their synthesis. Due to HEA as a derivative of nitrogen metabolism, the effect of different nitrogen sources (peptone, yeast extract, ammonium sulfate, diammonium oxalate monohydrate, ammonium citrate dibasic, and ammonium citrate tribasic) on HEA production in Cordyceps cicadae strain AH 10-4 had been explored under different incubation conditions (shaker fermentation, stationary fermentation, and submerged fermentation). Our results indicated that the complex organic nitrogen sources were found to improve the accumulation of HEA content under shaker fermentation. In contrast, the optimal nitrogen source for the accumulation of HEA under stationary fermentation and submerged fermentation was ammonium citrate tribasic. But submerged fermentation obviously shortened the incubation time and had a comparable capacity of HEA accumulation by 2.578 mg/g compared with stationary fermentation of 2.535 mg/g, implicating a possibility of scaled-up production of HEA in industry by submerged fermentation. Based on the dramatic HEA production by ammonium sulfate as nitrogen resources between stationary and shaker fermentations, alanine, aspartate and glutamate as well as arginine metabolic pathway were related to the production of HEA by comparative transcriptome. Further investigation indicated that glutamic acid, which is an analog of Asp, showed an optimum production of HEA in comparison with other amino acids.

Association between organic nitrogen substrates and the optical purity of d-lactic acid during the fermentation by Sporolactobacillus terrae SBT-1

The development of biotechnological lactic acid production has attracted attention to the potential production of an optically pure isomer of lactic acid, although the relationship between fermentation and the biosynthesis of highly optically pure d-lactic acid remains poorly understood. Sporolactobacillus terrae SBT-1 is an excellent d-lactic acid producer that depends on cultivation conditions. Herein, three enzymes responsible for synthesizing optically pure d-lactic acid, including d-lactate dehydrogenase (D-LDH; encoded by ldhDs), l-lactate dehydrogenase (L-LDH; encoded by ldhLs), and lactate racemase (Lar; encoded by larA), were quantified under different organic nitrogen sources and concentration to study the relationship between fermentation conditions and synthesis pathway of optically pure lactic acid. Different organic nitrogen sources and concentrations significantly affected the quantity and quality of d-lactic acid produced by strain SBT-1 as well as the synthetic optically pure lactic acid pathway. Yeast extract is a preferred organic nitrogen source for achieving high catalytic efficiency of d-lactate dehydrogenase and increasing the transcription level of ldhA2, indicating that this enzyme plays a major role in d-lactic acid formation in S. terrae SBT-1. Furthermore, lactate racemization activity could be regulated by the presence of d-lactic acid. The results of this study suggest that specific nutrient requirements are necessary to achieve a stable and highly productive fermentation process for the d-lactic acid of an individual strain.

Complex nitrogen sources from agro-industrial byproducts: impact on production, multi-stress tolerance, virulence, and quality of Beauveria bassiana blastospores.

Biological control through entomopathogenic fungi provides essential ecological services in the integrated management of agricultural pests. In the context of submerged liquid fermentation, the nutritional composition significantly influences the ecological fitness, virulence and quality of these fungi. This study specifically explores the impact of various complex organic nitrogen sources derived from agro-industrial byproducts on the submerged liquid fermentation of Beauveria bassiana, a versatile entomopathogenic fungus known for producing hydrophilic yeast-like blastospores. Notably, manipulating the nitrogen source during submerged cultivation can influence the quality, fitness, and performance of blastospores. This research identifies cottonseed flour as the optimal low-cost nitrogen source, contributing to increased production yields, enhanced multi-stress tolerance, heightened virulence with extended shelf life and long-term bioactivity. These findings deepen our understanding of the critical role of nitrogen compound selection in liquid media formulation, facilitating the production of ecologically fit and virulent blastospores for more effective pest biocontrol programs.

Azolla as a Source of Organic Nitrogen and its Role in Recycling Agricultural Residues and Improving the Quality of Compost

Azolla as a Source of Organic Nitrogen and its Role in Recycling Agricultural Residues and Improving the Quality of Compost

Multiscale effects of radial mixing on mixotrophic microalgal growth and macromolecular synthesis in tubular bubble-column photobioreactors

Extremophillic microalga Chlorella sorokiniana is grown mixotrophically in a 5 L externally-illuminated tubular bubble-column photobioreactor with an external light intensity of 10,500 lx (≈195 μmol-photons.m−2.s−1) at atmospheric (0.04 %), 1 %, 2 %, 3 %, 4 %, and 5 % CO2 concentrations, with acetic acid and urea as the organic carbon and nitrogen sources, respectively, for 6–8 days. Two different setups are employed: one permits axial air flow, and another promotes radial mixing of algal particles, dissolved CO2 and nutrients along with axial sparging. We show that radial mixing increases biomass yields by reducing both photo-limitation and photo-inhibition up to CO2 concentrations <3 %, above which substrate (CO2) inhibition necessitates lower reactor mixing. Radial mixing increases the lipid yield from 1.06 and 1.61 g/L to 1.27 and 1.78 g/L at 2 % and 3 % CO2, respectively, in nitrogen-limited cases, resulting in a 10–20 % increase. We show that complete reactor mixing, facilitated by the dual effects of branched (radial) and axial sparger, is preferred only in the absence of substrate (CO2) inhibition. Subsequently, we stratify the tubular photobioreactor design strategy into two regimes based on CO2 inlet concentrations. In one regime (for inlet CO2 < 3 %), we recommend the deployment of branched sparger to facilitate both radial and axial mixing simultaneously, thereby creating a well-mixed reactor at lower CO2 concentrations. In the other regime (for inlet CO2 > 3 %), axial sparging alone is recommended since complete mixing of CO2 increases substrate inhibition by reducing the reactor pH below 7.5.

Organic amendments increased Chinese milk vetch symbiotic nitrogen fixation by enriching Mesorhizobium in rhizosphere.

Symbiotic nitrogen fixation of Chinese milk vetch (Astragalus sinicus L.) can fix nitrogen from the atmosphere and serve as an organic nitrogen source in agricultural ecosystems. Exogenous organic material application is a common practice of affecting symbiotic nitrogen fixation; however, the results of the regulation activities remain under discussion. Studies on the impact of organic amendments on symbiotic nitrogen fixation have focused on dissolved organic carbon content changes, whereas the impact on dissolved organic carbon composition and the underlying mechanism remain unclear. In situ pot experiments were carried out using soils from a 40-year-old field experiment platform to investigate symbiotic nitrogen fixation rate trends, dissolved organic carbon concentration and component, and diazotroph community structure in roots and in rhizosphere soils following long-term application of different exogenous organic substrates, i.e., green manure, green manure and pig manure, and green manure and rice straw. Remarkable increases in rate were observed in and when compared with that in green manure treatment, with the greatest enhancement observed in the treatment. Moreover, organic amendments, particularly pig manure application, altered diazotroph community composition in rhizosphere soils, therefore increasing the abundance of the host-specific genus Mesorhizobium. Furthermore, organic amendments influence the diazotroph communities through two primary mechanisms. Firstly, the components of dissolved organic carbon promote an increase in available iron, facilitated by the presence of humus substrates. Secondly, the elevated content of dissolved organic carbon and available iron expands the niche breadth of Mesorhizobium within the rhizosphere. Consequently, these alterations result in a modified diazotroph community within the rhizosphere, which in turn influences Mesorhizobium nodulation in the root and symbiotic nitrogen fixation rate. The results of the present study enhance our understanding of the impact of organic amendments on symbiotic nitrogen fixation and the underlying mechanism, highlighting the key role of dissolved organic carbon composition on diazotroph community composition in the rhizosphere.

Production of arabitol from glycerol by immobilized cells of Wickerhamomyces anomalus WC 1501.

Polyalcohols such as arabitol are among the main targets of biorefineries aiming to upcycle wastes and cheap substrates. In previous works Wickerhamomyces anomalus WC 1501 emerged as an excellent arabitol producer utilizing glycerol. Arabitol production by this strain is not growth associated, therefore, in this study, pre-grown cells were entrapped in calcium alginate beads (AB) and utilized for glycerol transformation to arabitol. Flasks experiments aimed to assess the medium composition (i.e., the concentration of inorganic and organic nitrogen sources and phosphates) and to establish the appropriate carrier-to-medium proportion. In flasks, under the best conditions of ammonium limitation and the carrier:medium ratio of 1:3 (w/v), 82.7g/L glycerol were consumed in 168h, yielding 31.2g/L arabitol, with a conversion of 38% and volumetric productivity of 186mg/mL/h. The process with immobilized cells was transferred to laboratory scale bioreactors with different configurations: stirred tank (STR), packed bed (PBR), fluidized bed (FBR), and airlift (ALR) bioreactors. The STR experienced oxygen limitation due to the need to maintain low stirring to preserve AB integrity and performed worse than flasks. Limitations in diffusion and mass transfer of oxygen and/or nutrients characterized also the PBR and the FBR and were partially relieved only in ALR, where 89.4g/L glycerol were consumed in 168h, yielding 38.1g/L arabitol, with a conversion of 42% and volumetric productivity of 227mg/mL/h. When the ALR was supplied with successive pulses of concentrated glycerol to replenish the glycerol as it was being consumed, 117g/L arabitol were generated in 500h, consuming a total of 285g/L glycerol, with a 41% and 234mg/L/h. The study strongly supports the potential of W. anomalus WC 1501 for efficient glycerol-to-arabitol conversion using immobilized cells. While the yeast shows promise by remaining viable and active for extended periods, further optimization is required, especially regarding mixing and oxygenation. Improving the stability of the immobilization process is also crucial for reusing pre-grown cells in multiple cycles, reducing dead times, biomass production costs, and enhancing the economic feasibility of the process.

A life cycle assessment of early-stage enzyme manufacturing simulations from sustainable feedstocks

Enzyme-catalyzed reactions have relatively small environmental footprints. However, enzyme manufacturing significantly impacts the environment through dependence on traditional feedstocks. With the objective of determining the environmental impacts of enzyme production, the sustainability potential of six cradle-to-gate enzyme manufacturing systems focusing on glucose, sea lettuce, acetate, straw, and phototrophic growth, was thoroughly evaluated. Human and ecosystem toxicity categories dominated the overall impacts. Sea lettuce, straw, or phototrophic growth reduces fermentation-based emissions by 51.0, 63.7, and 79.7%, respectively. Substituting glucose-rich media demonstrated great potential to reduce marine eutrophication, land use, and ozone depletion. Replacing organic nitrogen sources with inorganic ones could further lower these impacts. Location-specific differences in electricity result in a 14% and a 27% reduction in the carbon footprint for operation in Denmark compared to the US and China. Low-impact feedstocks can be competitive if they manage to achieve substrate utilization rates and productivity levels of conventional enzyme production processes.

Efficient production of butyl butyrate from mannitol by engineered Clostridium tyrobutyricum

Butyl butyrate has a wide range of applications as flavors, solvents, and biofuels. Green production of butyl butyrate by microorganism fermentation in an industry level attracts great attention on the basis of bio-refinery. In this study, a dark horse Clostridium tyrobutyricum was employed for de novo production of butyl butyrate. Herein, reinforcement of EMP and butyl butyrate fermentation pathways (e.g., pfkA, pykA, crt-hbd-bcd/etfAB, adhE2 and VAAT) and regulation of carbon distribution towards butyl butyrate (e.g., CAT1 inactivation or overexpression of NADPH-dependent hbd1) were conducted in C. tyrobutyricum (MUV3) to enhance butyl butyrate production. The outstanding strain MV12 generated 1.8-fold butyl butyrate production compared to the initial strain MUV3. Subsequently, organic nitrogen source concentration was optimized, 21.1 g/L and 30.1 g/L butyl butyrate were obtained in 37 °C and 25 °C, respectively. Ultimately, fed-batch with two-stage temperature shift fermentation was performed for high titer and yield of butyl butyrate production (142.6 g/L titer and 0.28 mol/mol yield) from mannitol. Hence this study presented a microbial fermentation platform for butyl butyrate or other value-added chemicals production from renewable resources.

Bioreactor study of the metabolic repertoire and morphology of actinomycete Streptomyces rimosus ATCC 10970 under various initial concentrations of carbon and nitrogen sources

This work aims at investigating the influence of the initial concentrations of carbon (glucose) and organic nitrogen (yeast extract) sources on Streptomyces rimosus ATCC10970 secondary metabolism in the stirred tank bioreactors. Additionally, glucose utilisation, biomass formation, pH, redox potential and dissolved oxygen levels, and the morphological development of S. rimosus pseudomycelium were studied. Eighteen secondary metabolites were detected by mass spectrometry and identified with the use of the authentic standard, or putatively with the use of literature and database of secondary metabolites. Varied initial yeast extract concentration acted much stronger on the formation of secondary metabolites than glucose did. For example, oxytetracycline was not biosynthesised at high yeast extract concentration while the formation of three other metabolites was enhanced under these conditions. In the case of glucose its increasing initial concentration led to higher secondary metabolite levels with the exception of an unnamed angucycline. High initial yeast extract concentration also drastically changed S. rimosus pseudomycelial morphology from the pelleted to the dispersed one. Ultimately, the cultivation media with the varied initial levels of carbon and nitrogen sources were proved to have the marked effect on S. rimosus secondary metabolism and to be the simplest way to either induce or block the formation of the selected secondary metabolites.

The Effect of Nitrogen-Enriched Partially Burnt Paddy Husk on Growth and Yield of Rice (Oryza Sativa)

Paddy husk can be defined as a material that can be easily accessed by small-scale, resource-poor farmers in Sri Lanka. It is being produced in large quantities during the production of milled rice as by-products. Currently, most paddy husk is underutilized or left unused due to their intrinsic properties. Partially burnt paddy husk (PBPH) is an organic material that is prepared by incomplete combustion of paddy husk. The use of PBPH as a soil amendment to fix soil fertility problems has been well recognized. It contains a higher porous structure and an enhanced adsorption capacity, therefore, has the ability to slow down nutrient leaching by capturing nutrients in its porous structure. In the present study, a farmer-friendly, biochar-based organic nitrogen fertilizer was developed by nitrogen-enriching PBPH using nitrogen-rich liquid fertilizers in order to determine its effect on the growth and yield of rice. A pot experiment was conducted at the Rice Research and Development Institute, Bathalagoda based on a Completely Randomized Design (CRD) with eight treatments and five replicates.The treatments included T1 (Gliricidia extract+PBPH), T2 (cow urine+PBPH), T3 (poultry manure+PBPH), T4 (fish tonic+PBPH), T5 (compost tea+PBPH), T6 (urea+PBPH), T7 (PBPH only) and T8 (no treatment). Nitrogen (N) content of raw materials, liquid fertilizers, N-enriched PBPH, and N uptake of plants were determined using Kjeldahl method. Phosphorus (P) and potassium (K) contents of PBPH were determined by using UV-Visible Spectrophotometer and Flame Photometer, respectively. Plant height and greenness (SPAD results) were recorded as growth parameters and finally, number of tillers, panicles, grains per pot and grain weight per pot were obtained as the yield parameters. Significantly, the highest N content was found in urea liquid fertilizer (3.42%) whereas the lowest was found in Gliricidia extract (0.35%). Significantly, the highest N content (1.735%) of N-enriched PBPHs was found in T6 (urea+PBPH), while the lowest (0.058%) was found in T7 (PBPH only). Significantly, the highest N uptakes (1.98% and 1.86%) of rice plants were found in T6 (urea+PBPH) and T4 (fish tonic+PBPH) respectively while the lowest (0.56%) was found in the T8 (no treatment). SPAD readings, the number of panicles, tillers and grain yield, and biomass were significantly increased in all treatments over T7 (PBPH only) and T8 (no treatment) treatments. The overall study concluded that the application of N-enriched PBPH has improved the growth and yield of rice, therefore it can be used as a farmer-friendly, sustainable, and organic nitrogen source in paddy cultivation in Sri Lanka.&#x0D; Keywords: Nitrogen, Partially burnt paddy husk, Nitrogen enrichment